A weight-reducing traditional Chinese medicine composition and a preparation method thereof

Abstract

The application discloses a kind of slimming traditional Chinese medicine composition and preparation method thereof, it is related to slimming traditional Chinese medicine technical field.Slimming traditional Chinese medicine composition includes the following weight parts of Chinese medicinal materials: 30-45 parts of raw astragalus, 10-15 parts of prepared atractylodes rhizome, 15-23 parts of cassia bark, 20-30 parts of dried tangerine or orange peel, 30-45 parts of lotus leaf, 20-30 parts of raw hawthorn, 15-23 parts of kelp, 10-15 parts of turmeric, 10-15 parts of fried Semen Raphani, 10-15 parts of raw safflower stigma.The preparation method includes the following steps: the weight parts of the above-mentioned Chinese medicinal materials are extracted by water decocting.The traditional Chinese medicine composition of the application has the effects of benefiting qi and invigorating the spleen, resolving phlegm and eliminating dampness, and is also compatible with drugs for warming kidney and assisting yang, eliminating phlegm and resolving mass, and has the combined effects of treating both symptoms and root causes, tonifying deficiency and eliminating excess, which can increase energy consumption of the body, reduce fat generation and storage to achieve the effect of weight loss.The traditional Chinese medicine composition of the application is mainly composed of food-medicines, and can be taken for a long time.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine for weight loss, and in particular to a traditional Chinese medicine composition for weight loss and its preparation method. Background Technology

[0002] With rapid economic development, the incidence of obesity is increasing, and obesity has become a serious global public health problem. Adipose tissue is the main metabolic organ regulating the body's energy homeostasis and is closely related to the occurrence and development of obesity. Adipose tissue is mainly divided into three types: white adipose tissue (WAT), brown adipose tissue (BAT), and beige adipose tissue. Brown adipose tissue is morphologically and functionally distinct from white adipose tissue. White adipocytes have large and few lipid droplets, mainly used for energy storage, and their significantly increased content is one of the main characteristics of obese patients. Brown adipocytes have small and numerous lipid droplets, mainly used for heat production, and can consume the stored amount of white adipose tissue. Beige adipocytes exist within white adipocytes and are induced thermogenic cells. When stimulated, they can exhibit the characteristics of brown adipocytes; this process is called "white adipose tissue browning" or "browning."

[0003] Modern research suggests that obesity involves two processes: energy intake and energy expenditure. When intake exceeds expenditure, excess energy is stored in the body as fat. This leads to an increase in the number and size of fat cells, causing fat accumulation, an increased body mass index, and thus obesity. Obesity impairs the body's browning process, but studies have shown that stimulation from factors such as cold, exercise, diet, hormones, or chemical treatments can re-induce white fat browning in obese individuals. Inducing browning to increase energy expenditure is crucial for improving metabolic diseases such as obesity and is currently a hot research topic.

[0004] Currently, the incidence of obesity in my country continues to rise, while the cure rate and treatment methods both need improvement. Lifestyle interventions alone are insufficient to achieve effective and lasting weight loss; while surgical treatment is highly effective, it carries risks; and the variety of weight loss products on the market is limited and has certain limitations. Therefore, the development of safer and more effective weight loss drugs is of great urgency and social significance. This study focuses on experimental research on the traditional Chinese medicine composition of this invention, laying the foundation for the development of safe and effective traditional Chinese medicine weight loss products. Summary of the Invention

[0005] The technical problem this invention aims to solve is to provide a traditional Chinese medicine composition for weight loss and its preparation method. This composition has the effects of invigorating qi and strengthening the spleen, resolving phlegm and eliminating dampness. It also incorporates herbs that warm the kidneys and assist yang, and resolve phlegm and dissipate nodules. The combined effect addresses both the symptoms and the root cause, replenishing deficiencies and transforming excesses. It may achieve weight loss by increasing the body's energy consumption and reducing fat production and storage. Furthermore, many of the components of this composition are food-grade medicinal materials, allowing for prolonged use.

[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is: a weight-loss traditional Chinese medicine composition, comprising the following traditional Chinese medicinal materials in parts by weight: 30-45 parts of raw Astragalus membranaceus, 10-15 parts of processed Atractylodes lancea, 15-23 parts of cinnamon, 20-30 parts of dried tangerine peel, 30-45 parts of lotus leaf, 20-30 parts of raw hawthorn, 15-23 parts of kelp, 10-15 parts of turmeric, 10-15 parts of stir-fried radish seed, and 10-15 parts of raw Typha pollen.

[0007] Preferably, the weight-loss traditional Chinese medicine composition includes the following traditional Chinese medicinal materials in parts by weight: 30 parts of raw Astragalus membranaceus, 10 parts of processed Atractylodes lancea, 15 parts of cinnamon, 20 parts of dried tangerine peel, 30 parts of lotus leaf, 20 parts of raw hawthorn, 15 parts of kelp, 10 parts of turmeric, 10 parts of stir-fried radish seed, and 10 parts of raw Typha pollen.

[0008] The preparation method of the above-mentioned weight-loss traditional Chinese medicine composition includes the following steps: extracting the above-mentioned weight parts of traditional Chinese medicine by decoction with water.

[0009] Preferably, the preparation method of the weight-loss traditional Chinese medicine composition includes the following steps: soaking the above-mentioned parts by weight of traditional Chinese medicine materials in water, extracting by decoction, and concentrating.

[0010] Preferably, the water decoction extraction is performed twice.

[0011] Preferably, the preparation method of the weight-loss traditional Chinese medicine composition includes the following steps: soaking the above-mentioned weight parts of Chinese medicinal materials in 10-15 times the weight of the Chinese medicinal materials in water for 0.5-1 hour, then simmering over high heat until boiling, then simmering over low heat for 30-40 minutes, and then pouring out the first decoction; for the second decoction, add 8-12 times the weight of the Chinese medicinal materials in water and continue simmering, then simmering for 20-30 minutes after boiling, and then pour out the second decoction; combine the two decoctions; heat and concentrate the combined decoction, and then refrigerate it.

[0012] A further preferred method for preparing the weight-loss traditional Chinese medicine composition includes the following steps: soaking the above-mentioned weight parts of Chinese medicinal materials in water equal to 10 times their weight for 1 hour, then simmering over high heat until boiling, then simmering over low heat for 30 minutes and pouring out the first decoction; adding water equal to 8 times the weight of the Chinese medicinal materials to the second decoction and simmering for 30 minutes after boiling, then pouring out the second decoction; combining the two decoctions; heating and concentrating the combined decoction, and then refrigerating it for storage.

[0013] More preferably, the heating and concentration temperature is 80-100℃, and the concentration is carried out to 0.8-3.5g·ml. -1 .

[0014] Traditional Chinese medicine theory holds that obesity is closely related to phlegm and dampness. When the spleen and stomach are not in harmony, the transformation and transportation of food and water are impaired, leading to the accumulation of phlegm and dampness. A long-term diet high in fatty, sweet, and greasy foods, coupled with a lack of exercise, gradually leads to the internal generation of phlegm and dampness, which cannot be metabolized, resulting in obesity. This aligns with modern research showing that a long-term high-fat diet leads to the accumulation of white adipose tissue, while exercise can induce browning of white adipose tissue. The herbal composition of this invention focuses on "tonifying qi and strengthening the spleen" and "resolving phlegm and eliminating dampness" as its core therapeutic principles. The combined effects of these herbs warm the yang, tonify qi, strengthen the spleen, and dry dampness.

[0015] "Spleen deficiency and phlegm-dampness" is the key pathogenesis of obesity. Based on his long-term clinical practice, Master Wang Qi proposed that obesity should be treated according to its different types, and created the additive weight loss method. Using "tonifying qi and strengthening the spleen" and "resolving phlegm and eliminating dampness" as the principles of prescription formulation, he developed Yiqi Jianyun Decoction (derived from a combination of a phlegm-resolving and dampness-eliminating formula and Qingjian Capsules). Through continuous optimization and improvement of the formula in clinical intervention, a relatively fixed traditional Chinese medicine composition of this invention has been formed. Clinical cases show that this formula has good weight loss effects.

[0016] This invention comprises a traditional Chinese medicine composition consisting of raw Astragalus membranaceus, processed Atractylodes lancea, cinnamon, tangerine peel, lotus leaf, raw hawthorn, kelp, turmeric, stir-fried radish seed, and raw Typha pollen. Currently, commonly used weight loss methods often focus on reduction, purging, and diarrhea. However, Academician Wang Qi's treatment of obesity takes the opposite approach, creating a weight loss method centered on tonifying Qi and warming Yang. Obesity itself is closely related to spleen Qi deficiency. Raw Astragalus membranaceus in the formula tonifies Qi and promotes diuresis, strengthening the body's digestive and metabolic functions; processed Atractylodes lancea strengthens the spleen and dries dampness. The spleen and stomach are the foundation of acquired constitution, governing digestion and the ascending of clear Qi. Only when the spleen is functioning properly can the generated Qi, blood, and body fluids be distributed to all organs and tissues. Obesity is closely related to spleen deficiency; treating the spleen is the fundamental principle for treating obesity. The alcohol extract of processed Atractylodes lancea can combat diet-induced obesity. Atractylodes lancea extract has anti-inflammatory and diuretic effects, and many other active ingredients in processed Atractylodes lancea also possess anti-inflammatory properties. Spleen deficiency easily leads to phlegm accumulation, hence the combination with tangerine peel to regulate qi and resolve phlegm, and kelp to dispel phlegm and disperse nodules. Turbid accumulation can also lead to blood stasis, and raw hawthorn can aid digestion, resolve stagnation, and promote blood circulation. The tea polyphenols in cinnamon can break down and burn fat, inhibit fat formation, reduce fat storage, and promote gastrointestinal consumption. Lotus leaf can promote urination and bowel movements, and can also form a fat-isolating membrane on the intestinal wall, preventing fat absorption, breaking down fat, and excreting it from the body. Curcumin in turmeric regulates lipid metabolism, reduces the accumulation of fat in the body, and promotes fat burning. Fried radish seeds have the effect of regulating qi and resolving phlegm, and promoting gastrointestinal motility. Raw cattail pollen promotes blood circulation, removes blood stasis, has a good lipid-lowering effect, and strengthens the spleen and replenishes qi.

[0017] The beneficial effects of adopting the above technical solution are as follows:

[0018] (1) The herbal composition of this invention has the effects of invigorating qi and strengthening the spleen, resolving phlegm and removing dampness. At the same time, it is also combined with herbs that warm the kidneys and assist yang, and resolve phlegm and dissipate nodules. The combined effect of the formula can treat both the symptoms and the root cause, replenish deficiency and resolve excess. It may achieve the effect of weight loss by increasing the body's energy consumption and reducing the production and storage of fat. Moreover, most of the herbs in the herbal composition of this invention are food and medicine of the same origin, and can be taken for a long time.

[0019] (2) The herbal composition of this invention can target adipose tissue, promote the browning of white adipose tissue by regulating the level of browning factor in white adipose tissue, and activate the activity of brown adipose tissue in the body, thereby increasing the expression of UCP1 and promoting the body's energy consumption. The experiments of this invention have verified the formulation idea of ​​the herbal composition of this invention. The experimental results prove that the herbal composition of this invention can indeed effectively reduce the accumulation of white adipose tissue and achieve the purpose of weight loss by promoting the browning of white adipose tissue and increasing the energy consumption of obese rats.

[0020] (3) The herbal composition of this invention can reduce the body weight of obese rats, regulate the levels of blood lipids, inflammatory factors, insulin, free fatty acids, and serum leptin, and significantly improve the morphology of adipose tissue cells and increase the expression of related browning factors in inguinal white adipose tissue. These results indicate that the herbal composition of this invention can resist obesity induced by a high-fat diet. Its mechanism may be through inducing browning of white adipose tissue and increasing the activity of brown adipose tissue, with a particularly significant promoting effect on subcutaneous white adipose tissue. Attached Figure Description

[0021] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments;

[0022] Figure 1 This is a graph showing the change in average body weight (g) of rats in each group during the administration period of this invention;

[0023] Figure 2 This is a bar graph showing the average abdominal circumference of rats in each group after 8 weeks of administration according to the present invention;

[0024] Figure 3 This is a bar graph showing the changes in average food intake of rats in each group during the administration period of this invention;

[0025] Figure 4 This is a bar chart comparing serum insulin levels in rats of different groups after drug intervention according to the present invention;

[0026] Figure 5 This is a diagram showing the morphological changes of rat liver tissue in this invention (HE, ×200);

[0027] Figure 6 This is a diagram showing the morphological changes of subcutaneous white adipose tissue in rats according to the present invention (HE, ×200);

[0028] Figure 7 This is a diagram showing the morphological changes of white adipose tissue in the rat viscera according to the present invention (HE, ×200);

[0029] Figure 8 This is a diagram showing the changes in the morphology of brown adipose tissue in rats according to the present invention (HE, ×200);

[0030] Figure 9 This is an electrophoresis diagram of the PCR products of this invention;

[0031] Figure 10 This is a bar chart comparing the 2-ΔΔCT values ​​of various browning factors within the iWAT of this invention;

[0032] Figure 11 This is a bar chart comparing the 2-ΔΔCT values ​​of various browning factors within the eWAT of this invention;

[0033] Figure 12 The browning factors 2 in the BAT of this invention -△△CT A bar chart comparing values;

[0034] Figure 13 This is a bar chart showing the relative protein expression levels of various browning factors within the iWAT of this invention;

[0035] Figure 14 This is a protein band diagram of each browning factor in the iWAT of this invention;

[0036] Figure 15 This is a line graph showing the weight changes of the experimental and control groups in this invention.

[0037] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group); # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01, ### means P < 0.001; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01; Figure 5-8 This is a scan image from a super-resolution microscopic tissue imaging system. Detailed Implementation

[0038] Example 1

[0039] A weight-loss traditional Chinese medicine composition comprises the following Chinese medicinal materials in parts by weight: 30 parts raw Astragalus membranaceus, 10 parts processed Atractylodes lancea, 15 parts cinnamon, 20 parts dried tangerine peel, 30 parts lotus leaf, 20 parts raw hawthorn, 15 parts kelp, 10 parts turmeric, 10 parts stir-fried radish seed, and 10 parts raw Typha pollen.

[0040] The preparation method includes the following steps: Soak the above-mentioned medicinal materials in 10 times their weight of water for 1 hour, then bring to a boil over high heat. After boiling, reduce to a simmer and continue simmering for 30 minutes. Pour out the first decoction. For the second decoction, add 8 times the weight of hot water to the medicinal materials and continue simmering. After boiling, simmer for 30 minutes, then pour out the second decoction. Combine the two decoctions. Heat and concentrate the combined decoction to 3.2546 g / ml. -1 Store in the refrigerator.

[0041] Example 2

[0042] A weight-loss traditional Chinese medicine composition comprises the following Chinese medicinal materials in parts by weight: 45 parts raw Astragalus membranaceus, 15 parts processed Atractylodes lancea, 23 parts cinnamon, 30 parts dried tangerine peel, 45 parts lotus leaf, 30 parts raw hawthorn, 23 parts kelp, 15 parts turmeric, 15 parts stir-fried radish seed, and 15 parts raw Typha pollen.

[0043] The preparation method includes the following steps: Soak the above-mentioned medicinal materials in 15 times their weight of water for 0.5 hours, then bring to a boil over high heat. After boiling, reduce to a simmer and continue simmering for 40 minutes. Pour out the first decoction. For the second decoction, add 10 times the weight of hot water and continue simmering until boiling. Simmer for 25 minutes, then pour out the second decoction. Combine the two decoctions. Heat and concentrate the combined decoction to 3.2546 g / ml. -1 Store in the refrigerator.

[0044] Example 3

[0045] A weight-loss traditional Chinese medicine composition comprises the following parts by weight of Chinese medicinal materials: 40 parts of raw Astragalus membranaceus, 12 parts of processed Atractylodes lancea, 18 parts of cinnamon, 24 parts of dried tangerine peel, 36 parts of lotus leaf, 24 parts of raw hawthorn, 18 parts of kelp, 12 parts of turmeric, 12 parts of stir-fried radish seed, and 12 parts of raw Typha pollen.

[0046] The preparation method includes the following steps: Soak the above-mentioned medicinal materials in 12 times their weight of water for 0.8 hours, then bring to a boil over high heat. After boiling, reduce to a simmer and continue simmering for 35 minutes. Pour out the first decoction. For the second decoction, add 12 times the weight of hot water and continue simmering until boiling. Simmer for 20 minutes, then pour out the second decoction. Combine the two decoctions. Heat and concentrate the combined decoction to 3.2546 g / ml. -1 Store in the refrigerator.

[0047] Experimental research

[0048] Experiment 1: Establishment of a nutritionally obese rat model

[0049] 1. Materials

[0050] 1.1 Laboratory Animals and Their Care

[0051] Eighty healthy male SD rats, 6 weeks old, weighing 220±20g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed at the animal facility of Beijing University of Chinese Medicine (SPF grade, temperature 22℃, humidity 46%, 12h / 12h light / dark cycle). A high-fat diet (composed of 70% basal diet + 15% lard + 15% sucrose) was purchased from Spiefer (Beijing) Biotechnology Co., Ltd.; a standard maintenance diet was provided by the animal facility of Beijing University of Chinese Medicine.

[0052] 1.2 Experimental Equipment

[0053] Electronic balance, Shanghai Mettler Toledo Instruments Co., Ltd., China; measuring tape.

[0054] 2 methods

[0055] 2.1 Grouping of experimental animals

[0056] All rats were given an acclimatization diet for one week prior to the experiment, during which they had free access to food and water. Rats with no adverse reactions and normal food, water intake, and activity were included in the experiment. After one week of acclimatization, 80 rats were randomly divided into a normal diet group (n=10) and a high-fat diet group (n=70).

[0057] 2.2 Animal Model Preparation

[0058] The normal diet group was fed a normal maintenance diet, while the high-fat diet group was fed a high-fat diet (composed of 70% basal diet + 15% lard + 15% sucrose). The rearing environment was kept at a constant temperature and humidity, and all rats had free access to food and water.

[0059] 2.3 Indicator Testing

[0060] Measure weight and body length weekly and calculate Lee's Index (KPI) using the following formula: LEE'S INDEX=Weight(g)^(1 / 3) x10 3 Body length (cm) .

[0061] 2.4 Statistical Analysis

[0062] Statistical analysis was performed using SPSS 26 statistical software. The data were presented as follows: The data were then subjected to a normality test. For data that conformed to a normal distribution after the test, paired t-tests were used before and after the intervention; for data that were not normally distributed, t' tests were used. A p-value < 0.05 was considered statistically significant.

[0063] 3 Results

[0064] At the start of modeling, the average body weight of the two groups of rats differed by 3.01 g, which was not statistically significant (P > 0.05). At week 10 of modeling, the average body weight of the high-fat diet group exceeded that of the normal diet group by 20.33% (meeting the modeling standard, > 20%), and this difference was highly significant, indicating a successful establishment of a nutritionally ill (HFD) obese rat model.

[0065] Table 1-1 Changes in mean body weight of rats in the two groups during modeling (g, )

[0066]

[0067] Note: * indicates that compared with the normal feed group, * means P < 0.05; ** means P < 0.01; *** means P < 0.001.

[0068] At the beginning of the modeling process, there was no significant difference in Lee's index between the two groups of rats (P > 0.05), but a highly significant difference appeared between the two groups at week 10 (P < 0.01). The specific values ​​are shown in the table below:

[0069] Table 1-2 Comparison of mean Lee's index between the two groups of rats before and after modeling.

[0070]

[0071] Note: * indicates that compared with the normal feed group, ** means P < 0.01.

[0072] 4 Discussion

[0073] In traditional animal models of obesity, rats of the same breed are typically induced directly using a high-fat diet. However, there are no reports on whether there are differences between the commonly used SD and Wistar rat breeds in China. Studies comparing the two breeds in terms of body weight, body length, Lee's index, food intake, food utilization, body fat mass and fat / body ratio, and blood biochemical indicators found no statistically significant differences between the two model groups, indicating that either SD or Wistar rats can be used to establish an obesity model.

[0074] Establishing an obese rat model using a high-fat diet is a relatively mature method. Currently, there is no unified standard for judging the success of an obese animal model, both domestically and internationally. Most methods use indicators such as the percentage of body mass exceeding the average body mass of the control group, body mass gain, fat / body mass ratio, and Lee's index to determine model success. In this experiment, at the beginning of modeling, there was no difference in average body weight and Lee's index between the normal diet group and the high-fat diet group. However, significant differences appeared between the two groups at the end of modeling, indicating that the obese rat model was successfully constructed in this experiment.

[0075] As research in the field of obesity continues to advance, an increasing number of studies tend to exclude obesity-resistant rats from obesity-susceptible rats during the establishment of obesity models, thereby making the obesity models more representative. This experiment also excluded the 20 rats with the lowest body weight.

[0076] Experiment 2: Evaluation of the weight loss and lipid-lowering effects of the traditional Chinese medicine composition of this invention.

[0077] 1. Materials

[0078] 1.1 Experimental Reagents

[0079] Chinese medicinal herbs (processed slices) were purchased from Beijing Tongrentang Xinyuedu Store; Orlistat capsules were purchased from Beijing Gaoyuan Baikang Pharmacy; Anhydrous ethanol, anhydrous glucose powder, and physiological saline were all purchased from Beijing Bainuowei Biotechnology Co., Ltd.; Roche blood glucose meter and Roche blood glucose test strips were both purchased from Meikang Instrument Equipment (Beijing) Co., Ltd.; PBS buffer was purchased from Wuhan Sewell Biotechnology Co., Ltd.; Sodium pentobarbital solution was purchased from the warehouse of Beijing University of Chinese Medicine; Paraformaldehyde fixative was purchased from Beijing Lanbolide Trading Co., Ltd.; Xylene, paraffin, hematoxylin, and eosin were all purchased from Beijing Honghu United Chemical Products Co., Ltd.

[0080] 1.2 Experimental Equipment and Instruments

[0081] Syringes of various sizes, intravenous blood collection needles, beakers, filter paper, medical scissors, medical forceps, centrifuge tubes, pipettes; 5415 high-speed centrifuge, Eppendorf, Germany; ultramicrotome, super-resolution microtissue imaging system, Leica, USA.

[0082] 2 methods

[0083] 2.1 Preparation of the weight-loss traditional Chinese medicine composition of the present invention

[0084] Same as Example 1.

[0085] 2.2 Experimental Grouping

[0086] After successful model establishment, all rats were switched to a normal maintenance diet. Rats in the high-fat diet group were ranked from largest to smallest body weight, and the bottom 20 rats were removed. The remaining 50 rats were randomly divided into 5 groups using a random number table: the model group (M), the high-dose group of the weight-loss traditional Chinese medicine composition of this invention (G), the medium-dose group of the weight-loss traditional Chinese medicine composition of this invention (Z), the low-dose group of the weight-loss traditional Chinese medicine composition of this invention (D), and the positive group (Y). The normal diet group was designated as the blank group (K), for a total of 6 groups. The high-dose group (G) had a dosage of 32.546 g / kg. -1 Medium dose group: 16.273 g·kg -1(Z), Low-dose group 8.135 g·kg -1 The drug concentration is achieved by directly boiling out a high dose, which is then diluted into medium and low doses.

[0087] 2.3 Administration method

[0088] (1) Administration method: Administer by gavage once a day for a total of 8 weeks.

[0089] (2) Dosage volume: 1 ml / 100 g body weight.

[0090] (3) Dosage: The three Chinese medicine groups and the positive drug group were converted according to the human-mouse equivalent dose; the blank group and the model group were given an equal amount of physiological saline by gavage.

[0091] 2.4 Sample Preparation and Index Detection

[0092] During the administration period, the rats' body weight, body length (from nose tip to anus), abdominal circumference, and food intake were measured weekly.

[0093] 2.4.1 Preparation and Detection of Rat Serum

[0094] After the drug intervention, the rats were fasted for 12 hours and then anesthetized by intraperitoneal injection of 1% sodium pentobarbital solution. The abdominal cavity was then cut open, and blood was collected from the abdominal aorta. The collected plasma was allowed to stand for 4 hours, then centrifuged at 1500 rpm for 10 minutes at 4°C. The supernatant serum was transferred to a clean 2 ml centrifuge tube using a pipette and stored at -80°C until analysis.

[0095] The levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in serum were detected using a fully automated biochemical analyzer.

[0096] The levels of free fatty acids (FFA), insulin, C-reactive protein (CRP), tumor necrosis factor (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-32 (IL-32) in serum samples were detected using the ELISA kit method according to the kit instructions.

[0097] 2.4.2 Preparation of Liver / Adipose Tissue Sections

[0098] (1) After blood was collected from the abdominal aorta, all of the rat liver, scapular brown fat, inguinal white fat, mesenteric white fat, perirenal white fat and periepididymal white fat were removed, weighed and recorded. After weighing, the liver, brown fat, inguinal white fat and periepididymal white fat were cut into 1cm*1cm cubes with a blade and fixed in 4% paraformaldehyde fixative.

[0099] (2) Remove formaldehyde solution, rinse tissue with deionized water to remove residual paraformaldehyde, and transfer tissue into 50% alcohol solution for rinsing.

[0100] (3) The rinsed tissue was transferred into gradient alcohol solutions with concentrations of 70%, 80%, and 90% and dehydrated for 1 hour at room temperature.

[0101] (4) Continue to transfer the tissue into 100% alcohol solution I and dehydrate for 1 hour at room temperature;

[0102] (5) Continue to transfer the tissue into 100% alcohol solution II and dehydrate for 1 hour at room temperature;

[0103] (6) Transfer the dehydrated tissue into xylene solution I and leave it in the solution for 30 minutes at room temperature;

[0104] (7) Transfer the treated tissue into xylene solution II, allow it to clear for 30 minutes, then allow it to sit at room temperature;

[0105] (8) After the tissue has become transparent, it is placed into pre-melted paraffin liquid I and kept at a constant temperature of 60°C for 2 hours. Do not exceed 62°C.

[0106] (9) Continue to transfer the tissue into the pre-melted paraffin liquid II, keep the temperature constant at 60°C, and soak for 2 hours;

[0107] (10) Place the soaked tissue in a paraffin embedding container, and after the paraffin cools, place it in a 4°C refrigerator overnight;

[0108] (11) Fix the embedded tissue onto a microtome and slice it;

[0109] (12) Spreading and scooping the slide. Transfer the relatively intact paraffin section to a 45°C water bath. After the surface is flat and wrinkle-free, scoop it up with a glass slide coated with protein glycerin.

[0110] (13) Place the paraffin slices in an oven at 45°C for 48 hours to ensure that the paraffin slices and glass slides are firmly bonded, flat, and free of air bubbles.

[0111] 2.4.3 Specific steps for HE staining of paraffin sections

[0112] (1) Immerse the paraffin slices in xylene I for dewaxing for 5 minutes;

[0113] (2) Transfer the dewaxed slices into xylene II and soak for 5 minutes;

[0114] (3) The slices were immersed in alcohol solutions of 100%, 95%, 90%, 80% and 75% in descending order of concentration, with each concentration being immersed for 3 minutes;

[0115] (4) Immerse the soaked slices in distilled water and rinse for 5 minutes;

[0116] (5) Transfer the rinsed slices to the hematoxylin staining solution and react for 5 minutes;

[0117] (6) Transfer the section to eosin staining solution and react for 5 minutes;

[0118] (7) The slices were quickly rinsed in alcohol solutions of 75%, 80%, 90%, and 95% concentrations, respectively, from low to high concentration.

[0119] (8) Immerse the rinsed slices in a 100% alcohol solution for 1 minute;

[0120] (9) Transfer the slices into xylene I and continue soaking for 5 minutes;

[0121] (10) Transfer the slices into xylene II and continue soaking for 5 minutes;

[0122] (11) After removing the sections, place them in a fume hood to dry, and then seal them with neutral resin.

[0123] (12) Observe and photograph under an optical microscope after 48 hours.

[0124] 2.5 Statistical Analysis

[0125] Statistical analysis was performed using SPSS 26 statistical software. The data were presented as follows: The data were presented as follows. Normality tests were performed on each group of data. For groups conforming to a normal distribution, paired t-tests were used before and after the intervention; for non-normally distributed data, t' tests were used; independent samples t-tests were used to compare data from two groups that conformed to a normal distribution; one-way ANOVA was used for comparisons among multiple groups, with LSD and Tamhane's T2 methods used for homogeneous or unequal variances, respectively; if the data did not conform to a normal distribution, the rank-sum test for multiple independent samples was used. A p-value < 0.05 was considered statistically significant; graphs were generated using Graphpad Prism 8.0 statistical software.

[0126] 3 Results

[0127] 3.1 Comparison of morphological indices among rats in each group

[0128] 3.1.1 Body weight

[0129] From Table 2-1, Figure 1It was found that after 8 weeks of intervention, the average weight of rats in the high-dose group was significantly lower than that in the model group, with a statistically significant difference (P < 0.01); the average weight of rats in the medium-dose group was also significantly lower than that in the model group, with a similarly statistically significant difference (P < 0.05); the average weight of rats in the low-dose group was also somewhat lower than that in the model group, but the difference was not statistically significant (P > 0.05); the average weight of rats in the positive control group was also significantly lower than that in the model group, but the difference was not statistically significant (P > 0.05). These results indicate that the herbal composition of this invention has a good weight-loss effect on nutritionally obese rats, especially with high and medium doses, which are slightly better than the positive control drug orlistat.

[0130] Table 2-1 Changes in mean body weight of rats in different groups during drug administration (g, )

[0131]

[0132] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, ## means P < 0.01, ### means P < 0.001; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01.

[0133] 3.1.2 Lee's Index

[0134] As shown in Table 2-2, after 8 weeks of drug intervention, the Lee's index of rats in each treatment group was slightly lower than that in the model group. The high-dose group and the positive control group showed better results than the medium- and low-dose groups, but the Lee's index of the high-dose group was slightly higher than that of the positive control group. There were no statistically significant differences among the four groups (P > 0.05). The Lee's index of all three traditional Chinese medicine groups was higher than that of the positive control group.

[0135] Table 2-2 Mean Lee's Index of Rats in Each Group After 8 Weeks of Drug Intervention

[0136]

[0137] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group).

[0138] 3.1.3 Waist circumference

[0139] Table 2-3 shows the changes in the average abdominal circumference of rats in each group during drug intervention. Figure 2As can be seen, after 8 weeks of intervention, the abdominal circumference of all three TCM groups and the positive group decreased to varying degrees compared with the model group, with the high-dose group and the positive group showing a smaller decrease than the medium- and low-dose groups. The average abdominal circumference of the positive group and the high-dose group was almost identical in value, but the difference between the positive group and the model group was statistically significant (P < 0.05), while there was no statistically significant difference in the three TCM groups (P > 0.05).

[0140] Table 2-3 Changes in mean abdominal circumference of rats in each group during drug administration (cm, )

[0141]

[0142] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01, ### means P < 0.001; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01.

[0143] Bar graphs showing the mean abdominal circumference of rats in each group after 8 weeks of drug administration are shown below. Figure 2 As shown. Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, ## means P < 0.01; * indicates that compared with the model group, * means P < 0.05.

[0144] 3.1.4 Food intake

[0145] Depend on Figure 3 It can be seen that the average food intake of rats in each group did not show a significant pattern during the first four weeks of drug intervention. However, starting from the fifth week, a certain pattern emerged: the food intake of rats in the blank control group and the model group was almost consistent, while the average food intake of rats in the three groups using the herbal composition of this invention and the positive control group was lower than that of the model group, with the high-dose group < medium-dose group < low-dose group < positive control group. This indicates that both the herbal composition of this invention and orlistat inhibited the appetite of obese rats to some extent, with the herbal composition of this invention exhibiting a higher degree of inhibition, and the greater the dose, the greater the impact on food intake.

[0146] Table 2-4 Changes in average food intake (g) of rats in each group during drug administration.

[0147]

[0148] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group).

[0149] Bar graph showing the changes in mean food intake of rats in each group during the drug administration period, see [link to bar graph]. Figure 3 As shown. Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group).

[0150] 3.1.5 Liver and adipose tissue weight

[0151] White adipose tissue is divided into two types: subcutaneous white adipose tissue and visceral white adipose tissue. Subcutaneous white adipose tissue includes mesenteric white adipose tissue and inguinal white adipose tissue, while visceral white adipose tissue includes perirenal white adipose tissue and periepididymal white adipose tissue. After 8 weeks of drug intervention, the liver and various adipose tissues were removed, weighed, and analyzed to obtain the following data.

[0152] Based on the data in Tables 2-5, the weight of various adipose tissues in the model group was significantly higher than that in the control group. Statistically significant differences were observed in inguinal white fat (P < 0.01), total subcutaneous white fat (P < 0.05), perirenal white fat (P < 0.01), periepididymal white fat (P < 0.01), and total visceral white fat (P < 0.01) between groups. No statistically significant differences were found in mesenteric white fat and brown fat between groups (P > 0.05). In the high-dose group, the weight of both subcutaneous and visceral white fat decreased compared to the model group. The weight of subcutaneous white adipose tissue showed a highly significant difference (P < 0.01), visceral white adipose tissue showed a significant difference (P < 0.05), and the weight of brown adipose tissue also showed a significant difference (P < 0.05). The weights of subcutaneous, visceral white adipose tissue, and brown adipose tissue in the medium and low dose groups were also significantly lower than those in the model group, but the differences were not statistically significant (P > 0.05). In the positive control group, the weight of subcutaneous white adipose tissue was significantly lower than that in the model group (P < 0.05), and the weights of visceral white adipose tissue and brown adipose tissue also decreased, but the differences were not statistically significant (P > 0.05). The effect of the positive control group in reducing fat accumulation was far less than that in the high-dose group, comparable to that in the medium-dose group, but better than that in the low-dose group.

[0153] Table 2-5 Mean mass of various tissues in rats of each group after 8 weeks of drug intervention (g, )

[0154]

[0155] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01.

[0156] 3.2 Comparison of serum markers in rats of different groups after drug administration

[0157] 3.2.1 Four blood lipid tests

[0158] Table 2-6 compares the four lipid parameters of rats in each group after drug intervention. After 8 weeks of drug intervention, compared with the blank group, the levels of TG, TC, and LDL-C in the model group increased, while the level of HDL-C decreased, but the differences were not statistically significant (P>0.05). Compared with the model group, the levels of TG, TC, and LDL-C in the serum of rats in the high-dose group decreased, while the level of HDL-C increased, and the four indicators showed no statistically significant differences (P>0.05). Compared with the model group, the levels of TG and LDL-C in the serum of rats in the medium-dose group decreased, while the level of HDL-C increased, with statistically significant differences in TG and HDL-C between groups (P<0.05). Compared with the model group, the levels of TG, TC, and LDL-C in the low-dose group decreased, while the level of HDL-C increased, and the four indicators also showed no statistically significant differences (P>0.05). The levels of TG, TC, and LDL-C in the positive group were lower than those in the model group, with a highly significant difference in LDL-C content (P<0.01), while the level of HDL-C was almost the same as that in the model group. In regulating the four lipid profiles, the high-dose group was more effective than the positive control group, while the positive control group was slightly more effective than the medium- and low-dose groups.

[0159] Table 2-6 Comparison of four lipid parameters in rats of different groups after drug intervention (mmol / L, )

[0160]

[0161] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01.

[0162] 3.2.2 Inflammatory Factors

[0163] The results in Table 2-7 show that the serum levels of TNFα, CRP, IL-6, and IL-10 in the model group rats were significantly higher than those in the model group (P < 0.001, P < 0.001, P < 0.01, P < 0.001, respectively), indicating that the model group rats exhibited a significant inflammatory state. After drug intervention, the inflammatory state of rats in the high-dose group improved significantly. The serum levels of TNFα, CRP, IL-6, and IL-10 were significantly lower than those in the model group, and all were statistically significant (P < 0.001, P < 0.01, P < 0.05, P < 0.01, respectively), and their serum levels were even close to those in the blank group. Although the effects of the medium- and low-dose groups were not as good as those of the high-dose group, the levels of each inflammatory factor were also lower than those in the model group. The levels of CRP and IL-10 in the medium-dose group were significantly different (P < 0.01, P < 0.05), and the CRP level in the low-dose group was statistically different (P < 0.05). The serum levels of TNFα, CRP, IL-6, and IL-10 in the positive group were significantly lower than those in the model group. Among them, the difference between CRP and IL-10 was statistically significant (P < 0.01), while the difference between TNFα and IL-6 was not statistically significant (P > 0.05). Of the four treatment groups, the high-dose group was the most effective at reducing inflammatory factor levels, followed by the positive control drug, while the medium and low doses were not as effective as the positive control drug.

[0164] Table 2-7 Comparison of inflammatory factors in rats of different groups after drug intervention

[0165]

[0166] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01.

[0167] 3.2.3 Serum Insulin

[0168] Depend on Figure 4 It was found that the serum insulin level in the model group was higher than that in the control group, but the difference between the groups was not statistically significant (P>0.05). The three traditional Chinese medicine groups all showed a decrease in insulin level compared to the model group, returning to the control group level, but the difference among the three groups was not statistically significant (P>0.05). The serum insulin level in the positive control group was significantly lower than that in the model group, and the difference between the groups was statistically significant (P<0.05). The positive control group showed a more significant effect in reducing serum insulin levels in obese rats than the three traditional Chinese medicine groups.

[0169] Table 2-8 Comparison of serum insulin levels in rats of different groups after drug intervention

[0170]

[0171] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). * indicates that compared with the model group, * means P < 0.05.

[0172] Bar graph comparing serum insulin levels in different groups of rats after drug intervention, see [image / image]. Figure 4 As shown. Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). * indicates that compared with the model group, * means P < 0.05.

[0173] 3.2.4 Free fatty acids

[0174] In this experiment, the concentration of free fatty acids in the model group rats was slightly increased compared to the control group, but the difference was not significant and was not statistically significant (P>0.05). Compared to the model group, the free fatty acid content in the high, medium, and low-dose herbal medicine groups was significantly decreased, and the differences were all statistically significant (P<0.01, P<0.01, P<0.05). Furthermore, the serum free fatty acid content in all three herbal medicine groups was significantly lower than that in the control group, and the differences were also statistically significant (P<0.01, P<0.01, P<0.05). The free fatty acid content in the positive control group rats was also lower than that in the model group, but the difference was not statistically significant (P>0.05). The herbal medicine composition of this invention has a particularly significant effect on regulating serum free fatty acids; the free fatty acid levels in all three herbal medicine groups were significantly lower than those in the positive control group.

[0175] Table 2-9 Comparison of free fatty acid content in rats of different groups after drug intervention

[0176]

[0177] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01; * indicates that compared with the model group, * means P < 0.05, ** means P < 0.01; & indicates that compared with the positive group, & means P < 0.05.

[0178] 3.2.5 Serum leptin

[0179] The serum leptin levels in the model group rats were significantly higher than those in the control group, with a highly significant difference between the two groups (P < 0.001). After 8 weeks of drug intervention, compared with the model group, the leptin levels in the high-dose, medium-dose, and positive control groups were significantly decreased (P < 0.001, P < 0.01, P < 0.001). Although there was no significant difference between the low-dose group and the model group (P > 0.05), the leptin levels in the low-dose group also decreased to some extent. Both the herbal composition of this invention and orlistat can significantly reduce serum leptin levels in obese rats, with the effect being: high-dose group > positive control group > medium-dose group > low-dose group.

[0180] Table 2-10 Comparison of serum leptin levels in rats of different groups after drug intervention

[0181]

[0182] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, ### means P < 0.001; * indicates that compared with the model group, ** means P < 0.01, and *** means P < 0.001.

[0183] 3.3 Comparison of tissue morphology among rats in different groups after drug administration

[0184] 3.3.1 Morphological changes in the liver

[0185] Depend on Figure 5 The changes in rat liver tissue morphology (HE, ×200) show that the cell morphology and lobular structure of the liver tissue in the blank group were normal, with no diffuse inflammatory cell infiltration, no fat vacuoles, and no lesions observed. The liver tissue of the model group was filled with fat vacuoles. Compared with the model group, the liver tissue of the high, medium, and low dose groups of the herbal composition of this invention, as well as the positive group, showed only a small number of fat vacuoles. The high-dose group had the fewest vacuoles and the smallest size; the medium-dose group had slightly larger vacuoles than the high-dose group, but the number was not large; the low-dose group also showed improvement in the number and volume of vacuoles compared to the model group; the positive group also showed a significant improvement in the number and volume of vacuoles compared to the model group. The number of fat vacuoles in the liver of the positive group rats was comparable to that of the medium-dose group, more than the high-dose group but less than the low-dose group.

[0186] 3.3.2 Morphological changes in adipose tissue

[0187] (1) Morphological changes of iWAT

[0188] Figure 6Image showing changes in the morphology of subcutaneous white adipose tissue in rats (HE, ×200). Electron microscopy shows that the iWAT cells in the control group rats are of normal volume and uniform size; while compared with the control group rats, the iWAT cells in the model group rats are significantly larger in volume and have different shapes and sizes. Under the same area, the number of adipocytes in the model group is less than that in the control group.

[0189] Compared to the model group, the iWAT cells in the high-dose and medium-dose groups were significantly smaller and more uniform in morphology, even comparable to the control group. The number of adipocytes in the same area was also greater than in the model group. The cell volume in the low-dose group was slightly smaller than that in the model group, but the difference was not significant. The cell size in the positive control group was also significantly reduced, and the number of adipocytes was the highest in the same area. The positive control drug is more effective than the herbal composition of this invention in improving the morphology of iWAT adipocytes.

[0190] (2) Morphological changes of eWAT

[0191] Figure 7 The image shows changes in the morphology of white adipose tissue in rat viscera (HE, ×200). Electron microscopy revealed that the eWAT cells in the control group were of normal volume and relatively uniform size. Compared to the control group, the eWAT cell volume in the model group was significantly increased, and the number of adipocytes in the model group was less than that in the control group for the same area. Compared to the model group, the eWAT cells in the high-dose group were significantly smaller and more uniform in morphology, and the number of adipocytes in the same area was also greater than that in the model group. The cell volumes in the medium- and low-dose groups and the positive control group were also smaller than those in the model group, and the cell sizes in these three groups were almost identical. The herbal composition of this invention has a regulatory effect on white adipocytes in the viscera of obese rats comparable to that of orlistat.

[0192] (3) Morphological changes of BAT

[0193] Depend on Figure 8 The image showing changes in the morphology of brown adipose tissue in rats (HE, ×200) reveals that, compared to the control group, the BAT cells in the model group were larger and less uniform in size. Compared to the model group, the high-dose group showed smaller and more uniform BAT cells. The medium-dose and positive control groups also showed smaller and more uniform cell sizes than the model group. The low-dose group also showed a decrease in cell volume compared to the model group, and the cell morphology became more uniform, but the difference was not significant. The high-dose group had the greatest effect on BAT morphology changes, the medium-dose group showed similar results to the positive control group, while the low-dose group was slightly less effective than the first three groups.

[0194] 4 Discussion

[0195] Overweight / obesity is closely related to dyslipidemia. Triglycerides (TG) are important molecules for the body to effectively store excess energy. Obese patients often exhibit hypertriglyceridemia, the mechanism of which is related to insulin resistance, including increased flux of adipose tissue-derived free fatty acids (FFA), leading to increased hepatic triglyceride synthesis and secretion. Total cholesterol (TC) is a steroid hormone, a structural component of cell membranes in all animals, and a precursor to vitamin D and bile acids. Obesity can reduce the uptake and excretion of TC by the liver and fat, resulting in decreased plasma TC clearance and elevated TC levels. The outer layer of LDL-C is composed of phospholipids, free cholesterol, and apolipoprotein B (ApoB), which can carry hydrophobic cholesterol through the blood. Elevated plasma LDL-C levels are usually associated with atherosclerosis. High-density lipoprotein cholesterol (HDL-C) is a complex containing proteins, cholesterol, and phospholipids, mainly synthesized in the liver and small intestine. Its main function is to transport cholesterol to the liver for breakdown and excretion. Obesity can increase TG, TC, and LDL-C levels, and decrease HDL-C levels. The results of this experiment show that although there is no significant difference between the traditional Chinese medicine composition of this invention and the model group in terms of the four lipid levels, compared with the model group, the levels of TG, TC and LDL-C in the traditional Chinese medicine composition group of this invention show a decreasing trend, while the level of HDL-C shows an increasing trend. This indicates that the traditional Chinese medicine composition of this invention has a certain regulatory effect on the lipid abnormalities of obese rats.

[0196] One of the characteristics of obesity is a low-grade chronic inflammatory state. Therefore, this experiment selected inflammatory factors such as TNFα, CRP, IL-6, and IL-10 to detect their serum levels. By comparing the levels of inflammatory factors among different groups, the regulatory effect of the traditional Chinese medicine composition of this invention on the inflammatory state of obesity was explored. The results showed that the traditional Chinese medicine composition of this invention significantly downregulated the levels of multiple inflammatory factors such as TNFα, CRP, IL-6, and IL-10, indicating that it has a good effect on improving the inflammatory state of obese rats.

[0197] Insulin is a protein hormone secreted by pancreatic β-cells in response to endogenous or exogenous stimuli. It is the only hormone in the body that lowers blood glucose and promotes the synthesis of glycogen, fat, and protein. Insulin can also increase the uptake of glucose by peripheral tissues and promote the conversion of glucose into amino acids or fats to lower blood glucose. Insulin resistance leads to abnormally high serum insulin levels. People with insulin resistance are usually obese because a large amount of insulin is secreted but fails to exert its normal hypoglycemic effect, instead promoting fat synthesis, which further aggravates obesity and insulin resistance, creating a vicious cycle. After 8 weeks of intervention with the herbal composition of this invention, the serum insulin levels of rats in the high, medium, and low herbal groups all decreased, indicating that the insulin resistance in obese rats improved.

[0198] When excessive fat accumulates in the body, the body's mobilization and utilization rate of free fatty acids in the blood decreases, leading to the accumulation of free fatty acids. Free fatty acids are a component of blood lipids and a significant factor contributing to dyslipidemia. Under normal circumstances, the concentration of free fatty acids in the body is low. If the concentration increases, it can easily induce a series of cardiovascular diseases. Therefore, the content of free fatty acids is also one of the important indicators for assessing the degree of obesity. In this experiment, the serum free fatty acid levels in the three weight-loss groups were significantly lower than those in the model group, indicating that the herbal composition of this invention can improve dyslipidemia by regulating the content of free fatty acids.

[0199] Leptin is a hormone primarily secreted by white adipose tissue, and its serum content is directly proportional to the size of the animal's adipose tissue. Leptin acts on receptors located in the central nervous system to regulate metabolism. When an animal's body fat decreases, the serum leptin level drops significantly; conversely, when the body fat increases, the serum leptin level rises. In this experiment, the serum leptin levels in the high- and medium-dose groups were significantly lower than those in the model group, and the low-dose group also showed a certain downward trend, indicating that the herbal composition of this invention has a significant regulatory effect on serum leptin.

[0200] Obesity is closely related to adipose tissue, especially white adipose tissue. As the most abundant type of adipose tissue in the body, white adipose tissue (WAT) stores excess energy as triglycerides (TG). However, long-term excessive energy intake leads to excessive TG accumulation in adipocytes, causing rapid expansion and dysfunction of adipose tissue, ultimately resulting in obesity. Rapid expansion of WAT in the body is the most direct factor leading to obesity, and the number and volume of adipocytes are two important parameters determining the volume of white adipose tissue. Adipose tissue is the main site of energy metabolism; by utilizing lipases to hydrolyze TG to supply energy to the whole body, targeting adipose tissue is of great significance for the treatment of obesity.

[0201] This experiment used HE staining to observe the morphology of the liver and adipose tissue in different parts of rats. The results showed that the liver cells of the control group rats were normal in morphology, while the livers of the model group rats were filled with fat vacuoles of various sizes, indicating that the model group rats exhibited fatty liver. After drug intervention, the number of fat vacuoles was significantly reduced, especially in the high and medium dose groups of the traditional Chinese medicine composition of this invention, which showed particularly significant effects. This indicates that the traditional Chinese medicine composition of this invention has a strong restorative ability for fatty liver caused by a high-fat diet.

[0202] Furthermore, in the control group, the white fat cells in the groin and periepididymal region of rats were morphologically normal and uniform in size, while in the model group, compared to the control group, the cell volume was significantly larger and the size was uneven. The size of adipocytes is directly proportional to the volume of adipose tissue, and the herbal composition of this invention can significantly reduce the volume of adipocytes in obese rats, suggesting that the herbal composition of this invention promotes browning of white fat in obese rats. Compared to the control group, the brown adipose tissue of the model group rats showed many white fat cells of uneven size, indicating significant white fat accumulation in the model group rats. After intervention with the herbal composition of this invention, the brown fat cell volume was significantly reduced and the morphology became more uniform, suggesting that the herbal composition of this invention increases the activity of brown fat.

[0203] Therefore, it can be seen that the traditional Chinese medicine composition of the present invention can improve the morphology of liver cells and fat cells and reduce the accumulation of adipose tissue.

[0204] Experiment 3: Effects of the herbal composition of this invention on browning factors in the adipose tissue of obese rats

[0205] (I) Detection of mRNA expression of relevant browning factors using RT-PCR technology

[0206] 1. Materials

[0207] 1.1 Reagents, consumables and primers used in the experiment (1) Reagents and consumables

[0208]

[0209] (2) Primers: Synthesized by Beijing Tianyi Huiyuan Biotechnology Co., Ltd., information is as follows:

[0210]

[0211] 1.2 Instruments used in the experiment

[0212]

[0213] 2 methods

[0214] 2.1 RNA Extraction

[0215] RNA was extracted from the sample using Trizol, and then 100 μl of RNase-free water was added to dissolve the RNA.

[0216] 2.2 DNase I digests DNA in sample RNA

[0217]

[0218] 2.3 RNA agarose gel electrophoresis

[0219] (1) Weigh 0.45g of agarose and put it into an Erlenmeyer flask. Add 4.5ml of 10×MOPS buffer and 39.5ml of DEPC water and melt it in a microwave oven.

[0220] (2) When the temperature has cooled to about 60 degrees Celsius, add 1 ml of formaldehyde and 5 μL of GelRed, and shake well (avoid generating bubbles). Pour into a gel plate and let it solidify for 30 minutes.

[0221] (3) Take 4 μl of each RNA sample, add 2 μl of 6×RNA electrophoresis loading buffer, mix well, and add all the liquid into the denaturing gel well.

[0222] (4) Electrophoresis at 120V for 25 minutes. Observe the gel using a gel UV analyzer and photograph for preservation.

[0223] 2.4 RNA reverse transcription into cDNA

[0224] The procedure for reverse transcription of RNA into cDNA is summarized in the table below:

[0225]

[0226] 2.5 PCR detection

[0227] Take 0.2 ml thin-walled PCR tubes and label them accordingly. Add 12.5 ml of Premix Taq dye, 0.75 ml of 10 M primer mixture, and 1 ml of the corresponding cDNA to each tube. One tube without template is used as a negative control. Add water to each tube to a final volume of 25 ml. The results are summarized below:

[0228]

[0229] After mixing, the mixture was placed in a PCR instrument for pre-denaturation at 95℃ for 5 min, followed by cycles of 95℃ for 10 s, 60℃ for 30 s, and 72℃ for 30 s, for a total of 40 cycles, with a pause at 4℃. Electrophoresis was then performed at 120V for 20 min. After electrophoresis, the gel was photographed using a UV analyzer. The results are as follows: Figure 9 Electrophoresis images of PCR products. Note: M: DL2000; 1: β-actin; 2: β-actin negative control; 3: UCP-1; 4: UCP-1 negative control; 5: PGC-1α; 6: PGC-1α negative control; 7: PPARγ; 8: PPARγ negative control; 9: PRDM16; 10: PRDM16 negative control; 11: CD137; 12: CD137 negative control; 13: TMEM26; 14: TMEM26 negative control; 15: HSP70; 16: HSP70 negative control.

[0230] 2.6 Statistical Methods

[0231] Statistical analysis was performed using SPSS 26 statistical software. The data were presented as follows: The data were analyzed using the following methods: Normality was tested for each group. For comparisons between two groups where the data conformed to a normal distribution, an independent samples t-test was used. One-way ANOVA was used for comparisons among multiple groups; the LSD test and Tamhane's T2 test were used for homogeneous and unequal variances, respectively. If the data did not conform to a normal distribution, the rank-sum test for multiple independent samples was used. A p-value < 0.05 was considered statistically significant.

[0232] 3. Results

[0233] This study examined the mRNA expression levels of seven common browning markers in iWAT and eWAT, and four browning markers in BAT, respectively. Two methods were used to analyze the expression levels of these markers. -△△CT The method is to perform the calculation, and the calculation formula is as follows:

[0234] △CT (control group) = CT (target gene in control group) - CT (internal reference gene in control group)

[0235] △CT (experimental group) = CT (target gene in experimental group) - CT (internal reference gene in experimental group)

[0236] △△CT = △CT (experimental group) - △CT (control group)

[0237] 3.1 mRNA expression levels of related browning factors in iWAT

[0238] Table 3-1 Browning Factors in iWAT ΔΔCT value

[0239]

[0240] Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # indicates that compared with the blank group, # means P < 0.05, ## means P < 0.01, ### means P < 0.001; * indicates that compared with the model group, ** means P < 0.01, *** means P < 0.001.

[0241] Figure 10 Bar chart comparing the 2-ΔΔCT values ​​of various browning factors within iWAT. Note: K (blank group); M (model group); G (high-dose group); Z (medium-dose group); D (low-dose group); Y (positive group). # represents P < 0.05 compared to the blank group, ## represents P < 0.01, ### represents P < 0.001; * represents P < 0.05 compared to group M, ** represents P < 0.01, *** represents P < 0.001.

[0242] Depend on Figure 10 visible:

[0243] (1) UCP1: ① The level in the model group was significantly lower than that in the blank group, and the difference was statistically significant (P < 0.01); ② Compared with the model group, the gene expression in the high-dose group was significantly increased, and the difference was statistically significant (P < 0.001); Although there was no difference between the medium-dose group and the model group (P > 0.05), the gene expression in the medium-dose group was also upregulated; The gene expression in the low-dose group was upregulated compared with the model group, and the difference was statistically significant (P < 0.05), but the effect was not as significant as that in the high-dose group; Among the three Chinese medicine groups, the high-dose group had the most significant effect on increasing UCP1 level; ③ The gene expression in the positive group was also upregulated compared with the model group, and the difference was also statistically significant (P > 0.01), but slightly less than that in the high-dose Chinese medicine group, and there was no significant difference between the two groups (P > 0.05).

[0244] (2) PGC-1α: ① Gene expression in the model group was downregulated compared to the blank group, and there was a significant difference between the two groups (P < 0.01); ② Gene expression in all three Chinese medicine dosage groups was increased compared to the model group, but only the difference between the low-dose group and the model group was statistically significant (P > 0.05); ③ Gene expression in the positive group was significantly higher than that in the model group (P < 0.01).

[0245] (3) PPAR-γ: ① The expression of PPAR-γ in the model group was lower than that in the blank group, and there was a significant difference between the two (P < 0.05); ② All three Chinese medicine dosage groups upregulated the gene expression of PPAR-γ. Among them, there was no difference between the high-dose group and the model group (P > 0.05), while the medium and low-dose groups were significantly different from the model group (P < 0.001; P < 0.001); ③ The positive group also showed a significantly different expression from the model group (P < 0.001), and upregulated the gene expression of PPAR-γ.

[0246] (4) PRDM16: ① The gene expression in the model group was lower than that in the blank group, and there was a significant difference between the two (P<0.05); ② All three Chinese medicine dosage groups and the positive group increased the gene expression of PRDM16 in the subcutaneous white fat of obese rats. The high-dose group, the positive group and the model group were significantly different (P<0.05), while the medium and low dose groups and the model group were not different (P>0.05).

[0247] (5) CD137: ① The gene expression in the model group was significantly lower than that in the blank group, with a highly significant difference between the two (P<0.001); ② The gene expression of CD137 was increased in each treatment group compared with the model group. The differences between the high-dose group (P<0.001), medium-dose group (P<0.05), low-dose group (P<0.01), and positive group (P<0.001) and the model group were all statistically significant.

[0248] (6) TMEM26: ① Compared with the blank group, the gene expression in the model group was significantly downregulated, and the difference between the groups was extremely significant (P<0.001); ② The gene expression of TMEM26 was significantly upregulated in all treatment groups, especially the high-dose and positive control groups (P<0.001; P<0.001), followed by the low-dose group (P<0.01), and then the medium-dose group (P>0.05).

[0249] (7) HSP70: ① The gene expression in the model group was also downregulated compared with the blank group, and there was a significant difference between the two (P<0.01); ② The expression of HSP70 gene in obese rats was upregulated in all four drug administration groups. Compared with the model group, there were statistically significant differences in the high-dose group (P<0.05), low-dose group (P<0.05), and positive group (P<0.01), while there was no statistically significant difference in the medium-dose group (P>0.05).

[0250] In summary, both the herbal composition of this invention and the positive control drug orlistat upregulated the expression of browning factor within iWAT. The high-dose group of the herbal composition showed the most significant efficacy, while the positive control drug also exhibited significant efficacy, with little difference between the two groups. The medium- and low-dose groups also upregulated the gene expression of browning factor, but the effects were slightly less pronounced than those of the high-dose group and the positive control drug.

[0251] 3.2 mRNA expression levels of related browning factors within eWAT

[0252] like Figure 11 The bar chart comparing the 2-ΔΔCT values ​​of various browning factors in eWAT shows that the expression of various genes in the model group decreased significantly compared to the control group. However, after 8 weeks of drug intervention, neither the traditional Chinese medicine groups nor the positive control group had a significant regulatory effect on the expression of browning factors in visceral white adipose tissue. This suggests that the traditional Chinese medicine composition of this invention and orlistat may not have a significant regulatory effect on the visceral white adipose tissue of obese rats.

[0253] 3.3 mRNA expression levels of related browning factors within BAT

[0254] like Figure 12 Browning factors 2 within BAT -△△CT As shown in the bar chart, the expression of various genes was not significantly different among the different groups, indicating no inter-group differences. It is speculated that the herbal composition of this invention has little effect on brown adipose tissue.

[0255] (II) Detection of protein expression of relevant browning factors in iWAT using Western blotting (WB) technology

[0256] According to the RT-PCR results, only iWAT showed statistically significant differences in the expression of browning factors among the three adipose tissues. Therefore, Western blot (WB) was used to detect the corresponding protein expression of iWAT genes. Among the seven browning factors in iWAT, there was no significant difference in PGC-1α expression between the traditional Chinese medicine group and the model group. Since all commercially available TMEM26 antibodies are human antibodies, UCP1, PPAR-γ, PRDM16, CD137, and HSP70 were selected from the seven indicators for further protein expression level testing.

[0257] 1. Materials

[0258] 1.1 Antibodies

[0259]

[0260] 1.2 Reagents

[0261]

[0262] 1.3 Experimental Apparatus

[0263]

[0264] 2 methods

[0265] (1) Prepare an 8%-10% separating gel and a 5% stacking gel according to the molecular weight of the target protein.

[0266] (2) Sample loading volume of the protein to be tested: 30ul / well.

[0267] (3) Electrophoresis conditions: stacking gel at constant voltage of 90V for about 20 minutes; separating gel at constant voltage of 120V, and the electrophoresis stop time is determined by pre-stained protein markers.

[0268] (4) Wet transfer method, transfer conditions: 300mA constant current; 0.45um pore size PVDF membrane, transfer time 60min.

[0269] (5) Sealing: Immerse the membrane completely in 5% BSA-TBST and incubate on a horizontal shaker for 1 hour (RT).

[0270] (6) Primary antibody incubation: Dilute the primary antibody with 5% BSA-TBST and incubate overnight on a horizontal shaker at 4°C.

[0271]

[0272] (7) The next day, wash the membrane: wash 3 times with TBST, 10 minutes each time.

[0273] (8) Secondary antibody incubation: Dilute the secondary antibody with 5% BSA-TBST, add goat anti-rabbit IgG (H+L) HRP 1:10000, and incubate at room temperature for 1 hour. Wash the membrane: Wash the membrane 3 times with PBST, 10 minutes each time.

[0274] (9) Add ECL to the protein side of the membrane and react for 3-5 minutes; expose the film for 10-5 minutes (the exposure time is adjusted according to different light intensities), develop for 2 minutes, and fix.

[0275] (10) Analyze using IPP6 grayscale analysis software and scan with a regular scanner.

[0276] 3 Results

[0277] Western blot results showed that the protein expression of UCP1, PPAR-γ, PRDM16, HSP70, and CD137 in the model group was significantly decreased compared with the blank group; while the high-dose herbal composition of the present invention significantly upregulated the protein expression levels of UCP1, PPAR-γ, PRDM16, HSP70, and CD137; compared with the model group, the protein expression in the medium and low dose groups was also upregulated to some extent, but not as significantly as the high-dose group; the protein levels of UCP1, PPAR-γ, PRDM16, HSP70, and CD137 in the positive group were also higher than those in the model group, and the effect was better than that in the medium and low dose groups, but not as good as that in the high-dose group.

[0278] Figure 13 Bar chart showing the relative protein expression levels of various browning factors within iWAT.

[0279] Figure 14 Protein band diagram of various browning factors in iWAT.

[0280] 4 Discussion

[0281] PGC-1α is a major regulator of mitochondrial formation and oxidative metabolism, participating in the regulation of energy metabolism as a transcriptional coactivator. Both PRDM16 and PGC-1α are core transcription factors regulating brown adipose tissue differentiation and are also hallmark thermogenic factors of brown adipose tissue, inhibiting expression in white adipose tissue. PRDM16 and PGC-1α interact to jointly promote the expression of brown adipose tissue-specific genes. The stability and accumulation of PRDM16 protein in this process mainly depends on the complete activation of PPAR-γ. CD137, TMEM26, and HSP70 are all beige adipocyte-specific expression genes. All of these genes can serve as molecular markers for browning of white adipose tissue. By detecting the levels of these browning molecular markers, the degree of browning in white adipose tissue can be determined.

[0282] This experiment first used RT-PCR to detect the mRNA expression of browning molecular markers in subcutaneous white adipose tissue, visceral white adipose tissue, and beige adipose tissue. The results showed that only the subcutaneous white adipose tissue showed significant inter-group differences. In subcutaneous white adipose tissue, the herbal composition of this invention significantly upregulated the gene expression levels of UCP1, PGC-1α, PRDM16, CD137, and TMEM26, with the high-dose group showing the most significant effect. There were no differences between the visceral white adipose tissue and brown adipose tissue groups. Therefore, five factors—UCP1, PGC-1α, PRDM16, CD137, and TMEM26—were selected from inguinal white adipose tissue for further Western blotting (WB) testing to detect their protein expression. However, since there is no commercially available rat antibody for TMEM26, the detection of its protein level was abandoned. Finally, the protein expression levels of five browning factors—UCP1, PPAR-γ, PRDM16, HSP70, and CD137—were detected.

[0283] Western blot (WB) results showed that the herbal composition of this invention can effectively increase the protein expression levels of browning factors such as UCP1, PRDM16, HSP70, and CD137 in the white adipose tissue of obese rats, which is consistent with the results of gene detection. This indicates that the herbal composition of this invention can effectively promote the browning of subcutaneous white adipose tissue in obese rats, thereby achieving a weight loss effect.

[0284] Activating brown adipose tissue and inducing browning of white adipose tissue can increase energy expenditure, inhibit obesity, and improve glucose and lipid metabolism disorders and insulin resistance, which is of great significance for the prevention and treatment of obesity. Based on the results of adipocyte morphology observation by HE sectioning, PCR detection of browning factor mRNA levels, and Western blotting of inguinal white adipose tissue, the herbal composition of this invention promotes the browning of white adipose tissue and increases the activity of brown adipose tissue. This means that the herbal composition of this invention can increase the body's energy expenditure and improve obesity.

[0285] Summarize

[0286] Adipose tissue is inextricably linked to the development and progression of obesity. There are three main types of adipose tissue in the human body: white adipose tissue, brown adipose tissue, and beige adipose tissue, each differing in color, shape, distribution, and function. White adipose tissue is the most abundant type of fat in the body, and its excessive accumulation is a major characteristic of obesity. White adipose tissue stores energy and excess lipids in the form of triglycerides. Brown adipocytes are rich in mitochondria, highly express UCP1, and their primary function is to burn fat for heat. Beige adipocytes are an intermediate type of induced thermogenic adipocyte, containing a greater number of mitochondria. When stimulated by cold, exercise, or drugs, beige adipocytes within white adipocytes transform into multi-compartmental lipid droplets, their UCP1 levels increase, they express brown adipose-specific molecules, increase their respiration rate, consume fatty acids to generate heat, and exhibit characteristics of brown adipocytes.

[0287] Energy imbalance is a major cause of obesity, and reducing energy intake or increasing energy expenditure can effectively treat obesity. UCP1 can convert energy into heat, thereby increasing energy expenditure through non-shivering thermogenesis. Therefore, increasing the expression or activity of UCP1 in adipocytes may be a promising direction for the prevention and treatment of obesity. UCP1 is mainly expressed in brown and beige adipocytes, and its thermogenesis capacity can be induced in beige adipocytes in white fat cells to be equal to that of brown adipocytes. Therefore, inducing white fat to brown or activating brown fat cells to increase the expression of UCP1 in cells is a possible direction for weight loss.

[0288] With the confirmed presence of active brown adipose tissue in adults, research on brown adipose tissue has become a focus of attention for scholars, although the amount of brown adipose tissue in the adult body is very small. Later, researchers discovered beige adipocytes, and beige adipose tissue, like brown adipose tissue, has gradually become a target for inhibiting obesity. When stimulated, beige adipose tissue performs brown adipose tissue-like functions; however, when the stimulation is removed, beige adipocytes gradually lose their original characteristics, and their morphological and molecular features revert to those of white adipocytes—a bidirectional and reversible process. Therefore, targeting thermogenic fat for obesity treatment is a relatively safe and reliable direction. Activating brown adipose tissue and inducing the browning of white adipose tissue to consume more energy are key to treating obesity, improving metabolic syndrome (MS), intravascular coagulation (IR), and reducing cardiovascular risk.

[0289] Therefore, this experiment selected adipose tissue as the target. By observing morphological changes in adipose tissue and detecting the expression levels of related browning factors, it was found that the herbal composition of this invention can target adipose tissue, promoting the browning of white adipose tissue and activating the activity of brown adipose tissue in vivo by regulating the level of browning factors in white adipose tissue, thereby increasing the secretion of more UCP1 and thus increasing the body's energy consumption. This experiment verified the formulation idea of ​​the herbal composition of this invention. The experimental results prove that the herbal composition of this invention can indeed effectively reduce the accumulation of white adipose tissue and achieve weight loss in obese rats by promoting the browning of white adipose tissue and increasing energy consumption.

[0290] Orlistat is currently the only OTC weight-loss drug in the world and also the best-selling weight-loss product, with very good weight-loss effects. The experimental data show that the high-dose group of the herbal composition of this invention is comparable to, and even slightly better than, orlistat in terms of weight reduction, regulation of inflammatory states, and promotion of white adipose tissue browning. The medium and low-dose groups, however, are slightly less effective than orlistat. This demonstrates that the herbal composition of this invention (especially the high-dose group) has a good therapeutic effect on obesity.

[0291] In summary, the herbal composition of this invention can reduce body weight in obese rats, regulate the levels of blood lipids, inflammatory factors, insulin, free fatty acids, and serum leptin, and significantly improve the morphology of adipose tissue cells and increase the expression of related browning factors in inguinal white adipose tissue. These results indicate that the herbal composition of this invention can resist obesity induced by a high-fat diet. Its mechanism may be through inducing browning of white adipose tissue and increasing the activity of brown adipose tissue, with a particularly significant promoting effect on subcutaneous white adipose tissue. However, its specific mechanism of action requires further investigation.

[0292] Experiment 4: Clinical Efficacy Study of the Traditional Chinese Medicine Composition of the Invention

[0293] Experimental Overview

[0294] A total of 120 subjects were randomly assigned to either the experimental group (receiving the herbal composition of this invention) or the control group (receiving a placebo), with 60 subjects in each group (receiving at least one dose of treatment, included in the safety analysis). In the experimental group, 14 subjects dropped out (due to protocol violation, voluntary discontinuation, or other reasons), and 46 completed the 12-week follow-up. In the control group, 12 subjects dropped out (due to protocol violation, voluntary discontinuation, or other reasons), and the remaining 48 completed the 12-week follow-up.

[0295] 1 Baseline characteristics

[0296] 1.1 Age

[0297] The 120 enrolled participants were aged between 19 and 62 years. The age distribution is shown in the figure. Dividing the participants into 10-year groups, there were 41 participants aged 19-29, 39 aged 29-39, 23 aged 39-49, 13 aged 49-59, and 4 aged 59-69. Overall, the number of participants gradually decreased with increasing age. The 19-39 age group comprised 80 participants, accounting for two-thirds of the total, indicating that the recruited participants were generally young, with fewer middle-aged and elderly participants.

[0298] The mean age of the experimental group was 35.9 years (standard deviation 9.8), and the mean age of the control group was 36.7 years (11.5). The ages of the included subjects were not normally distributed. Nonparametric tests were used to compare the ages of the two groups, and the difference was not statistically significant (P = 0.586), as shown in Table 4-1.

[0299] Table 4-1 Comparison of ages between the two groups

[0300]

[0301] 1.2 Gender

[0302] Of the 120 participants, 50 were male (41.67%) and 70 were female (58.33%), indicating a predominance of female participants in this study. Both the experimental and control groups consisted of 25 males and 35 females, with no significant difference in gender distribution between the two groups.

[0303] 1.3 Geographical Location

[0304] This study recruited participants in Zhuhai, Guangdong Province; Zhengzhou, Henan Province; and Nanchang, Jiangxi Province. Participants from each center were randomly assigned to the experimental and control groups in a 1:1 ratio. 72 participants (60%) were recruited from Zhuhai, Guangdong Province; 24 participants (20%) were recruited from each of Zhengzhou, Henan Province; and 24 participants (20%) were recruited from each of Nanchang, Jiangxi Province.

[0305] 1.1.1 Weight and BMI

[0306] The subjects' average weight was 87.0 kg (18.4), and their BMI was 30.9 kg / m². 2 (4.7); The average weight of the control group was 87.5 kg (18.4), and the BMI was 31.5 kg / m². 2 (4.5). Table 4-2 compares the weight and BMI between the two groups. Since the data do not conform to a normal distribution, a nonparametric test was used. The results showed no significant differences in weight and BMI between the two groups (P > 0.05). Furthermore, stratified comparisons by gender showed no statistically significant differences (P > 0.05).

[0307] Table 4-2 Comparison of baseline weight and BMI between the two groups

[0308]

[0309] A comparison of the weight and BMI of all male and female participants revealed that the average weight of men was 103.0 kg (17.6), and their average BMI was 33.8 kg / m². 2 (5.43); the average weight of women was 77.1 kg (9.4), and the BMI was 29.5 kg / m². 2 (2.9). Men had significantly higher body weight and BMI than women (P < 0.001), as shown in Table 4-3.

[0310] Table 4-3 Comparison of male and female weight and BMI

[0311]

[0312] 1.1.2 Body fat percentage and lean body mass percentage

[0313] The average body fat percentage in the experimental group was 34.1% (5.3), with an average body fat percentage of 30.0% (5.3) for men and 36.7% (3.4) for women, and an average lean body mass of 65.9% (5.3). The average body fat percentage in the control group was 34.3% (5.4), with an average body fat percentage of 29.2% (4.2) for men and 37.6% (2.9) for women, and an average lean body mass of 65.7% (5.4). Body fat percentage and lean body mass followed a normal distribution. A t-test was used to compare the overall body fat percentage, lean body mass, and body fat percentage of men and women between the two groups. No statistically significant differences were found (P > 0.05), as shown in Table 4-4.

[0314] Table 4-4 Comparison of body fat percentage and lean body mass percentage between the two groups

[0315]

[0316] The body fat percentages of all male and female subjects were compared. The average body fat percentage of males was 29.6% (4.7%), and the average body fat percentage of females was 37.1% (3.2%). The difference between the two groups was significant (P < 0.001), indicating that the body fat percentage of females was significantly higher than that of males (see Table 4-5).

[0317] Table 4-5 Comparison of Body Fat Percentage between Men and Women

[0318]

[0319] 1.1.3 Waist circumference, hip circumference, and waist-to-hip ratio (WHR)

[0320] Waist circumference, hip circumference, and waist-to-hip ratio were compared between the two baseline groups. The mean waist circumference in the experimental group was 100.7 cm (12.6), hip circumference was 109.8 cm (8.9), and waist-to-hip ratio was 0.92 (0.07); the mean waist circumference in the control group was 101.8 cm (13.3), hip circumference was 110.5 cm (8.5), and waist-to-hip ratio was 0.92 (0.07). The data did not conform to a normal distribution, and nonparametric tests were used. There were no statistically significant differences in waist circumference, hip circumference, and waist-to-hip ratio between the two groups (P > 0.05), as shown in Tables 4-6.

[0321] Table 4-6 Comparison of waist and hip circumference between the two groups

[0322]

[0323] Waist circumference, hip circumference, and waist-to-hip ratio were compared between all male and female subjects. The average waist circumference of males was 112 cm (11.0), hip circumference was 114.9 cm (9.5), and waist-to-hip ratio was 0.97 (0.04); the average waist circumference of females was 94.2 cm (8.5), hip circumference was 107.1 cm (6.4), and waist-to-hip ratio was 0.88 (0.06). There were significant differences between the two groups (P < 0.001), indicating that the waist circumference, hip circumference, and waist-to-hip ratio of males were significantly larger than those of females (see Table 4-7).

[0324] Table 4-7 Comparison of waist circumference, hip circumference, and waist-to-hip ratio between men and

[0325]

[0326] 1.4 Blood Pressure

[0327] The systolic and diastolic blood pressures of the two groups were compared. They conformed to a normal distribution and had homogeneous variances. The T-test showed that there was no significant difference between the two groups (P>0.05), as shown in Table 4-8.

[0328] Table 4-8 Comparison of blood pressure between the two groups

[0329]

[0330] 1.1.4 Other test indicators

[0331] The laboratory test indicators of the two groups, including FPG, HbA1c, CP, FINS, hsCRP, UA, CR, BUN, ALT, and AST, were compared. None of them conformed to a normal distribution, so nonparametric tests were used. The results showed that there were no statistically significant differences between the two groups for the above indicators (P > 0.05), as shown in Table 4-9.

[0332] Comparison of laboratory test indicators in groups 4-9

[0333]

[0334] 1.1.5 Complications

[0335] By statistically analyzing the number of outliers in each indicator of the baseline data and using the chi-square test, the results showed that there was no statistically significant difference in the number of patients with complications between the two groups (P>0.05), as shown in Table 4-10.

[0336] Table 4-10 Comparison of the number of people with hypertension in the two groups

[0337]

[0338] 1.5 Syndrome Distribution

[0339] According to the diagnostic criteria of Traditional Chinese Medicine Internal Medicine, obesity has five syndrome types: stomach heat and fire stagnation, phlegm-dampness accumulation, qi stagnation and blood stasis, spleen deficiency and indigestion, and spleen and kidney yang deficiency. Statistical analysis showed that the syndrome types among the subjects included in this study, from most to least prevalent, were: phlegm-dampness accumulation (59 subjects, 49.2%), spleen deficiency and indigestion (21 subjects, 17.5%), stomach heat and fire stagnation (15 subjects, 12.5%), spleen and kidney yang deficiency (15 subjects, 12.5%), and qi stagnation and blood stasis (10 subjects, 8.3%).

[0340] Table 4-11 Comparison of the distribution of TCM syndromes in the two groups

[0341]

[0342] Note: Because percentages are calculated by rounding, the sum may not equal 100%.

[0343] 2. Statistical analysis of outcome indicators

[0344] 2.1 Primary therapeutic indicators

[0345] The primary efficacy endpoint of this study was the weight change from baseline in obese subjects after 12 weeks of intervention with the herbal composition and placebo of this invention. A two-way repeated measures covariance analysis was used, with baseline weight as a covariate. Based on Cook distance results and clinical epidemiological knowledge, no outliers requiring special handling were found; the residuals followed a normal distribution with homogeneous variances; the sphericity test indicated that the football-shaped hypothesis was not met (W = 0.860, P = 0.001), and the results corrected using the Hindfield method were accepted; an interaction between group and time was observed (F...). 时间*组别 =52.782, P<0.001), therefore, a single-effects analysis was performed, as shown in Table 4-12.

[0346] Table 4-12 Comparison of weight covariance analysis between the two groups

[0347]

[0348] The time-dependent effects analysis was performed pairwise using the Bonfroni method. The results showed that, with the intervention of the herbal composition of this invention, weight gradually decreased at weeks 8 and 12 compared to week 4, with statistically significant differences (P<0.001); weight also decreased at week 12 compared to week 8, with statistically significant differences (P<0.001). With the placebo intervention, there were no statistically significant differences in weight change at weeks 8 and 12 compared to week 4 (P<0.001).

[0349] The one-way effect analysis of the groups showed no significant main effect (P = 0.194). At week 4, the difference in weight change between the two groups was 1.70 kg (95% confidence interval -5.575, 8.981), and at week 8, the difference was 4.59 kg (-2.374, 11.959). There were no statistically significant differences in weight change between the two groups at weeks 4 and 8 (P = 0.643, P = 0.194). At week 12, the difference in weight change between the two groups further increased to 7.55 kg (0.672, 14.429), which was statistically significant (P = 0.032). Figure 15 The changes in body weight were shown in the experimental and control groups.

[0350] The experimental group experienced an average weight loss of 6.23% (4.92, 7.55), while the control group experienced a weight gain of 1.50% (0.72, 2.27). 67.4% of the participants in the experimental group (31 people) experienced a weight loss of more than 5%, compared to 1 person (1.7%) in the control group. Seven people (7.4%) in the experimental group experienced a weight loss of more than 10%.

[0351] Table 4-13 Percentage of weight loss in the two groups

[0352]

[0353] Table 4-14 compares the weight changes of men and women in the experimental group. The overall weight of both men and women in the experimental group showed a normal distribution, and a t-test was used for analysis. The results showed that in the experimental group, men weighed 102.9 kg (16.5 kg) at week 0 and 93.9 kg (13.2 kg) at week 12, a weight loss of 9.07 kg (5.23 kg). Compared with week 0, men's weight decreased significantly at week 12 (P < 0.001). Women weighed 76.8 kg (10.9 kg) at week 0 and 73.1 kg (10.2 kg) at week 12, a weight loss of 3.74 kg (3.34 kg). Compared with week 0, women's weight decreased significantly at week 12 (P < 0.001). Compared with women, men's weight was significantly greater than women's (P < 0.001), and the weight loss effect of men was significantly better than that of women (P < 0.001).

[0354] Table 4-14 Comparison of body weight between males and females in the experimental group (unit: kg)

[0355]

[0356] Note: Difference 12 / 0 P 12 / 0 The table below shows the differences and p-values ​​between the 12-week and 0-week groups.

[0357] 2.2 BMI

[0358] Table 4-15 compares the changes in BMI from 0 to 12 weeks between the two groups. The BMIs of the two groups are not normally distributed, and a nonparametric test was used. The BMI of the experimental group at week 0 was 30.9 kg / m². 2 (29.5, 32.3), 12-week BMI is 28.9 kg / m². 2 (27.8, 30.0), BMI decreased by 2.0 kg / m². 2 (1.5, 2.5), the difference was statistically significant (P < 0.001); the BMI of the control group at week 0 was 31.5 kg / m². 2 (30.2, 32.8), 12-week BMI is 31.9 kg / m². 2 (30.7, 33.2), BMI increased by 0.4 kg / m². 2 (0.2, 0.7), the difference was statistically significant (P < 0.001). There was no statistically significant difference in BMI between the two groups at weeks 0 and 4 (P > 0.05), but at weeks 8 (P = 0.028) and 12 (P < 0.001), the experimental group had a significantly smaller BMI than the control group, and at week 12, the experimental group's BMI decreased significantly more than that of the control group (P < 0.001).

[0359] Table 4-15 BMI Changes in Two Groups (Unit: kg / m²) 2 )

[0360]

[0361] Table 4-16 compares the BMI of males and females in the experimental group. The BMI follows a normal distribution, and a t-test was used. The BMI of males in the experimental group at week 0 was 33.6 kg / m². 2 (5.3), at 12 weeks, it was 30.6 kg / m 2 (4.1) BMI decreased by 2.97 kg / m². 2 (1.76), the difference was statistically significant (P<0.001); the BMI of women at week 0 was 29.2 kg / m². 2 (3.2), at 12 weeks, it was 27.8 kg / m 2 (3.2) BMI decreased by 1.41 kg / m². 2(1.25), the difference was statistically significant (P < 0.001). Compared with women, men had a significantly larger BMI (P < 0.05), and men experienced a significantly greater decrease in BMI than women (P = 0.001).

[0362] Table 4-16 Comparison of BMI between males and females in the experimental group (unit: kg / m²) 2 )

[0363]

[0364] 2.3BFP

[0365] Table 4-17 compares the changes in body fat percentage between the two groups. The body fat percentages followed a normal distribution, and a t-test was used for analysis. In the experimental group, the body fat percentage was 34.1 (5.3%) at week 0 and 31.9 (5.6%) at week 12, a decrease of 2.2 (2.4%), which was statistically significant (P < 0.001). In the control group, the body fat percentage was 34.3 (5.4%) at week 0 and 35.2 (5.4%) at week 12, an increase of 0.96 (2.0%), which was statistically significant (P = 0.002). There were no statistically significant differences in body fat percentage between the two groups at weeks 0, 4, and 8 (P > 0.05). At week 12, the BFP in the experimental group was significantly lower than that in the control group (P < 0.001), and the decrease in BFP in the experimental group was significantly greater than that in the control group (P = 0.001).

[0366] Table 4-17 Changes in body fat percentage between the two groups (unit: %)

[0367]

[0368]

[0369] Table 4-18 shows the stratified comparison of body fat percentage (BFP) in the experimental groups by gender. Both male and female BFP followed a normal distribution, and a t-test was used for analysis. In male subjects, the BFP at week 0 was 30.0% (5.3), and at week 12 it was 27.4% (4.2), a decrease of 2.59% (2.27), which was statistically significant (P < 0.001). In female subjects, the BFP at week 0 was 36.7% (3.4), and at week 12 it was 34.8% (4.3), a decrease of 1.87% (2.46), which was also statistically significant (P < 0.001). Compared with males, females had a significantly higher body fat percentage (P < 0.001), but there was no significant difference in the decrease in body fat percentage between the two groups (P = 0.319).

[0370] Table 4-18 Comparison of body fat percentage between males and females in the experimental group (unit: %)

[0371]

[0372] 2.4LBP

[0373] Table 4-19 compares the changes in LBP between the two groups. LBP follows a normal distribution, and a t-test was used for analysis. In the experimental group, LBP at week 0 was 65.9% (5.3), and at week 12 it was 68.1% (5.6), an increase of 2.2% (2.4), which was statistically significant (P < 0.001). In the control group, LBP at week 0 was 65.7% (5.4), and at week 12 it was 64.8% (5.4), a decrease of 1.0% (2.0), which was statistically significant (P = 0.002). There were no statistically significant differences in LBP between the two groups at weeks 0, 4, and 8 (P > 0.05), but at week 12, the LBP in the experimental group was significantly higher than that in the control group (P < 0.001). Because LBP = 100% - BFP, and the change in LBP is opposite to that of BFP, LBP was not compared by gender.

[0374] Table 4-19 Comparison of LBP between the two groups (unit: %)

[0375]

[0376] 2.5WC

[0377] Table 4-20 compares the changes in WC between the two groups. WC followed a normal distribution with homogeneous variance, and a t-test was used for analysis. In the experimental group, WC was 100.7 cm (12.6) at week 0 and 96.3 cm (10.7) at week 12, a decrease of 4.4 cm (3.6), which was statistically significant (P < 0.001). In the control group, WC was 101.8 cm (13.3) at week 0 and 102.8 cm (12.5) at week 12, an increase of 1.0 cm (5.1), but the difference was not statistically significant (P = 0.165). There were no statistically significant differences in WC between the two groups at weeks 0, 4, and 8 (P > 0.05). At week 12, the experimental group's WC was significantly smaller than the control group's (P < 0.001), and the decrease in WC in the experimental group was significantly greater than that in the control group (P < 0.001).

[0378] Table 4-20 Comparison of WC between the two groups (unit: cm)

[0379]

[0380] Table 4-21 compares the WC values ​​of males and females in the experimental group. The values ​​conform to a normal distribution, and a t-test was used. In the experimental group, the WC of males at week 0 was 111.6 cm (10.4), and at week 12 it was 104.8 cm (8.7), a decrease of 6.82 cm (2.87), which was statistically significant (P < 0.001). In females, the WC at week 0 was 93.7 cm (8.2), and at week 12 it was 90.9 cm (8.0), a decrease of 2.77 cm (3.06), which was also statistically significant (P < 0.001). Compared with females, males had significantly larger WC values ​​(P < 0.05), and the decrease in WC was significantly greater in males than in females (P < 0.001).

[0381] Table 4-21 Comparison of WC between males and females in the experimental group (unit: cm)

[0382]

[0383] 2.6HC

[0384] Table 4-22 compares the changes in HC between the two groups. HC followed a normal distribution with homogeneous variances, and a t-test was used for analysis. In the experimental group, HC was 109.8 cm (8.9) at week 0 and 106.9 cm (7.3) at week 12, a decrease of 2.9 cm (3.1), which was statistically significant (P < 0.001). In the control group, HC was 110.5 cm (8.5) at week 0 and 111.9 cm (8.3) at week 12, an increase of 1.4 cm (3.0), which was statistically significant (P = 0.003). There was no statistically significant difference in HC between the two groups at weeks 0, 4, and 8 (P > 0.05). At week 12, the HC in the experimental group was significantly lower than that in the control group (P < 0.001), and the decrease in HC in the experimental group was significantly greater than that in the control group (P < 0.001).

[0385] Table 4-22 Comparison of HC between the two groups (unit: cm)

[0386]

[0387]

[0388] Table 4-23 compares the HC values ​​of males and females in the experimental group. The values ​​conform to a normal distribution, and a t-test was used. In the experimental group, the HC value of males at week 0 was 114.6 cm (8.9) and at week 12 it was 110.1 cm (6.3), a decrease of 4.47 cm (3.74), which was statistically significant (P < 0.001). In females, the HC value at week 0 was 106.7 cm (7.6) and at week 12 it was 104.9 cm (7.3), a decrease of 1.83 cm (2.06), which was also statistically significant (P < 0.001). Compared with females, males had significantly higher HC values ​​(P < 0.05), and the decrease in HC value was significantly greater in males than in females (P = 0.004).

[0389] Table 4-23 Comparison of HC between males and females in the experimental group (unit: cm)

[0390]

[0391] 2.7 WHR

[0392] Table 4-24 compares the changes in WHR between the two groups. Body fat percentages followed a normal distribution with homogeneous variances, and a t-test was used for analysis. In the experimental group, WHR was 0.916 (0.068) at week 0 and 0.900 (0.065) at week 12, a decrease of 0.016 (0.027), which was statistically significant (P < 0.001). In the control group, WHR was 0.919 (0.074) at week 0 and 0.917 (0.065) at week 12, a decrease of 0.002 (0.034), but the difference was not statistically significant (P = 0.716). There were no statistically significant differences in WHR between the two groups at weeks 0, 4, 8, and 12 (P > 0.05), but the decrease in WHR in the experimental group was significantly greater than that in the control group (P = 0.032).

[0393] Table 4-24 Comparison of WHR between the two groups

[0394]

[0395] Table 4-25 compares the whr h of males and females in the experimental group. The results conform to a normal distribution, and a t-test was used. In the experimental group, the whr h of males at week 0 was 0.973 (0.029) and at week 12 was 0.950 (0.035), a decrease of 0.023 (0.028), which was statistically significant (P = 0.003). In females, the whr h of females at week 0 was 0.879 (0.059) and at week 12 was 0.867 (0.059), a decrease of 0.011 (0.026), which was also statistically significant (P = 0.032). Compared with females, males had significantly higher whr h (P < 0.05), while there was no significant difference in the decrease in whr h between males and females (P = 0.166).

[0396] Table 4-25 Comparison of WHR between males and females in the experimental group

[0397]

[0398] 2.8FPG

[0399] Table 4-26 compares the changes in FPG between the two groups. Since FPG did not conform to normality, non-parametric tests were used for analysis. In the experimental group, FPG was 5.75 mmol / L (5.21, 6.29) at week 0 and 5.31 mmol / L (5.10, 5.52) at week 12, a decrease of 0.44 mmol / L (-0.41, 0.92), which was not statistically significant (P = 0.67). In the control group, FPG was 5.40 mmol / L (5.18, 5.63) at week 0 and 5.86 mmol / L (5.44, 6.27) at week 12, an increase of 0.46 mmol / L (0.04, 0.87), which was statistically significant (P = 0.001). There was no statistically significant difference in FPG between the two groups at week 0 (P = 0.420). At week 12, the FPG in the experimental group was significantly lower than that in the control group (P = 0.005), and the decrease in FPG in the experimental group was significantly greater than that in the control group (P = 0.025).

[0400] Table 4-26 Comparison of FPG between the two groups (unit: mmol / L)

[0401]

[0402] 2.9HbA1c

[0403] Table 4-27 compares the changes in HbA1c between the two groups. Since HbA1c does not conform to normality, nonparametric tests were used for analysis. In the experimental group, HbA1c was 5.75% (5.41, 6.09) at week 0 and 5.40% (5.23, 5.57) at week 12, a decrease of 0.35% (0.11, 0.60), which was statistically significant (P = 0.015). In the control group, HbA1c was 5.54% (5.37, 5.71) at week 0 and 5.77% (5.50, 6.04) at week 12, an increase of 0.24% (0.01, 0.46), which was also statistically significant (P = 0.002). There was no statistically significant difference in HbA1c between the two groups at week 0 (P = 0.979). At week 12, the HbA1c in the experimental group was significantly lower than that in the control group (P = 0.007), and the decrease in HbA1c in the experimental group was significantly greater than that in the control group (P < 0.001).

[0404] Table 4-27 Comparison of HBA1c between the two groups (unit: %)

[0405]

[0406] 2.10CP

[0407] Table 4-28 compares the changes in CP between the two groups of subjects. Since CP does not conform to normality, nonparametric tests were used for analysis. In the experimental group, CP was 1.17 nmol / L (1.01, 1.33) at week 0 and 1.00 nmol / L (0.87, 1.12) at week 12, a decrease of 0.17 nmol / L (0.07, 0.27), which was statistically significant (P = 0.001). In the control group, CP was 1.15 nmol / L (1.00, 1.30) at week 0 and 1.14 nmol / L (1.01, 1.27) at week 12, a decrease of 0.01 nmol / L (-0.10, 0.12), which was not statistically significant (P = 0.731). There were no statistically significant differences in CP between the two groups at week 0 (P = 0.91) and week 12 (P = 0.081), and the decrease in CP between the two groups was not significantly different (P = 0.072).

[0408] Table 4-28 Comparison of CP between the two groups (unit: nmol / L)

[0409]

[0410] 2.11FINS

[0411] Table 4-29 compares the changes in FINS between the two groups of subjects. Since FINS does not conform to normality, non-parametric tests were used for analysis. In the experimental group, FINS was 152.0 pmol / L (120.0, 184.0) at week 0 and 127.2 pmol / L (99.0, 155.4) at week 12, a decrease of 24.8 pmol / L (6.33, 43.3), which was statistically significant (P = 0.010). In the control group, FINS was 145.6 pmol / L (114.4, 176.7) at week 0 and 134.9 pmol / L (106.1, 163.7) at week 12, a decrease of 10.7 pmol / L (-12.5, 33.8), but the difference was not statistically significant (P = 0.389). There was no statistically significant difference in FINS between the two groups at week 0 (P = 0.919) and week 12 (P = 0.602) (P > 0.05), and there was no statistically significant difference in the decrease in FINS between the two groups (P = 0.586).

[0412] Table 4-29 Comparison of FINS between the two groups (unit: pmol / L)

[0413]

[0414] 2.12hsCRP

[0415] Table 4-30 compares the changes in hsCRP between the two groups. Since hsCRP does not conform to normality, nonparametric tests were used for analysis. In the experimental group, hsCRP was 3.17 mg / L (2.28, 4.06) at week 0 and 2.29 mg / L (1.33, 3.25) at week 12, a decrease of 0.88 mg / L (-0.40, 2.15), which was statistically significant (P = 0.004). In the control group, hsCRP was 3.05 mg / L (1.91, 4.20) at week 0 and 2.99 mg / L (1.79, 4.20) at week 12, a decrease of 0.06 mg / L (-0.98, 1.09), but the difference was not statistically significant (P = 0.329). There was no statistically significant difference in hsCRP levels between the two groups at week 0 (0.201) and week 12 (0.899), and there was no statistically significant difference in the decrease in hsCRP levels between the two groups (P = 0.246).

[0416] Table 4-30 Comparison of hsCRP levels between the two groups (unit: mg / L)

[0417]

[0418] 2.13UA

[0419] Table 4-31 compares the changes in UA in the two groups of subjects. UA does not conform to normality, and non-parametric tests were used for analysis. In the experimental group, UA was 391.0 μmol / L (360.9, 421.0) at week 0 and 345.7 μmol / L (319.3, 372.2) at week 12, a decrease of 45.2 μmol / L (20.5, 69.9), which was statistically significant (P<0.001). In the control group, UA was 396.6 μmol / L (364.1, 429.2) at week 0 and 407.1 μmol / L (374.3, 439.9) at week 12, an increase of 10.5 μmol / L (7.8, 28.8), but the difference was not statistically significant (P=0.359). There was no statistically significant difference in UA between the two groups at week 0 (P = 0.806). At week 12, the UA in the experimental group was significantly lower than that in the control group (P = 0.01), and the decrease in UA in the experimental group was significantly greater than that in the control group (P < 0.001).

[0420] Table 4-31 Comparison of UA between the two groups (unit: μmol / L)

[0421]

[0422] 2.14CR

[0423] Table 4-32 compares the changes in CR (response rate) between the two groups. Since the CR did not conform to normality, non-parametric tests were used for analysis. In the experimental group, the CR at week 0 was 66.2 μmol / L (61.6, 70.9) and at week 12 it was 62.0 μmol / L (57.3, 66.8), a decrease of 4.22 μmol / L (-0.09, 8.53), which was statistically significant (P = 0.003). In the control group, the CR at week 0 was 65.1 μmol / L (60.8, 69.4) and at week 12 it was 62.7 μmol / L (57.9, 67.5), a decrease of 2.4 μmol / L (-1.62, 6.42), but the difference was not statistically significant (P = 0.412). There was no statistically significant difference in CR between the two groups at week 0 (P = 0.625) and week 12 (P = 0.482), and there was no statistically significant difference in the decrease in CR between the two groups (P = 0.097).

[0424] Table 4-32 Comparison of CR between the two groups (unit: μmol / L)

[0425]

[0426] 2.15BUN

[0427] Table 4-33 compares the changes in BUN in the two groups of subjects. Since BUN did not conform to normality, non-parametric tests were used for analysis. In the experimental group, BUN was 4.58 mmol / L (4.16, 5.00) at week 0 and 4.21 mmol / L (3.91, 4.52) at week 12, a decrease of 0.36 mmol / L (-0.08, 0.80), with no statistically significant difference (P = 0.075). In the control group, BUN was 4.32 mmol / L (4.03, 4.61) at week 0 and 5.33 mmol / L (3.18, 7.48) at week 12, an increase of 1.01 mmol / L (1.17, 3.19), with no statistically significant difference (P = 0.983). There were no statistically significant differences in BUN levels between the two groups at week 0 (P = 0.596) and week 12 (P = 0.390), and there was no statistically significant difference in the decrease in BUN between the two groups (P = 0.197).

[0428] Table 4-33 Comparison of BUN between the two groups (unit: mmol / L)

[0429]

[0430] 2.16ALT

[0431] Table 4-34 compares the changes in ALT between the two groups of subjects. Since ALT levels did not conform to normality, nonparametric tests were used for analysis. In the experimental group, ALT was 45.5 U / L (32.2, 58.8) at week 0 and 28.7 U / L (21.3, 36.0) at week 12, a decrease of 16.8 U / L (6.9, 26.7), which was statistically significant (P < 0.001). In the control group, ALT was 41.6 U / L (30.8, 52.3) at week 0 and 41.9 U / L (30.8, 53.0) at week 12, an increase of 0.3 U / L (-7.0, 7.6), but the difference was not statistically significant (P = 0.704). There was no statistically significant difference in ALT between the two groups at 0 (P = 0.952). At 12 weeks, ALT in the experimental group was significantly lower than that in the control group (P = 0.009), and the decrease in ALT in the experimental group was significantly greater than that in the control group (P = 0.005).

[0432] Table 4-34 Comparison of ALT levels between the two groups (unit: U / L)

[0433]

[0434] 2.17AST

[0435] Table 4-35 compares the changes in AST in the two groups of subjects. Since AST does not conform to normality, nonparametric tests were used for analysis. In the experimental group, AST was 30.3 U / L (21.3, 39.4) at week 0 and 23.6 U / L (18.3, 28.9) at week 12, a decrease of 6.7 U / L (-1.1, 14.5), which was statistically significant (P = 0.002). In the control group, AST was 27.6 U / L (22.5, 32.8) at week 0 and 28.6 U / L (22.4, 34.8) at week 12, an increase of 1.0 U / L (-3.7, 5.7), but the difference was not statistically significant (P = 0.550). There was no statistically significant difference in AST between the two groups at week 0 (P = 0.702), but at week 12, the AST in the experimental group was significantly smaller than that in the control group (P = 0.028), and there was no statistically significant difference in the change of AST between the two groups (P = 0.094).

[0436] Table 4-35 Comparison of AST between the two groups (unit: U / L)

[0437]

[0438] 2.18SBP

[0439] Table 4-36 compares the changes in SBP between the two groups of subjects. SBP followed a normal distribution, and a t-test was used for analysis. In the experimental group, SBP was 128.0 mmHg (18.9) at week 0 and 124.4 mmHg (120.0) at week 12, a decrease of 3.5 mmHg (12.9), with no statistically significant difference (P = 0.070). In the control group, SBP was 126.1 mmHg (16.0) at week 0 and 124.5 mmHg (14.6) at week 12, a decrease of 1.6 mmHg (11.0), with no statistically significant difference (P = 0.323). There was no statistically significant difference in SBP between the two groups at week 0 (P = 0.609) and week 12 (P = 0.972), and no statistically significant difference in the decrease in SBP between the two groups (P = 0.430).

[0440] Table 4-36 Comparison of SBP between the two groups (unit: mmHg)

[0441]

[0442] 2.19DBP normal distribution

[0443] Table 4-37 compares the changes in DBP between the two groups of subjects. DBP followed a normal distribution, and a t-test was used for analysis. In the experimental group, DBP was 84.0 mmHg (12.1) at week 0 and 83.1 mmHg (12.2) at week 12, a decrease of 0.9 mmHg (10.6), with no statistically significant difference (P = 0.582). In the control group, DBP was 85.1 mmHg (11.7) at week 0 and 84.6 mmHg (12.9) at week 12, a decrease of 0.5 mmHg (7.2), with no statistically significant difference (P = 0.620). There was no statistically significant difference in DBP between the two groups at week 0 (P = 0.641) and week 12 (P = 0.565) (P > 0.05), and no statistically significant difference in the decrease in DBP between the two groups (P = 0.852).

[0444] Table 4-37 Comparison of DBP between the two groups (unit: mmHg)

[0445]

[0446] 3 Results

[0447] The aforementioned clinical trial indicators included: weight, body mass index (BMI), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body fat percentage (BFP), lean body mass percentage (LBP), fasting plasma glucose (FPG), glycated hemoglobin (HbA1c), fasting C-peptide (CP), fasting insulin (FINS), high-sensitivity C-reactive protein (hsCRP), uric acid (UA), creatinine (CR), blood urea nitrogen (BUN), liver serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), systolic blood pressure (SBP), and diastolic blood pressure (DBP). Patients experienced significant effects after taking the herbal composition of this invention.

[0448] Clinical applications have revealed that, in addition to weight loss, the herbal composition of this invention also improves sleep, regulates glucose and lipid metabolism, lowers uric acid, regulates blood pressure, and improves emotional disorders.

[0449] 4 Typical Cases

[0450] Patient Han, male, 19 years old.

[0451] Chief complaint at initial consultation: obesity, insomnia, and difficulty falling asleep.

[0452] Medical history: Height 192cm, weight 163kg, rapid weight gain over the past three years, approximately 15kg per year, BMI 47.63.

[0453] The patient experiences difficulty falling asleep, taking approximately 120 minutes, and only sleeps for 2 hours. They report a one-year history of rectal bleeding due to internal hemorrhoids, and generally have difficulty thinking clearly and communicating with others. Their appetite is fair, but their urine is cloudy and their stools are sticky. Their tongue is dark red with a slightly yellow coating, and their pulse is slippery and slightly rapid.

[0454] Urinalysis over the past six months showed: occult blood in urine (+), protein in urine (+-), and 19.6 red blood cells in urine. Physical examination at the Affiliated Hospital of Qingdao University showed: uric acid 581 (89.2-416) umol / L, insulin 43.24 (2.6-24.9) uIU / ml. Fatty liver for 4 years, high blood pressure for 6 years, BP 190 / 110 mmHg, currently taking perindopril 8mg qd and metoprolol extended-release tablets 47.5mg qd. Father and grandfather have a history of hypertension. The patient reports a highest blood pressure of 180 / 105 mmHg.

[0455] The traditional Chinese medicine composition of the present invention was administered in a total of 30 doses.

[0456] At the second visit, weight had decreased to 137 kg, blood pressure was 160 / 90 mmHg, and uric acid was 470-480 umol / L. At the third visit, weight had decreased to 125 kg. At the fourth visit, weight was 110 kg, and hypertension had significantly improved.

[0457] At the sixth visit, the patient's weight had decreased to 102 kg. Western antihypertensive medication was discontinued, and the patient was only taking traditional Chinese medicine. Blood pressure was controlled at 140-150 / 80-90 mmHg. Tests at a local hospital showed normal blood uric acid and fasting insulin levels. Self-measured blood pressure was 150 / 90 mmHg. A follow-up phone call revealed that after taking the medication, the patient's blood pressure was 150 / 80 mmHg, falling asleep faster, and fatigue symptoms had lessened.

Claims

1. A traditional Chinese medicine composition for weight loss, characterized in that, It is made from the following Chinese medicinal materials in parts by weight: raw astragalus 30-45 parts, prepared atractylodes 10-15 parts, cinnamon 15-23 parts, dried tangerine peel 20-30 parts, lotus leaf 30-45 parts, raw hawthorn 20-30 parts, kelp 15-23 parts, turmeric 10-15 parts, stir-fried radish seed 10-15 parts, and raw cattail pollen 10-15 parts.

2. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 1, characterized in that, Includes the following steps: The above-mentioned weight proportions of Chinese medicinal materials were extracted by decoction with water.

3. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 2, characterized in that, The process includes the following steps: soaking the above-mentioned weight proportions of Chinese medicinal materials in water, decocting them, and concentrating them to obtain the final product.

4. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 2, characterized in that, The water decoction extraction is performed twice.

5. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 2, characterized in that, The process includes the following steps: Soak the above-mentioned weight proportions of Chinese medicinal herbs in 10-15 times their weight of water for 0.5-1 hour, then bring to a boil over high heat. After boiling, reduce to a simmer and continue simmering for 30-40 minutes. Pour out the first decoction. For the second decoction, add 8-12 times their weight of water and continue simmering. After boiling, simmer for 20-30 minutes. Pour out the second decoction. Combine the two decoctions. Heat and concentrate the combined decoction, then refrigerate it.

6. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 5, characterized in that, The process includes the following steps: Soak the above-mentioned weight of Chinese medicinal herbs in 10 times their weight of water for 1 hour, then bring to a boil over high heat. After boiling, reduce to a simmer and continue simmering for 30 minutes. Pour out the first decoction. For the second decoction, add 8 times the weight of water of the Chinese medicinal herbs and continue simmering. After boiling, simmer for 30 minutes. Pour out the second decoction. Combine the two decoctions. Heat and concentrate the combined decoction, then refrigerate it.

7. The method for preparing a weight-loss traditional Chinese medicine composition according to claim 5, characterized in that, The heating and concentration process is carried out at a temperature of 80-100℃, to a concentration of 0.8-3.5 g / ml. -1 .

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