Application of codonopsis pilosula aqueous extract in preparation of product for relieving or treating food-borne obesity

Abstract

The invention relates to the technical field of traditional Chinese medicine extracts, in particular to application of a codonopsis pilosula aqueous extract in preparation of a product for relieving or treating food-borne obesity. It is found for the first time through experimental research that the codonopsis pilosula aqueous extract can effectively improve the leptin level in hypothalamus of a food-borne obese mouse, inhibit expression of neurons NPY / Agrp for promoting ingestion and relieve central leptin resistance, so that appetite is reduced, food intake is reduced, then body weight is reduced, lipid deposition in the liver is reduced, and the weight of the mouse is reduced. Therefore, the composition can be used for preparing products for relieving or treating the food-borne obesity, such as functional foods beneficial to controlling fat in a body or medicines for treating the food-borne obesity or relieving symptoms of the food-borne obesity. The composition can be used for preparing the products for relieving or treating the food-borne obesity, such as the functional foods beneficial to controlling fat in the body or the medicines for treating the food-borne obesity or relieving symptoms of the food-borne obesity.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine extracts, and in particular to application of a water extract of Codonopsis pilosula in preparing a product for alleviating or treating food-induced obesity. Background Art

[0002] With the improvement of living conditions, the fat energy supply ratio in the diet of Chinese residents has continued to increase, resulting in a rapid increase in the incidence of food-induced obesity across the country. Food-induced obesity is the result of excessive fat accumulation when the body's energy intake is greater than its energy consumption for a long time. It is the core link in causing a variety of metabolic diseases such as type 2 diabetes, fatty liver, and cancer, and it seriously threatens human health. In addition, according to the 2023 World Obesity Federation (WOF) report analysis, the number of obese patients is continuing to grow rapidly, and the prevalence of food-induced obesity is showing an increasing trend year by year. It is estimated that by 2035, 51% of the world's population will face obesity or overweight problems, involving a total population of more than 4 billion. Therefore, preventing the occurrence and development of food-induced obesity has become a clinical problem that urgently needs to be solved.

[0003] Leptin is secreted by adipose tissue and is one of the important regulatory factors responsible for regulating eating behavior and energy metabolism. Under normal physiological conditions, leptin can reach the central nervous system through the blood-brain barrier and transmit satiety signals to the hypothalamus, thereby suppressing appetite and increasing energy consumption. In addition, leptin can also act on the periphery to regulate glucose and lipid metabolism. However, obese patients have too many white fat cells in their bodies, and white fat secretes excessive leptin. The high level of circulating leptin not only causes a decrease in peripheral leptin sensitivity, leading to abnormal glucose and lipid metabolism, but also reduces central leptin sensitivity, so that the hypothalamus cannot perceive the satiety signal in time to stop eating, ultimately leading to overeating and reduced metabolism. This phenomenon is leptin resistance. At present, leptin resistance is a huge challenge facing obesity treatment in my country and even the world. Summary of the invention

[0004] In view of the above technical problems, the present invention provides the use of Codonopsis pilosula water extract in the preparation of a product for treating food-induced obesity. The present invention has found through research that Codonopsis pilosula water extract can increase the level of leptin in the hypothalamus, reduce central leptin resistance, and restore leptin sensitivity, thereby reducing appetite and weight, and effectively alleviating and treating food-induced obesity.

[0005] In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solutions: The first aspect of the present invention provides the use of an aqueous extract of Codonopsis pilosula in preparing a product for alleviating or treating food-induced obesity.

[0006] Codonopsis pilosula is a traditional Chinese medicine that is both a medicine and a food. It is mainly derived from the dried roots of Codonopsis pilosula, Codonopsis pilosula or Codonopsis pilosula of the Campanulaceae family. It is mild in nature and sweet in taste. It has the effects of tonifying the middle, replenishing qi, and promoting the production of body fluids. It is mainly used to treat spleen and stomach weakness, deficiency of qi and blood, fatigue, thirst, etc. Since Codonopsis pilosula can replenish the middle and replenish qi, enhance spleen and stomach function, and promote gastrointestinal motility, it is often used to improve symptoms such as loss of appetite. The present invention unexpectedly discovered through experimental research that for food-induced obesity, Codonopsis pilosula water extract can increase the level of leptin in the hypothalamus, reduce central leptin resistance, and enhance the inhibitory effect of leptin on neurons that promote feeding such as Agrp / NPY, thereby reducing appetite, reducing food intake, and then reducing body weight and reducing lipid deposition in the liver, achieving the effect of alleviating and treating food-induced obesity.

[0007] Preferably, the preparation method of the Codonopsis pilosula water extract is: crush the Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

[0008] More preferably, the Codonopsis pilosula is crushed to a particle size of less than 2 mm.

[0009] More preferably, the heating temperature is 95-105°C.

[0010] More preferably, before the water heating extraction, the crushed Codonopsis pilosula is soaked in water for 1 hour.

[0011] Further preferably, the extraction is performed twice by heating with water and the extracts are combined; during each extraction, the mass ratio of party ginseng to water is 1:8-12, and further preferably is 1:10.

[0012] More preferably, the drying temperature is 45-55°C.

[0013] A second aspect of the present invention provides a functional food that helps control body fat, wherein the active ingredient of the functional food comprises an aqueous extract of Codonopsis pilosula.

[0014] Preferably, the dosage form of the functional food includes but is not limited to tablets, powders, granules, capsules, tea bags and the like.

[0015] Preferably, the preparation method of the Codonopsis pilosula water extract is: crush the Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

[0016] More preferably, the Codonopsis pilosula is crushed to a particle size of less than 2 mm.

[0017] More preferably, the heating temperature is 95-105°C.

[0018] More preferably, before the water heating extraction, the crushed Codonopsis pilosula is soaked in water for 1 hour.

[0019] Further preferably, the extraction is performed twice by heating with water and the extracts are combined; during each extraction, the mass ratio of party ginseng to water is 1:8-12, and further preferably is 1:10.

[0020] More preferably, the drying temperature is 45-55°C.

[0021] The third aspect of the present invention provides a medicine for treating food-induced obesity or alleviating the symptoms of food-induced obesity, wherein the active ingredient of the medicine comprises an aqueous extract of Codonopsis pilosula.

[0022] Preferably, the drug is a drug for reducing leptin resistance.

[0023] Preferably, the medicine is an oral preparation.

[0024] Preferably, the dosage form of the oral preparation includes tablets, granules, capsules, powders, powders and oral solutions.

[0025] Preferably, the preparation method of the Codonopsis pilosula water extract is: crush the Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

[0026] More preferably, the Codonopsis pilosula is crushed to a particle size of less than 2 mm.

[0027] More preferably, the heating temperature is 95-105°C, and more preferably 100°C.

[0028] More preferably, before the water heating extraction, the crushed Codonopsis pilosula is soaked in water for 1 hour.

[0029] Further preferably, the extraction is performed twice by heating with water and the extracts are combined; during each extraction, the mass ratio of party ginseng to water is 1:8-12, and further preferably is 1:10.

[0030] More preferably, the drying temperature is 45-55°C.

[0031] The beneficial effects of the present invention are as follows: the present invention first discovered that the water extract of Codonopsis pilosula can increase the level of leptin in the hypothalamus of mice with food-induced obesity, and the increase in the level of leptin in the hypothalamus can inhibit the expression of the neurons NPY / Agrp that promote food intake, thereby reducing appetite and reducing food intake, thereby producing the effects of reducing body weight and reducing lipid deposition in the liver, thereby achieving the effect of alleviating and treating food-induced obesity. The present invention experimentally proves that after the water extract of Codonopsis pilosula is given to mice with food-induced obesity model, the level of leptin in the hypothalamus is upregulated, the mRNA expression levels of Agrp and NPY are significantly reduced, the food intake of mice is significantly reduced, the speed of weight gain is slowed down, and the number of fat droplets in the liver is significantly reduced. The present invention provides a new direction and theoretical support for treating food-induced obesity and expanding the scope of application of Codonopsis pilosula. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 is a high-resolution HPLC-MS image of the water extract of Codonopsis pilosula used in the examples of the present invention; Figure 2 This is a graph showing changes in the body weight of mice in Example 1 of the present invention; Figure 3 This is a graph showing the weight changes of mice after three weeks of administration in Example 1 of the present invention ( *p <0.05, **p <0.01, ***p <0.001); Figure 4 is the change in daily food intake after administration of Codonopsis pilosula in Example 1 of the present invention; Figure 5 is the change in the average daily food intake of mice in Example 1 of the present invention ( *p <0.05, **p <0.01); Figure 6 This is an H&E staining image of mouse liver in Example 1 of the present invention; Figure 7 is the expression level of leptin in the serum of mice in Example 1 of the present invention ( *p <0.05, **p <0.01); Figure 8 is the relative expression of leptin in the hypothalamus of mice in Example 1 of the present invention ( *p <0.05); Fig. 9 is the relative expression level of Agrp mRNA in the mouse hypothalamus in Example 1 of the present invention ( *p <0.05, **p <0.01); Fig.10 is the relative expression level of NPY mRNA in the mouse hypothalamus in Example 1 of the present invention ( *p <0.05, **p <0.01). DETAILED DESCRIPTION

[0033] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work fall within the scope of protection of the present invention.

[0034] Patients with diet-induced obesity develop leptin resistance because their fat cells secrete large amounts of leptin, which makes their appetite even stronger, making obesity treatment more difficult.

[0035] The present invention first discovered through experimental research that the water extract of Codonopsis pilosula can effectively increase the level of leptin in the hypothalamus, inhibit the expression of NPY / Agrp, a neuron that promotes food intake, and reduce leptin resistance, thereby reducing appetite, reducing food intake, and further reducing body weight and lipid deposition in the liver, achieving the effect of alleviating and treating food-induced obesity. Therefore, it can be used to prepare a product for alleviating or treating food-induced obesity. Based on the above research results, the embodiment of the present invention provides the use of the water extract of Codonopsis pilosula in preparing a product for alleviating or treating food-induced obesity.

[0036] The embodiment of the present invention also provides a functional food that helps control body fat, and the active ingredient of the functional food includes Codonopsis pilosula water extract.

[0037] The embodiment of the present invention also provides a medicine for treating food-induced obesity or alleviating the symptoms of food-induced obesity, wherein the active ingredient of the medicine comprises water extract of Codonopsis pilosula.

[0038] The following is further described with reference to specific embodiments.

[0039] The reagents, drugs and feeds used in the following examples are shown in Table 1.

[0040] Table 1 Main reagents, drugs and feeds name source Model / Batch No. Codonopsis pilosula Huiyuan Chinese Medicinal Herbs Planting Professional Cooperative, Miaoqian Town, Xia County, Pingshun County, Shanxi Province Orlistat Capsules Watson Pharmaceuticals National Medicine Approval No. H20103180 H&E Kit Solebao G1120 PBS Solebao P1010 Tribromoethanol Sigma-Aldrich T48402 OTC embedding medium SAKURA 4583 4% tissue cell fixative Solebao P1110 sucrose Solebao S8271 Standard feed Hua Fukang 1025 60% high-fat feed Xiaoshu Youtai D12492 The Codonopsis pilosula water extract used in the following examples was prepared by the following method: (1) Crushing: crush the Codonopsis pilosula slices and collect the powder after passing through a 10-mesh sieve; (2) Extraction: Take a 1000 mL flask and weigh 50 g of Codonopsis pilosula powder. The solid-liquid ratio is 1:10, add 500 mL of pure water, soak for 1 hour, heat and extract for 1 hour, filter with filter paper, collect the filtrate, add 500 mL of pure water to the residue again, heat and extract for 1 hour, combine the two extracts to obtain the Codonopsis pilosula water extract, and store at 4°C. Repeat 9 times to extract a total of 450 g of Codonopsis pilosula; (3) Concentration: Place the water extract in a rotary evaporator and evaporate the excess solvent to obtain about 260 mL of Codonopsis pilosula water extract (concentrated solution), then place it in an evaporating dish to continue evaporating excess water to obtain Codonopsis pilosula concentrated solution and store it at 4°C; (4) Drying: The mixture is dried by vacuum drying (50°C) to obtain a dry powder of the Codonopsis pilosula water extract.

[0041] The components and contents of the prepared Codonopsis pilosula water extract are shown in Table 2. Figure 1 As shown: Table 2 Components and contents in water extract of Codonopsis pilosula

[0042] Unless otherwise specified, the raw materials, reagents, drugs or instruments used in the following examples are all conventional commercial products obtained from commercial channels. The methods used in the following examples are all conventional methods in the art unless otherwise specified.

[0043] Example 1 This example investigates the therapeutic effect of Codonopsis pilosula water extract on diet-induced obese mice.

[0044] 1 Experimental animals C57BL / 6J male mice aged 6-8 weeks were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. SCX (Beijing) 2012-0001.

[0045] 2 Experimental methods 2.1 Animal modeling and drug administration (1) Construction of DIO mouse model: Male C57BL / 6J mice were randomly divided into four groups: control group (Chow), high-fat diet group (HFD), Codonopsis pilosula group (DS) and positive drug orlistat group (Orlistat), with 10 mice in each group. The control group was fed with standard feed for 12 weeks; the high-fat diet group and the drug group were fed with 60% high-fat diet for 12 weeks, and the diet-induced obesity mouse model was successfully established.

[0046] (2) Administration: After modeling, each group was given drugs by gavage for 3 weeks. The dosage of the DS group was 8 g / kg (in terms of crude drug), and the dosage of the Orlistat group was 15.6 mg / kg. The Chow group and the HFD group were given an equal amount of pure water. All experiments were reviewed and approved by the Animal Center of Tianjin University of Traditional Chinese Medicine.

[0047] 2.2 Recording of mouse body weight and food intake Observe the activity status of the mice, record the food intake of the mice once a day, record the weight of the mice once a week, and draw a graph of food intake and weight changes.

[0048] 2.3 Mouse tissue collection (1) Blood was collected from the inner canthus of the eye and placed in a 1.5 ml EP tube. The blood sample was allowed to stand for half an hour before centrifugation at 4°C, 6000 rpm, and 20 min. The upper layer of serum was collected and stored in a -80°C refrigerator. After blood collection, the mouse was killed by dislocating the neck, exposing its chest and abdominal cavities, and the liver, heart, spleen, lung, and kidney organs of the mouse were removed. The weight of each tissue of the mouse was weighed.

[0049] (1) After 15 weeks (i.e., 12 weeks of modeling and 3 weeks of oral administration), 5 mice were randomly selected for death by cervical dislocation, and the complete brain tissue was removed. According to the mouse brain atlas, the hypothalamus was removed using a mouse brain slice mold, and the specimen was stored in a -80℃ refrigerator for RT-qPCR, Western blot, ELISA and other experiments.

[0050] (2) The remaining 5 mice in each group were perfused with 4% paraformaldehyde, and the brain tissue was fixed with 4% paraformaldehyde for 12 h, then dehydrated with 10% and 30% sucrose gradients, and finally stored in 30% sucrose solution for immunofluorescence experiments. The liver, heart, spleen, lung, kidney, brown fat (scapular), white fat (epididymis) and muscle were fixed in 4% paraformaldehyde and subsequently used for pathological experiments.

[0051] 2.4 Tissue embedding (1) Fixation: Place the cut tissue blocks in 4% paraformaldehyde for fixation, where the volume ratio of tissue blocks to 4% paraformaldehyde is 1:7.

[0052] (2) Dehydration: 30% ethanol → 50% ethanol → 70% ethanol (2 h on the day of the experiment) → 95% ethanol 1 → 95% ethanol 2 → anhydrous ethanol.

[0053] (3) Transparency: 50% ethanol + 50% xylene → xylene 1 → xylene 2 → xylene 3. During the process of transparency, observe the morphology of the tissue frequently and adjust the transparency time in time. Then observe whether the tissue is transparent under light. After transparency is completed, put it in a small plastic box and mark it.

[0054] (4) Wax dipping: Melt the paraffin wax and keep the temperature at about 55°C for three times. The first wax dipping takes 15 minutes, and the next two times can be extended to 1 hour.

[0055] (5) Embedding: Place the tissue block in a mold filled with wax liquid, then fix the tissue with tweezers. The tissue section should be parallel to the bottom. The embedding process should be quick. Then move it to a cooling table at -10℃, let the paraffin solidify, mark it, and remove the iron trough after 30 minutes. 2.5 H&E staining (1) After fixation, liver tissue was embedded in paraffin and sliced ​​at 5 μm.

[0056] (2) Dewax the sections in xylene twice, 10 min each time. Rehydrate in gradient ethanol (100%, 95%, 85%, 75%), 3 min each time. Soak in distilled water for 2 min.

[0057] (3) Stain with hematoxylin for 4 min and wash away loose color with distilled water.

[0058] (4) Differentiate in differentiation solution for 20 s and rinse twice with tap water for 3-5 min each time.

[0059] (5) Stain the sample in eosin solution for 2 min and then wash off the excess solution with tap water.

[0060] (6) Dehydration, transparency, and sealing: Rapid dehydration is performed in the order of ethanol (75%, 85%, 95%, 100%) and xylene. Seal with neutral gum and observe under a microscope.

[0061] 2.6 Data Processing GrapaPad Prism software was used for one-way analysis of variance (ANOVA) to compare the weight, food intake, and animal organ index of mice among the groups. The data processing results were expressed as Mean ± SEM. p When the value was <0.05, it was considered to be statistically significant.

[0062] 3. Experimental results 3.1 Effect of Codonopsis pilosula aqueous extract on body weight in mice with high-fat diet-induced obesity The condition of the mice was observed daily. During the administration period, the diet and activity of the mice given the Codonopsis pilosula water extract were normal.

[0063] The weight of mice was recorded every week and a weight change curve was drawn. Figure 2 and Figure 3 As shown, compared with the HFD group, the DS group could delay the increase of body weight in mice.

[0064] Effect of Codonopsis pilosula aqueous extract on food intake in mice with high-fat diet-induced obesity The food intake of mice was recorded daily and a graph of the changes in food intake was drawn. Figure 4 and Figure 5 As shown, the water extract of Codonopsis pilosula can reduce the appetite and food intake of mice.

[0065] 3.3 Effect of Codonopsis pilosula extract on liver histopathology in mice with high-fat diet-induced obesity H&E staining results ( Figure 6 ) showed that the number of lipid droplets increased in the high-fat group, while the number of lipid droplets in the Codonopsis pilosula water extract group was significantly reduced after administration.

[0066] 3.4 Effect of Codonopsis pilosula aqueous extract on leptin levels in mice with high-fat diet-induced obesity Elisa results ( Figure 7 ) showed that the leptin level in the serum of mice in the high-fat group was increased, and the leptin level also showed an upward trend after the administration of Codonopsis pilosula water extract.

[0067] RT-qPCR results ( Figure 8 ) showed that the leptin level in the hypothalamus of mice in the high-fat group was decreased, and the leptin level was upregulated after administration of Codonopsis pilosula water extract.

[0068] 3.5 Effect of Codonopsis pilosula aqueous extract on the level of hypothalamic feeding neurons in mice with high-fat diet-induced obesity RT-qPCR results ( Fig. 9 and Fig.10 ) showed that the mRNA expression levels of Agrp and NPY in the hypothalamic feeding neurons of mice in the high-fat group were increased, while the mRNA expression levels of Agrp and NPY in the Codonopsis pilosula water extract group were significantly decreased after administration.

[0069] Example 2 This embodiment provides a functional food that helps control body fat, wherein the active ingredient is water extract of Codonopsis pilosula, and the auxiliary materials are auxiliary materials acceptable to functional foods.

[0070] For example, the dosage form of the functional food can be selected as a granule, which is made of the following raw materials by mass: 80 parts of Codonopsis pilosula water extract, 20 parts of microcrystalline cellulose, 5 parts of low-substituted hydroxypropyl cellulose, 0.5 parts of flavor, 1 part of aspartame, and 15 parts of 5% PVP aqueous solution. The preparation method is: mix the Codonopsis pilosula water extract with microcrystalline cellulose, add 5% PVP aqueous solution to wet granulate with a 20-mesh sieve, dry the granules at 40-55°C until the moisture content is less than 2%, and then sieve through an 18-mesh sieve, add flavor and aspartame, mix evenly, and package them.

[0071] Example 3 This embodiment provides a medicine for treating food-induced obesity or alleviating the symptoms of food-induced obesity, wherein the active ingredient is the water extract of Codonopsis pilosula, and the excipient is a pharmaceutically acceptable excipient for oral preparations.

[0072] For example, the drug is a capsule, which is made of the following raw materials by mass: 80 parts of Codonopsis pilosula water extract, 30 parts of pregelatinized starch, 15 parts of cross-linked polyvinyl pyrrolidone, and 15 parts of hydroxypropyl methylcellulose. The Codonopsis pilosula water extract, pregelatinized starch, hydroxypropyl methylcellulose and 10 parts of cross-linked polyvinyl pyrrolidone are mixed, granulated in one step with water, the obtained granules are dried to a moisture content of <2%, and then granulated, the remaining cross-linked polyvinyl pyrrolidone is added, mixed evenly, and capsules are filled on an automatic capsule filling machine, and then packaged to obtain the drug.

[0073] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of Codonopsis pilosula water extract in the preparation of products for alleviating or treating food-induced obesity.

2. The use according to claim 1, characterized in that: The preparation method of the water extract of Codonopsis pilosula is as follows: crush Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

3. The use according to claim 2, characterized in that: Crush Codonopsis pilosula to a particle size of <2 mm; and / or The heating temperature is 95-105°C; and / or Soak the crushed Codonopsis pilosula in water for 1 hour before heating for extraction; and / or Extract twice with water and combine the extracts; in each extraction, the mass ratio of ginseng to water is 1:8-12; and / or The drying temperature is 45-55°C.

4. A functional food that helps control body fat, characterized in that: Its active ingredients include Codonopsis pilosula water extract.

5. The functional food according to claim 4, characterized in that The preparation method of the water extract of Codonopsis pilosula is as follows: crush Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

6. The functional food according to claim 5, characterized in that Crush Codonopsis pilosula to a particle size of <2 mm; and / or The heating temperature is 95-105°C; and / or Soak the crushed Codonopsis pilosula in water for 1 hour before heating for extraction; and / or Extract twice with water and combine the extracts; in each extraction, the mass ratio of ginseng to water is 1:8-12; and / or The drying temperature is 45-55°C.

7. A medicine for treating food-induced obesity or alleviating the symptoms of food-induced obesity, characterized in that: Its active ingredients include Codonopsis pilosula water extract.

8. The medicine according to claim 7, characterized in that The drug is a drug for reducing leptin resistance; and / or The medicine is an oral preparation.

9. The medicine according to claim 7 or 8, characterized in that: The preparation method of the water extract of Codonopsis pilosula is as follows: crush Codonopsis pilosula, heat it with water for extraction, and concentrate and dry the obtained extract.

10. The drug according to claim 9, characterized in that Crush Codonopsis pilosula to a particle size of <2 mm; and / or The heating temperature is 95-105°C; and / or Soak the crushed Codonopsis pilosula in water for 1 hour before heating for extraction; and / or Extract twice with water and combine the extracts; in each extraction, the mass ratio of ginseng to water is 1:8-12; and / or The drying temperature is 45-55°C.

Images