Traditional Chinese medicine preparation based on black tea and application of traditional Chinese medicine preparation in improvement of simple obesity

Abstract

The invention discloses a traditional Chinese medicine preparation based on black tea and application of the traditional Chinese medicine preparation to improvement of simple obesity. The traditional Chinese medicine preparation is prepared from the following components in parts by weight: 10-20 parts of black tea, 5-10 parts of raspberry, 5-10 parts of fructus alpiniae oxyphyllae and 5-10 parts of hawthorn. The traditional Chinese medicine preparation based on black tea disclosed by the invention not only can well relieve glucose and lipid metabolism hormone disorder, but also can effectively relieve body oxidative stress injury and obesity-related low-degree inflammation states, and has a good potential application value.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine preparation based on black tea and its use in improving simple obesity, belonging to the technical field of traditional Chinese medicine. Background Art

[0002] Glucolipid metabolic hormone disorder is a common metabolic imbalance state induced by obesity, mainly manifested as abnormal secretion or action of various metabolic regulatory hormones such as insulin, leptin, adiponectin, etc. This hormone disorder can lead to the imbalance of blood glucose and blood lipid regulation mechanisms, mainly manifested as abnormal glucose tolerance, insulin resistance, leptin resistance, etc. At the same time, obesity will also induce the occurrence of oxidative stress in the body. Under a high-fat diet or metabolic disorder state, the level of free fatty acids in the body increases, mitochondrial function is damaged, and the production of reactive oxygen species (ROS) increases. The activities of SOD and GSH-Px in the body are inhibited due to excessive ROS load, thus weakening the ability to scavenge free radicals and leading the body to be in a state of oxidative stress. In addition, obesity will also lead to intestinal flora imbalance and intestinal barrier function damage, resulting in the entry of enterogenic endotoxins such as LPS into the blood circulation, activating inflammatory signaling pathways such as NF-κB, and further promoting macrophages and adipocytes in adipose tissue to release a large number of pro-inflammatory factors (such as TNF-α, IL-6, etc.), making the body in a state of low-grade inflammation and aggravating metabolic imbalance and health risks. Therefore, it has important research value to propose a feasible intervention strategy for glucolipid metabolic hormone disorder, oxidative stress state and low-grade inflammation caused by simple obesity.

[0003] As a widely popular natural beverage, black tea is rich in various bioactive components such as tea polyphenols, amino acids and vitamins, showing physiological activities such as antioxidant, lipid-lowering and metabolic regulation. Research shows that the tea polyphenols in black tea can reduce blood lipid levels and improve overall health. "Medicine and food homology" means that while some natural foods provide basic nutrition, they also have the potential to prevent and assist in the treatment of diseases. In recent years, with the in-depth research, medicinal and edible homologous raw materials such as lotus leaf have received attention due to their good lipid-lowering effect, and mulberry and others have been widely studied due to their significant antioxidant activity. On this basis, a traditional Chinese medicine preparation of compound black tea that combines a variety of medicinal and edible homologous ingredients with black tea and plays a positive role simultaneously in regulating glucolipid metabolic hormone disorder, relieving oxidative stress injury and improving low-grade inflammation not only meets the needs of modern consumers for healthy beverages, but also has a large market application space. Summary of the Invention

[0004] Object of the Invention: The first object of the present invention is to provide a traditional Chinese medicine preparation based on black tea and its preparation method. The second object of the present application is to provide the use of the above traditional Chinese medicine preparation in improving simple obesity.

[0005] Technical solution: The present invention provides a traditional Chinese medicine preparation based on black tea, and the traditional Chinese medicine preparation is made from the following components by weight: 10-20 parts of black tea, 5-10 parts of raspberry, 5-10 parts of semen alpiniae oxyphyllae, and 5-10 parts of hawthorn.

[0006] Further, the black tea is Jiuqu Hongmei or Jiaye Long tea.

[0007] The present invention also provides a preparation method of the above traditional Chinese medicine preparation. The black tea, raspberry, semen alpiniae oxyphyllae, and hawthorn are respectively pulverized and sieved, and after being mixed evenly, the raw material powder is extracted in warm water. After the extraction is completed, the extract is centrifuged, and the supernatant is taken.

[0008] Further, the extraction conditions are extraction at 60-70 °C for 2-3 h; the centrifugation conditions are centrifugation at 5000-6000 r / min for 5-10 min.

[0009] Further, the solid-liquid ratio of the raw material powder to warm water is 5-20 mg / mL.

[0010] The present invention also provides the use of the above traditional Chinese medicine preparation in improving simple obesity.

[0011] Further, the multi-faceted improvement of simple obesity includes alleviating problems related to glycolipid metabolism, oxidative stress injury, and low-grade inflammatory state caused by simple obesity.

[0012] Further, the problems related to glycolipid metabolism include disorders of glycolipid metabolism hormones, abnormal activities of glycolipid metabolism-related enzymes, and glycolipid metabolism levels.

[0013] Further, alleviating disorders of glycolipid metabolism hormones includes reducing the contents of insulin (INS), leptin (LEP), and glucagon-like peptide 1 (GLP-1) in serum and increasing the content of adiponectin (ADPN) in serum; alleviating abnormal activities of glycolipid metabolism-related enzymes includes reducing the activity of pancreatic lipase (PNLIP) in the intestine and increasing the activities of trypsin (TPS), α-amylase (AMY), and α-glucosidase (α-GCS) in the intestine; alleviating glycolipid metabolism levels includes reducing the levels of low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), total cholesterol (TC), and glucose (GLU) in blood and epididymal fat index and increasing the level of high-density lipoprotein cholesterol (HDL-C).

[0014] Further, alleviating oxidative stress injury includes reducing the content of MDA in the liver and increasing the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and the content of glutathione (GSH) in the liver.

[0015] Furthermore, alleviating the low-grade inflammatory state includes reducing the contents of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and lipopolysaccharide (LPS) in the serum.

[0016] Traditional Chinese medicine theory:

[0017] Black tea is warm in nature, sweet and bitter in taste, enters the spleen, stomach, and heart meridians, and has the effects of warming the middle-jiao to dispel cold, promoting diuresis and defecation, and promoting fluid production to relieve vexation. Hawthorn is sour, sweet, and slightly warm in nature, enters the spleen, stomach, and liver meridians, and is good at promoting digestion and resolving food stagnation, and promoting blood circulation to remove blood stasis. Raspberry is sweet, sour, and slightly warm in nature, belongs to the liver and kidney meridians, and has the effects of astringing essence and reducing urination, and tonifying the kidney and nourishing the liver. Alpinia oxyphylla is pungent and warm in nature, belongs to the spleen and kidney meridians, and has the effects of warming and tonifying the spleen and kidney, astringing essence and reducing urination, and warming the stomach to stop diarrhea. When the four are used together, hawthorn mainly focuses on "promoting circulation", raspberry mainly focuses on "tonifying", Alpinia oxyphylla mainly focuses on "warming", and black tea mainly focuses on "harmonizing", jointly constituting a compound compatibility system of "combining promoting circulation and tonifying, warming yang and promoting blood circulation, regulating the middle-jiao and harmonizing the stomach", and has a good overall conditioning effect in regulating the functions of zang-fu organs, improving the metabolic state, and alleviating the endogenous production of deficiency heat.

[0018] Basis of modern food function theory:

[0019] Black tea contains theaflavins, thearubigins, tea polysaccharides, caffeine, etc., and has the effects of antioxidant and promoting fat decomposition. Hawthorn is rich in polyphenols, triterpenoids, pectin and other components, and has good functions of reducing blood lipid, promoting digestion, and improving cholesterol metabolism, and is especially suitable for controlling weight, assisting in regulating blood lipid and blood sugar. Raspberry contains rich functional components such as anthocyanins, ellagic acid, and dietary fiber, which helps to resist insulin resistance and regulate lipid metabolism. Alpinia oxyphylla is rich in volatile oils, flavonoids, sesquiterpenoids and other components, and can promote the absorption and transformation of nutrients. The four complement each other in terms of active ingredients, physiological functions and action mechanisms, forming a composite function combination with "lipid-lowering + antioxidant + anti-inflammatory" as the core, with significant scientific compatibility rationality, and is suitable for developing into a health food with multiple effects such as metabolic regulation, anti-inflammatory and antioxidant.

[0020] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: The traditional Chinese medicine preparation based on black tea disclosed in this application not only shows good alleviation of glycolipid metabolic hormone disorders, but also can effectively alleviate the oxidative stress damage of the body and the low-grade inflammatory state related to obesity, and has good potential application value. Brief description of the drawings

[0021] Figure 1 Graph showing the changes in the levels of glycolipid metabolic hormone disorders in rats of different groups. # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents the comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001.

[0022] Figure 2 Graph showing the changes in the activities of glycolipid metabolism-related enzymes in rats of different groups. # represents comparison between the model group and the blank group, # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001.

[0023] Figure 3 Graph showing the changes in glycolipid metabolism levels in rats of different groups. # represents comparison between the model group and the blank group, # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001.

[0024] Figure 4 Graph showing the changes in the activities of liver antioxidant-related enzymes, and the levels of malondialdehyde and glutathione in rats of different groups. # represents comparison between the model group and the blank group, # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001.

[0025] Figure 5 Graph showing the changes in the low-grade inflammation levels in the bodies of rats of different groups. # represents comparison between the model group and the blank group, # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001.

[0026] Figure 6 Graph showing the changes in the body weights of rats of different groups and the body weights of rats in the last week. # represents comparison between the model group and the blank group, # represents P < 0.05, ## represents P < 0.01, represents P < 0.001; * represents comparison between the experimental group and the model group, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001. Detailed implementation manner

[0027] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings.

[0028] Example 1: Screening of black tea and varieties of medicated and edible homologous substances

[0029] 1. Selection and processing methods of black tea and varieties of medicated and edible homologous substances

[0030] The black tea varieties are: Pu'er, Dianhong, Phoenix Dancong, Jiuqu Hongmei, Jiaye Longcha, Keemun Black Tea, Anji Dark Tea, and Jinjunmei; the varieties of medicine and food homology are: Mesona chinensis, Hawthorn, Poria cocos, Coix seed, Raspberry, Mulberry leaf, Alpinia oxyphylla, Kudzu root, Tangerine peel, Lycium barbarum, Mesona chinensis, Adzuki bean, Cassia seed, Gardenia jasminoides, and Chrysanthemum morifolium.

[0031] Each black tea and each medicine and food homology were separately pulverized and passed through a 100-mesh sieve to obtain black tea and fine powders of different varieties of medicine and food homology. According to the solid-liquid ratio of 1:100 g / mL, each raw material fine powder was extracted in warm water at 65°C for 2 h, centrifuged at 5000 r / min for 5 min, and the supernatant was taken to obtain black tea extract and extract of medicine and food homology, which were refrigerated at 4°C for standby.

[0032] 2. Screening of black tea and medicine and food homology varieties

[0033] (1) Determination of in vitro enzyme activity inhibition rate

[0034] S1. Determination of the in vitro activity inhibition rate of α-glucosidase

[0035] In 23 96-well plates, 50 μL of black tea extract or extract of different varieties of medicine and food homology and 50 μL of 3.38 U / mL α-glucosidase solution were added respectively. After incubation at 37°C for 10 min, 50 μL of 2 mg / mL 4-nitrophenyl-α-D-glucopyranoside (pNPG) solution was added, and incubation was continued at 37°C for 20 min. Then, 80 μL of 0.2 mol / L Na2CO3 was added to the mixture to terminate the reaction, and the absorbance was measured at 405 nm. The α-glucosidase activity inhibition rate was calculated according to the following formula:

[0036] α-glucosidase inhibition rate / % = [1 - (a1 - a2) / (a3 - a4)] × 100

[0037] Where: a1 is the absorbance value of the common reaction of the sample (black tea extract or extract of medicine and food homology), α-glucosidase, and pNPG; a2 is the absorbance value of the common reaction of the sample (black tea extract or extract of medicine and food homology), α-glucosidase, and deionized water; a3 is the absorbance value of the common reaction of deionized water, α-glucosidase, and pNPG; a4 is the absorbance value of the common reaction of deionized water and α-glucosidase.

[0038] S2. Determination of the in vitro activity inhibition rate of pancreatic lipase

[0039] Add 50 μL of the extraction solutions of different varieties of black tea or different varieties of extracts of medicinal and edible homologous substances and 50 μL of 1 mg / mL pancreatic lipase solution into 23 96-well plates respectively. After incubation at 37 °C for 10 min, add 50 μL of 2 mg / mL 4-nitrophenyl butyrate (pNPB) solution. After continuing incubation at 37 °C for 20 min, measure the absorbance at 405 nm. The inhibition rate of pancreatic lipase activity is calculated according to the following formula:

[0040] Inhibition rate of pancreatic lipase / % = [1 - (a5 - a6) / (a7 - a8)] × 100

[0041] Where: a5 is the absorbance value of the common reaction of the sample (black tea extraction solution or extract of medicinal and edible homologous substances), pancreatic lipase and pNPB; a6 is the absorbance value of the common reaction of the sample (black tea extraction solution or extract of medicinal and edible homologous substances), deionized water and pNPB; a7 is the absorbance value of the common reaction of adding deionized water, pancreatic lipase and pNPB; a8 is the absorbance value of the common reaction of adding deionized water and pNPB.

[0042] S3. Determination of the in vitro activity inhibition rate of cholesterol esterase

[0043] Dissolve pNPB in a phosphate buffer solution with a pH value of 7.0. Add 50 μL of the extraction solutions of different varieties of black tea or different varieties of extracts of medicinal and edible homologous substances and 50 μL of 0.663 mg / mL cholesterol esterase solution into 23 96-well plates respectively. After incubation at 37 °C for 10 min, add 50 μL of 2 mg / mL 4-nitrophenyl butyrate (pNPB) solution. After continuing incubation at 37 °C for 20 min, measure the absorbance at 405 nm. The inhibition rate of cholesterol esterase activity is calculated according to the following formula:

[0044] Inhibition rate of cholesterol esterase / % = [1 - (a9 - a 10 ) / (a 11 - a 12 )] × 100

[0045] Where: a9 is the absorbance value of the common reaction of the sample (black tea extraction solution or extract of medicinal and edible homologous substances), cholesterol esterase and pNPB, a 10 is the absorbance value of the common reaction of the sample (black tea extraction solution or extract of medicinal and edible homologous substances), buffer solution and pNPB, a 11 is the absorbance value of the common reaction of deionized water, cholesterol esterase and pNPB, a 12 is the absorbance value of the common reaction of buffer solution, deionized water and pNPB.

[0046] (2) Determination of in vitro antioxidant activity

[0047] S1. Scavenging rate of DPPH free radicals

[0048] The determination of DPPH free radical scavenging ability was carried out according to the instructions of the kit (Nanjing Jiancheng Bioengineering Institute, product number: A153-1-1). According to the instructions, prepare DPPH ethanol solution as the working solution. Pipette 400 μL of the sample to be tested (tea extracts of different varieties or extracts of different varieties of medicinal and edible homologous substances) and 600 μL of the working solution, vortex and mix well. After reacting in the dark for 30 min, measure the absorbance at 517 nm. Use 80% methanol to replace the sample and the working solution respectively as the blank hole and the control hole. The DPPH free radical scavenging rate was calculated according to the following formula:

[0049] DPPH free radical scavenging ability / % = [1 - (a 13 - a 14 ) / a 15 × 100

[0050] In the formula: a 13 is the absorbance value of adding the sample and DPPH working solution, a 14 is the absorbance value of adding the sample and 80% methanol solution, a 15 is the absorbance value of adding 80% methanol and DPPH working solution.

[0051] Draw a standard curve with 5 - 25 μg / mL Trolox standard application solution, and the results are expressed as the equivalent amount of Trolox per milliliter (μg Trolox / mL).

[0052] S2. Scavenging rate of ABTS free radicals

[0053] The determination of total antioxidant capacity (ABTS method) was carried out according to the instructions of the kit (Nanjing Jiancheng Bioengineering Institute, product number: A015-2-1). According to the instructions, prepare ABTS working solution. Pipette 10 μL of the sample to be tested (tea extracts of different varieties or extracts of different varieties of medicinal and edible homologous substances) and 190 μL of the working solution, vortex and mix well. After reacting at room temperature for 6 min, measure the absorbance at 405 nm. Use distilled water to replace the sample and the working solution respectively as the blank hole and the control hole. Draw a standard curve with 0.1 - 1.0 mM Trolox standard application solution, and the results are expressed as the equivalent amount of Trolox per milliliter (mM TEAC).

[0054] S3. Scavenging rate of Fe 2+ free radicals

[0055] The determination of the total antioxidant capacity (FRAP method) was carried out according to the instructions of the kit (Nanjing Jiancheng Bioengineering Institute, catalog number: A015-3-1). According to the requirements of the instructions, the FRAP working solution was prepared. 5 μL of the sample to be tested (black tea extract or the extract of medicinal and edible homologous substances) was vortexed and mixed evenly with 180 μL of the working solution. After reacting at 37 °C for 5 min, the absorbance was measured at 593 nm. Distilled water was used instead of the sample as the blank hole, and a standard curve was plotted with a 0.15 - 1.5 mM FeSO4·7H2O solution. The results were expressed as the equivalent of FeSO4 per milliliter (mM FeSO4).

[0056] (3) Comparison of antioxidant and enzyme activity inhibition rates of extracts

[0057] The comprehensive scores of the antioxidant and enzyme activity inhibition rates of all black tea extracts and extracts of medicinal and edible homologous substances were calculated by the comprehensive evaluation normalization method. The top two black tea varieties and the top three varieties of medicinal and edible homologous substances with the highest comprehensive scores were selected for subsequent experiments. The calculation method of the comprehensive score is as follows. The calculation results of black tea are shown in Table 1, and the calculation results of medicinal and edible homologous substances are shown in Table 2:

[0058] Comprehensive score / point = [(b i / b max +c i / c max +d i / d max +e i / e max +f i / f max +g i / g max ) / 6]×100

[0059] Where: b i , c i , d i , e i , f i and g i correspond to the α-glucosidase activity inhibition rate, pancreatic lipase activity inhibition rate, cholesterol esterase activity inhibition rate, DPPH free radical scavenging ability, total antioxidant capacity (ABTS method), and total antioxidant capacity (FRAP method) of the extract, respectively; b max , c max , d max , e max , f max and g max correspond to the highest values among the groups for the inhibition rates of the three glycolipid metabolism enzymes and the three antioxidant capacity measurement methods, respectively.

[0060] Table 1 Comprehensive score table of lipid-lowering and antioxidant effects of black tea

[0061]

[0062]

[0063] Table 2 Comprehensive score table of lipid-lowering and antioxidant effects of homologous substances of medicine and food

[0064]

[0065]

[0066] Note: Different letters indicate significant differences between samples (P<0.05)

[0067] Example 2: Determination of traditional Chinese medicine preparation formula

[0068] Select the best-performing black tea Jiupu Hongmei in Example 1, combine it with different homologous substances of medicine and food respectively, prepare the extracts of different formulas according to the method in Example 1, and detect the enzyme activity inhibition ability and in vitro antioxidant ability. The specific formula information is shown in Table 3. The results are shown in Table 4.

[0069] Table 3 Traditional Chinese medicine preparation formula table

[0070]

[0071]

[0072] Table 4 Comprehensive score table of lipid-lowering and antioxidant effects of traditional Chinese medicine preparations

[0073]

[0074] Note: Different letters indicate significant differences between samples (P<0.05)

[0075] According to the results in Table 4, select Jiupu Hongmei: Rubus idaeus: Alpinia oxyphylla: Crataegus pinnatifida = 1:0.5:0.5:0.5 as the optimal formula for subsequent animal experiments.

[0076] Example 3 Animal experiment verification of the efficacy of traditional Chinese medicine preparations

[0077] 1. Animal grouping and establishment of obese rat models

[0078] Select 49 4-week-old SPF-grade Waster male rats (Jinan Pengyue Laboratory Animal Breeding Co., Ltd., license number: SCXK(Lu)20220006, 120±20g). The laboratory adopts 12h light and dark rotation lighting, has good ventilation and sanitary conditions, the room temperature is 22±2°C, and the humidity is 60±5%. Before the experiment, all rats were fed with ordinary feed for 1 week for environmental adaptation. During this period, the rats could freely eat ordinary feed and drink water.

[0079] Then, 49 rats were randomly divided into 7 groups, with no significant difference in average body weight among groups (P>0.05). The blank group (NC) continued to be fed with normal feed (20% flour, 10% rice flour, 20% corn, 26% bran, 20% bean material, 2% fish meal, 2% bone meal), and the model group (HFD), positive control group (PC), experimental group 1 (MJQHM), experimental group 2 (MJYL), experimental group 3 (LJQHM), and experimental group 4 (HJQHM) were fed with high-fat feed (78.80% normal feed, 10% lard, 10% egg yolk powder, 1% cholesterol, 0.2% bile salt). By changing the solid-liquid ratio of the extraction of traditional Chinese medicine preparation 1 (Jiuxu Hongmei + Hawthorn + Raspberry + Alpinia oxyphylla = 1:0.5:0.5:0.5) (i.e., 1:50, 1:100, 1:200), leaching solutions of traditional Chinese medicine preparation 1 with different extraction concentrations (20 mg / mL, 10 mg / mL, 5 mg / mL) were prepared for animal experiments. After 4 weeks of modeling, the PC group was intragastrically administered orlistat solution, the LJQHM group, MJQHM group, and HJQHM group were intragastrically administered traditional Chinese medicine preparation 1 solutions with different concentrations, the MJYL group was intragastrically administered traditional Chinese medicine preparation 2 (Jiaye Longcha + Hawthorn + Raspberry + Alpinia oxyphylla = 1:0.5:0.5:0.5) solution, and the NC group and HFD group were intragastrically administered normal saline. The intervention time was 4 weeks. The grouping and treatment conditions are shown in Table 5.

[0080] Table 5 Grouping of Animal Experiments

[0081]

[0082] 2. Determination of Experimental Indexes

[0083] After the intervention experiment ended, all rats were fasted and water-deprived for 12 h. Then, the following indexes (1)-(5) were measured.

[0084] (1) Levels of Glucolipid Metabolism Hormones

[0085] After anesthesia with ether, blood was immediately collected by puncturing the eyeball. The collected blood was incubated in a 37 °C incubator for 30 min, centrifuged at 3500 r / min at 4 °C for 10 min to obtain serum. The contents of ADPN, LEP, INS, and GLP-1 in the serum were measured using ADPN, LEP, INS, and GLP-1 ELISA kits (Shanghai Hualan Chemical Technology Co., Ltd.).

[0086] (2) Levels of Glucolipid Metabolism-Related Enzyme Activities

[0087] After decapitating the rats to sacrifice them, collect the small intestine tissues of the rats, remove the contents, add PBS solution at a mass-to-volume ratio of 1:9 (g / mL), grind well in an ice bath, transfer to a 10 mL centrifuge tube, centrifuge at 10000 r / min for 10 min, and take the supernatant for enzyme activity determination. Determine the activities of PNLIP, TPS, AMY, and α-GCS in the intestine using PNLIP, TPS, AMY, and α-GCS enzyme activity assay kits (Nanjing Jiancheng Bioengineering Institute, catalog numbers: A054-1-1, A080-2, C016-1-1, and H419-1-2).

[0088] (3) Glycolipid metabolism level

[0089] Collect serum and analyze the contents of TC, TG, LDL-C, HDL-C, and GLU in the serum using a Hitachi 7020 fully automatic biochemical analyzer. After decapitating the rats to sacrifice them, collect the epididymal fat of the rats, weigh it, and calculate the epididymal fat index by dividing the epididymal fat weight by the rat body weight.

[0090] (4) Oxidative stress injury level

[0091] After decapitating the rats to sacrifice them, collect the liver, take 1.0 g of the liver and put it into a homogenizer, add 9 mL of PBS solution, grind well in an ice bath to homogenize the liver tissue, transfer to a 10 mL centrifuge tube, centrifuge at 3000 r / min for 15 min, take the supernatant of the homogenate after centrifugation and transfer it to a new 10 mL centrifuge tube, and store it at 4 °C for later use. Take the supernatant of the liver homogenate and determine the activities of SOD and GSH-Px, as well as the contents of GSH and MDA in the liver homogenate using SOD enzyme activity, GSH-Px enzyme activity, GSH, and MDA content assay kits (Beijing Solarbio Science & Technology Co., Ltd., catalog numbers: BC0170, BC1195, BC1175, and BC0025).

[0092] (5) Low-grade inflammation level

[0093] Collect serum and determine the contents of NF-κB, TNF-α, IL-6, and LPS in the serum using NF-κB, TNF-α, IL-6, and LPS ELISA kits (Shanghai Hualan Chemical Technology Co., Ltd.).

[0094] (6) Body weight and body weight gain

[0095] Weigh the rats at the end of each week and plot a graph of the rat body weight change trend. Subtract the rat body weight at the start of the intervention from the rat body weight after 4 weeks of intervention to obtain the rat body weight gain.

[0096] 3. Experimental results

[0097] (1) General overview of rat glycolipid metabolism hormone levels

[0098] The levels of hormones related to glycolipid metabolism in rats were as follows Figure 1 As shown, in the HFD group, the ADPN in the serum of rats Figure 1 a) decreased to 547.9 ± 84.92 ng / mL, significantly lower than that in the NC group (P < 0.05), and the INS Figure 1 c), GLP-1 Figure 1 d) and LEP Figure 1 b) reached 55.29 ± 3.15 mU / L, 19.00 ± 1.89 pmol / L and 3.158.39 ± 0.82 ng / mL respectively, significantly higher than those in the NC group (P < 0.05), indicating that the obese state induced by HFD led to disorders of hormones related to glycolipid metabolism. Compared with the HFD group, the contents of ADPN, LEP, INS and GLP-1 in the sera of rats in the LJQHM, MJQHM, HJQHM and MJYL groups intragastrically administrated with the traditional Chinese medicine preparation were all regulated. Among them, the ADPN in the sera of rats in the LJQHM, MJQHM, HJQHM and MJYL groups was significantly higher than that in the HFD group (P < 0.05), and the contents of INS, GLP-1 and LEP in the sera of rats in the MJQHM, HJQHM and MJYL groups were significantly lower than those in the HFD group (P < 0.05). Among them, the MJQHM group had the best regulatory effect, the ADPN content recovered to 1897.39 ± 183.29 ng / mL, and the contents of INS, GLP-1 and LEP recovered to 31.56 ± 2.90 mU / L, 9.01 ± 1.33 pmol / L and 4.80 ± 0.70 ng / mL respectively. The above results indicate that this traditional Chinese medicine preparation can effectively increase the serum ADPN level, decrease the levels of INS, GLP-1 and LEP, and then improve the disorders of hormones related to glycolipid metabolism in the body, and has the efficacy of regulating glycolipid metabolism.

[0099] (2) General situation of the levels of enzymes related to glycolipid metabolism in rats

[0100] The changes in the levels of enzymes related to glycolipid metabolism in rats were as follows Figure 2 As shown, long-term high-fat diet led to a significant increase in the activity of PNLIP Figure 2 a) in the intestine of rats in the HFD group, reaching 728.55 ± 11.32 U / g prot, significantly higher than that in the NC group (P < 0.05), and the TPS Figure 2 b), AMY Figure 2 d), α-GCS Figure 2c) The activities decreased to 9.73±1.36 U / mgprot, 11.04±0.42 U / mgprot, and 552.99±10.64 U / mgprot respectively, all significantly lower than those of the NC group (P<0.05). Compared with the HFD group, after intervention with the traditional Chinese medicine preparation, the activities of PNLIP in the intestines of rats in the LJQHM, MJQHM, HJQHM, and MJYL groups were significantly decreased. Among them, the effect of the MJQHM group was the most significant, reaching 253.83±20.77 U / gprot. In addition, the activities of TPS, AMY, and α-GCS in the intestines of the LJQHM and MJQHM groups were significantly increased compared with those of the HFD group rats (P<0.05), and were significantly higher than those of the MJYL group, reaching 22.84±4.35 U / mgprot, 18.42±0.22 U / mgprot, and 659.98±12.62 U / mgprot. In summary, the traditional Chinese medicine preparation can effectively regulate the activities of glycolipid metabolism enzymes in the intestine, and thus regulate the body's glycolipid metabolism.

[0101] (3) General situation of rat glycolipid metabolism level

[0102] The blood lipid, epididymal fat index, and blood glucose levels of rats were as Figure 3 shown, and the blood lipid level was as Figure 3 shown in a. After feeding with high-fat diet, the contents of TC, TG, and LDL-C in the serum of rats in the HFD group reached 2.65±0.18 mmol / L, 1.65±0.21 mmol / L, and 0.43±0.02 mmol / L respectively, all significantly higher than those of the NC group (P<0.05), and the HDL-C reached 0.58±0.04 mmol / L, significantly lower than that of the NC group (P<0.05), indicating that high-fat diet induced blood lipid metabolism disorder. Compared with the HFD group, the levels of TC and TG in the LJQHM, MJQHM, HJQHM, and MJYL groups after intragastric administration of the traditional Chinese medicine preparation were decreased, and were significantly lower than those of the HFD group (P<0.05), and the levels of HDL-C were increased, significantly higher than those of the HFD group (P<0.05). Among them, the MJQHM group had the best recovery effect, and the levels of TC, TG, and HDL-C were restored to 1.94±0.08 mmol / L, 0.60±0.01 mmol / L, and 0.63±0.03 mmol / L respectively. The epididymal fat index level was as Figure 3 shown in c. The epididymal fat index of rats in the HFD group reached 2.64±0.13 g / g, significantly higher than that of the NC group (P<0.05), indicating that after a long-term high-fat diet, obvious fat accumulation occurred in HFD rats. After intragastric administration of the traditional Chinese medicine preparation, the epididymal fat index of rats was significantly decreased. Among them, the HJQHM group had the best quality effect, and the epididymal fat index decreased to 1.91±0.24 g / g. It shows that this compound functional preparation helps to promote lipid metabolism balance and relieve lipid accumulation.

[0103] The blood glucose level was as follows Figure 3 As shown in b, the GLU in the serum of rats in the HFD group reached 7.62 ± 0.10 mmol / L, which was significantly higher than that in the NC group (P < 0.05), indicating that a high-fat diet led to glucose metabolism disorder in rats. Compared with the HFD group, after the intervention of intragastric administration of the two traditional Chinese medicine preparations, the blood glucose levels in the LJQHM, MJQHM, HJQHM, and MJYL groups were all significantly decreased (P < 0.05). Among them, the MJQHM group had the best therapeutic effect, recovering to 6.98 ± 0.17 mmol / L, which also indicated that the traditional Chinese medicine preparation could effectively relieve the glucose metabolism disorder caused by simple obesity.

[0104] (4) Overview of liver oxidative stress level

[0105] The activities of antioxidant enzymes and the contents of malondialdehyde and glutathione in the livers of rats were as follows Figure 4 As shown, the antioxidant capacity of the livers of rats in the HFD group was significantly decreased (P < 0.05). Among them, the activities of GSH-Px ( Figure 4 a), SOD ( Figure 4 b) were decreased to 0.35 ± 0.01 U / mgprot and 15.74 ± 2.88 U / mg prot respectively, the content of GSH ( Figure 4 d) was decreased to 262.97 ± 11.01 μg / mg, and the content of MDA ( Figure 4 c) was increased to 0.20 ± 0.01 nmol / mgprot, indicating that a high-fat diet induced obvious oxidative stress damage. After intragastric administration of the traditional Chinese medicine preparation, the MJQHM group had a better recovery effect compared with the LJQHM group and the HJQHM group. The activities of GSH-Px, SOD, and the content of GSH were restored to 0.48 ± 0.01 U / mg prot, 62.27 ± 5.58 U / mg prot, and 455.59 ± 44.59 U / mgprot respectively, which were significantly higher than those in the HFD group (P < 0.05). The content of MDA was restored to 0.13 ± 0.01 nmol / mg prot, which was significantly lower than that in the HFD group (P < 0.05). It should be noted that the activities of SOD, GSH-Px and the content of GSH in the MJQHM group were all higher than those in the MJYL group, but the difference was not significant (P > 0.05), indicating that it had certain advantages in antioxidant. In summary, the traditional Chinese medicine preparation could effectively enhance the activities of liver antioxidant enzymes, reduce the MDA level, and relieve the oxidative stress damage induced by a high-fat diet.

[0106] (5) Overview of the inflammatory level of rats

[0107] The inflammatory level of rats was as follows Figure 5 As shown, the NF-κB ( Figure 5 a), LPS ( Figure 5 b), TNF-α (Figure 5 c) and IL-6( Figure 5 d) The levels reached 1520.3 ± 89.25 pg / mL, 843.10 ± 36.75 EU / L, 395.98 ± 17.52 pg / mL, and 52.30 ± 2.17 pg / mL respectively, all significantly higher than those in the NC group (P < 0.05), indicating that the HFD-induced low-grade inflammatory state was exacerbated. However, after the intervention with the traditional Chinese medicine preparation, the inflammatory levels in the rats of the LJQHM, MJQHM, HJQHM, and MJYL groups were all restored to a certain extent, and the MJQHM group had the best recovery effect on the inflammatory level. The levels of NF-κB, LPS, TNF-α, and IL-6 were restored to 502.36 ± 41.44 pg / mL, 115.46 ± 16.76 EU / L, 416.59 ± 37.78 pg / mL, and 37.41 ± 3.48 pg / mL respectively. The above experimental results show that the traditional Chinese medicine preparation can effectively reduce the levels of inflammatory factors, inhibit the systemic inflammatory response induced by HFD, thereby improving the systemic inflammatory environment, alleviating the systemic low-grade inflammatory response, and further verifying its potential role and application value in regulating the low-grade inflammatory state.

[0108] (6) Overview of body weight and body weight gain

[0109] The body weight change of the rats was as Figure 6 shown in a, and the body weight gain was as Figure 6 shown in b. As can be seen from Figure 6 b, after feeding with a high-fat diet, the body weight gain of the rats in the HFD group reached 317.79 ± 20.53 g, significantly higher than that in the NC group (P < 0.05). After the intervention, the body weight gains of the MJQHM and HJQHM groups decreased to 280.49 ± 8.19 g and 271.38 ± 9.97 g respectively, significantly lower than that in the HFD group (P < 0.05). The traditional Chinese medicine preparation has the effect of inhibiting the body weight gain induced by a high-fat diet and can effectively alleviate the obesity caused by a high-fat diet.

Claims

1. A traditional Chinese medicine preparation based on black tea, characterized in that, The traditional Chinese medicine preparation is prepared from the following components in parts by weight: 10-20 parts of black tea, 5-10 parts of raspberry, 5-10 parts of semen alpiniae oxyphyllae, and 5-10 parts of hawthorn.

2. The traditional Chinese medicine preparation based on black tea according to claim 1, characterized in that, The black tea is Jiuqu Hongmei or Jiaye Longcha.

3. The preparation method of the traditional Chinese medicine preparation based on black tea according to claim 1, characterized in that, The black tea, raspberry, semen alpiniae oxyphyllae, and hawthorn are respectively pulverized and sieved, and after being evenly mixed, the raw material powder is extracted in warm water. After the extraction is completed, the extract is centrifuged, and the supernatant is taken.

4. The preparation method according to claim 3, characterized in that, The solid-liquid ratio of the raw material powder to warm water is 5-20 mg / mL.

5. Use of the traditional Chinese medicine preparation according to any one of claims 1 to 2 in improving simple obesity.

6. The use according to claim 5, characterized in that, The improvement of simple obesity includes alleviating problems related to glycolipid metabolism, oxidative stress injury, and low-grade inflammatory state caused by simple obesity.

7. The use according to claim 6, characterized in that, The problems related to glycolipid metabolism include disorders of glycolipid metabolism hormones, abnormal activities of glycolipid metabolism-related enzymes, and glycolipid metabolism levels.

8. The use according to claim 7, wherein Alleviating disorders of glycolipid metabolism hormones includes reducing the contents of insulin, leptin, and glucagon-like peptide 1 in serum and increasing the content of adiponectin in serum; alleviating abnormal activities of glycolipid metabolism-related enzymes includes reducing the activity of lipase in the intestine and increasing the activities of trypsin, α-amylase, and α-glucosidase in the intestine; alleviating glycolipid metabolism levels includes reducing the levels of low-density lipoprotein cholesterol, triglyceride, total cholesterol, and glucose in blood and epididymal fat index and increasing the level of high-density lipoprotein cholesterol.

9. The use according to claim 6, characterized in that, Alleviating oxidative stress injury includes reducing the content of malondialdehyde in the liver and increasing the activities of superoxide dismutase, glutathione peroxidase, and the content of glutathione in the liver.

10. The use according to claim 6, characterized in that, Alleviating low-grade inflammatory state includes reducing the contents of nuclear factor-κB, tumor necrosis factor-α, interleukin-6, and lipopolysaccharide in serum.

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