Composition with effects of reducing fat and losing weight as well as preparation method and application of composition

Abstract

The invention discloses a composition with lipid-lowering and weight-losing effects as well as a preparation method and application thereof, and belongs to the technical field of biomedical treatment. The invention aims to provide a composition which is more suitable for people. The invention provides a composition with lipid-lowering and weight-losing effects. The pharmaceutical composition is prepared from the following components: 1-50 parts of radix astragali, 1-50 parts of pericarpium citri reticulatae, 1-50 parts of fructus crataegi and 1-50 parts of medicated leaven. The invention provides a lipid-lowering pharmaceutical composition with higher practicability.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and specifically relates to a composition with lipid-lowering and weight-loss effects, and a preparation method and application thereof. Background Art

[0002] Obesity has become the root cause of various diseases in today's society, so the prevention and treatment of obesity has also become a major issue in population health. The drugs or health products currently on the market for the treatment of obesity have certain defects, the specific defects are as follows:

[0003] ① Traditional lipid-lowering drugs and health products have limited effects

[0004] Most of the lipid-lowering products available on the market rely on a single chemical drug ingredient or an ordinary herbal formula. Although they have certain efficacy, they are not effective enough in regulating the balance of lipid metabolism in the body and maintaining healthy blood lipids in the long term. Traditional Chinese medicine formulas have failed to fully integrate modern pharmacological research, resulting in a lack of scientific data support for efficacy evaluation.

[0005] ②Narrow scope of applicable population

[0006] Existing lipid-lowering products are not targeted enough for specific groups (such as patients with hyperlipidemia or obese people). For example, patients with diabetes or metabolic syndrome are restricted in choosing products containing sugar or irritating ingredients. In addition, there is a problem of limited age coverage and failure to effectively take into account the different needs of young people, the elderly and sub-healthy people.

[0007] ③The dosage form is single and not portable enough

[0008] Currently, most lipid-lowering products on the market are in the form of traditional medicinal soups or capsules, with a single usage scenario and a lack of portable designs suitable for modern fast-paced life, such as effervescent tablets, oral liquids or ready-to-drink beverages.

[0009] ④ Lack of significant and scientific data support

[0010] The experimental data on functional verification of existing products (such as the regulatory effects on lipid metabolism) are often insufficient or lack scientific basis, making it difficult to form authoritative endorsement, which affects consumer trust and market promotion.

[0011] ⑤Lack of environmental protection and naturalness

[0012] Some existing technologies are not green and environmentally friendly in terms of raw material selection and production processes. There may be problems with chemical additives or improper disposal of production waste, and they cannot meet the demand of modern consumers for "natural and healthy" products.

[0013] ⑥Weak market targeting

[0014] Existing technologies are difficult to meet the specific needs of modern urban people (such as obesity and dyslipidemia caused by long periods of sitting in the office and lack of exercise), and lack precise design for segmented markets.

[0015] Therefore, a more practical lipid-lowering drug combination is urgently needed. Summary of the invention

[0016] The purpose of the present invention is to provide a composition suitable for a wider range of people.

[0017] The invention provides a composition with lipid-lowering and weight-loss effects. The drug combination consists of the following ingredients: 1-50 parts of astragalus, 1-50 parts of tangerine peel, 1-50 parts of hawthorn and 1-50 parts of Shenqu.

[0018] It is further defined that the drug combination is composed of the following ingredients: 10 parts of Astragalus, 10 parts of Tangerine Peel, 10 parts of Crataegus and 10 parts of Shenqu.

[0019] It is further defined that the drug combination is composed of the following ingredients in a ratio of: Astragalus: Tangerine Peel: Hawthorn: Shenqu = 1:1:1:1.

[0020] The invention provides a method for preparing the above-mentioned composition, and the specific steps of the preparation method are as follows: astragalus, tangerine peel, hawthorn, and Shenqu are weighed in a ratio of 1:1:1:1, the above-mentioned medicinal materials are added with 10 times the volume of water and soaked for 1 hour, boiled with high heat, then decocted with low heat for 1 hour, filtered while hot, and the medicinal residues are added with 8 times the volume of water and continued to be decocted for 1 hour, and the medicinal liquids obtained by two filtrations are combined and concentrated to 1.47 g / mL.

[0021] The present invention provides an application of the above-mentioned composition in preparing drugs for reducing lipids, blood sugars and body weight.

[0022] The present invention provides an application of the above-mentioned composition in preparing a weight-loss drug.

[0023] The present invention provides an application of the above-mentioned composition in preparing a medicine for reducing the TG content in blood.

[0024] The present invention provides an application of the above-mentioned composition in preparing a medicine for reducing the LDH content in cells.

[0025] The present invention provides an application of the above-mentioned composition in preparing a medicine for increasing the content of GSH and SOD in cells.

[0026] It is further defined that the dosage form of the composition is any one of tablets, capsules, granules, powders, and liquid preparations.

[0027] Beneficial effects: Obesity and hyperlipidemia mostly belong to the category of "cream" and "phlegm" in traditional Chinese medicine. Irregular diet and overeating of fat and sweet foods lead to spleen deficiency, spleen dysfunction, and weakened function of transporting water and grain. Sedentary and lack of exercise and other lifestyles further aggravate the stagnation of qi, resulting in the inability of water and fluid to flow, the accumulation of body fluids, and the accumulation of phlegm and fat in the body. In severe cases, blood stasis blocks the blood vessels. Treatment should focus on strengthening the spleen and regulating qi, removing dampness and removing turbidity. In the prescription, Astragalus is the main drug, which can replenish the middle qi, invigorate qi and raise yang, and restore the healthy operation of the spleen; the present invention is that tangerine peel is the minister drug, which can regulate qi and strengthen the spleen, dry dampness and resolve phlegm, and is compatible with Astragalus, combining tonification and tonification, and tonifying without stagnation; Hawthorn can help digestion and resolve accumulation, promote qi and disperse blood stasis, and Shenqu can help digestion and harmonize the stomach, which are adjuvants. The four drugs are used together to play the effects of strengthening the spleen and regulating qi, removing dampness and removing turbidity.

[0028] There is no effective preventive medicine for high-risk groups of hyperlipidemia. Traditional Chinese medicine can reduce obesity and lower blood lipids, but it is inconvenient to boil traditional Chinese medicine decoctions and tastes bitter. The invention has a simple formula, a sweet taste, and adopts a tea-substitute dosage form, which is convenient for long-term use. The preparation method is as follows: take each raw medicinal material, grind it into small pieces with a special mill for traditional Chinese medicine, use disposable non-woven fabric to make tea bags, each bag contains about 40g of traditional Chinese medicine powder, brew with 300-500mL boiling water, drink it warm instead of tea, and take 1 bag each time before meals. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The results of the effect of the traditional Chinese medicine formula on the body weight of NAFLD rats; A is the result before administration; B is the result after administration;

[0030] Figure 2 The results of the effect of the traditional Chinese medicine formula on serum TG and TC in NAFLD rats are shown in Figure 1. A is the results of the TC content in rat serum; B is the results of the TG content in rat serum;

[0031] Figure 3 The results of the effect of traditional Chinese medicine formula on liver tissue pathology in NAFLD rats (200×);

[0032] Figure 4 This is the result diagram of the effect of traditional Chinese medicine formula on liver tissue of NAFLD rats;

[0033] Figure 5 This is the Oil Red O staining diagram of the FFA-induced HepG2 cells by the Chinese medicine formula; A is the staining result diagram; B is the absorbance result diagram of the Oil Red O staining of the Chinese medicine formula;

[0034] Figure 6 The figure is a result of the determination of the content of triglyceride (TG) and cholesterol (TC) in HepG2 cells induced by FFA by the traditional Chinese medicine formula; A is the result of the TC content in the 12% serum system; B is the result of the TG content in the 12% serum system;

[0035] Figure 7The results of the effect of the traditional Chinese medicine formula on the oxidative stress level in HepG2 cells induced by FFA are shown in Figure 1; A is the results of the SOD content in the 12% serum system; B is the results of the GSH content in the 12% serum system; C is the results of the LDH content in the 12% serum system;

[0036] Figure 8 A is a graph showing the effect of the traditional Chinese medicine formula on the level of liver damage in HepG2 cells induced by FFA; A is a graph showing the ALT content in a 12% serum system; and B is a graph showing the AST content in a 12% serum system. DETAILED DESCRIPTION

[0037] Example 1. A composition having lipid-lowering and weight-loss effects

[0038] 1. Recipe: Astragalus: 10g, Tangerine Peel: 10g, Hawthorn: 10g, Shenqu: 10g.

[0039] 2. Preparation method: Weigh astragalus, tangerine peel, hawthorn, and Shenqu in a ratio of 1:1:1:1. Add 10 times the volume of water to soak the above medicinal materials for 1 hour, boil over high heat, then simmer for 1 hour over low heat, filter while hot, add 8 times the volume of water to the residue and continue to boil for 1 hour, combine the medicinal liquids obtained by two filtrations, and concentrate to 1.47 g / mL.

[0040] Example 2. Application of the composition

[0041] 1. Experimental methods: The NAFLD rat model (SD male) was established by feeding with a high-fat diet for 45 days. The rats were divided into 5 groups, including a blank group (normal SD rats), a model group (NAFLD rat model), a positive drug Yishanfu group (142.5 mg / kg), a low-dose group (3.597 g / kg), and a high-dose group (7.194 g / kg). The drugs were given once a day for 30 consecutive days. The HE staining of the liver and the changes in liver morphology after the intervention of the Chinese medicine formula were observed. The levels of triglyceride (TG) and cholesterol (TC) in serum were detected by an enzyme marker to evaluate the improvement effect of the Chinese medicine formula on the NAFLD rat model.

[0042] High-fat feed formula for rats: 2.5% cholesterol, 0.5% sodium cholate, 5% lard, 10% egg yolk powder, and 82% basic feed.

[0043] 2. Experimental results: Effects of Chinese herbal medicine formula on body weight of NAFLD rats

[0044] Experimental method: The NAFLD rat model (SD male - initial body weight 180g) was established by feeding with high-fat diet for 45 days. The rats were divided into 5 groups, blank group (normal SD rats), model group (NAFLD rat model), positive drug Yishanfu group (142.5mg / kg Yishanfu), low-dose group (Chinese medicine formula 3.597g / kg), and high-dose group (Chinese medicine formula 7.194g / kg). The drugs were administered once a day for 30 consecutive days.

[0045] Results: Compared with the blank group, there was no significant difference in the weight gain of the rats in the model group. The reason for this was that the rats resisted eating high-fat feed in the later stage of high-fat feeding, resulting in no increase in weight. However, compared with the model group, the weight of the rats in the Chinese medicine formula group decreased. Figure 1 .

[0046] 3. Effect of Chinese herbal medicine formula on lipid accumulation in NAFLD rats

[0047] Compared with the blank group, the serum TC and TG levels of the model group rats were significantly increased; compared with the model group, the Chinese medicine formula could significantly reduce the serum TC level of NAFLD rats, and the serum TG level of the low-dose and high-dose groups rats was significantly reduced. Figure 2 .

[0048] 4. Effects of Chinese medicine formula on liver tissue pathology in NAFLD rats

[0049] Experimental method: After the administration, rats were anesthetized and dissected with 1% sodium pentobarbital solution. The liver lobules of 6 rats were randomly selected from each group and immersed in 4% paraformaldehyde for fixation for more than 48 hours. The samples were then placed in xylene for 10 minutes, dried, placed in xylene again for 10 minutes, dried, and placed in anhydrous ethanol for 5 minutes. After drying, they were placed in 95%, 90%, 80%, and 70% alcohol in sequence, each time immersed for 5 minutes. After the operation was completed, they were washed with distilled water; the washed samples were placed in hematoxylin for 8 minutes and rinsed with tap water. Soak in 1% hydrochloric acid for 30 seconds, rinse with tap water, and after rinsing, use 0.6% ammonia water for blueing treatment, and then treat with tap water; put the treated slices into eosin staining solution for 3 minutes and rinse with water; soak the treated slices in 95% alcohol for 5 minutes, soak them in 95% alcohol for 5 minutes after drying, and then soak them in anhydrous ethanol for 5 minutes, repeat twice; soak them in xylene for 5 minutes after drying, repeat twice, and seal them with neutral gum after drying; use a microscope to examine the sample slices and collect data for analysis.

[0050] According to the results of HE staining, the liver tissue structure of the blank group was clear, while the liver cells in the model group were enlarged and round fat vacuoles of different sizes were visible in the cytoplasm; and the liver tissue damage in the Chinese medicine formula group was alleviated compared with the model group. Figure 3 .

[0051] According to the results of macroscopic observation of liver appearance, there were no abnormal changes in the liver of the blank group. The liver of the model group was significantly enlarged, with a tight capsule, rounded edges, milky white color, and greasy cut surface. The damage in the positive drug group and the Chinese medicine formula was improved after administration. Figure 4 .

[0052] In vitro experiments

[0053] Experimental methods: The lipid accumulation cell model was established by using FFA-induced human hepatoma cells (HepG2) to investigate the effect of Chinese medicine formula on NAFLD from the aspects of lipid accumulation, oxidative stress and liver cell damage. The efficacy evaluation indexes included intracellular triglyceride (TG), total cholesterol (TC), superoxide dismutase (SOD), glutathione (GSH), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels, and the intracellular lipid accumulation level was determined by combining the intracellular lipid droplet formation with Oil Red O staining experiment.

[0054] 5. Effects of Chinese herbal medicine formula on lipid accumulation in HepG2 cells induced by FFA

[0055] Experimental methods:

[0056] (1) Cell Oil Red O staining experiment

[0057] HepG2 cells were cultured at 40×10 4The cells were inoculated into 6-well plates at a density of 100 cells / well and cultured in a cell culture incubator at 37°C and 5% CO2. The experimental cells were divided into 5 groups, namely, blank group, model group (1mM FFA), low-dose group (1mM FFA + 3% drug-containing serum), medium-dose group (1mM FFA + 6% drug-containing serum), and high-dose group (1mM FFA + 12% drug-containing serum). According to the above grouping, the corresponding concentrations of drugs and modeling agents were added to treat the cells for 24 hours. Each well was repeated 6 times. Except for the different induction conditions and drug interventions, the other conditions were parallel. After the drug treatment, Oil Red O staining was performed. The specific operation is as follows: discard the culture medium, wash with PBS 3 times, and fix with 4% paraformaldehyde for 30 minutes. Wash twice with PBS buffer. Under light-proof and sealed conditions, add Oil Red O staining working solution and incubate for 20 minutes, rinse with 60% isopropanol for 30 seconds, and differentiate to interstitial washing. Rinse the cells with PBS 3 times, 1 minute each time. The cells were stained with hematoxylin solution for 2 min, washed with PBS 3 times, 1 min each time, sealed with glycerol gelatin, and finally placed under an optical microscope to observe the red lipid droplets in the cells and collect pictures.

[0058] (2) Determination of intracellular TG and TC indicators

[0059] HepG2 cells were cultured at 40×10 4 The cells were inoculated into 6-well plates at a density of 100 cells / well and cultured in a cell culture incubator at 37°C and 5% CO2. The experimental cells were divided into 5 groups, namely, blank group, model group (1mM FFA), low-dose group (1mM FFA + 3% drug-containing serum), medium-dose group (1mM FFA + 6% drug-containing serum), and high-dose group (1mM FFA + 12% drug-containing serum). According to the above grouping, the corresponding concentrations of drugs and modeling agents were added to treat the cells for 24 hours. Each well was repeated 6 times. Except for the different induction conditions and drug interventions, the other conditions were parallel. After the cell treatment was completed, the cells were ultrasonically lysed, centrifuged at 4°C and 12000rpm / min for 10 minutes, and the cell supernatant of each group was collected and stored for later use. The cell lysate samples prepared in each group were taken, and the protein quantitative BCA method was used to determine the intracellular protein content of each group. According to the instructions of the corresponding detection kit, the CHOD-PAP enzymatic method was used to detect the levels of triglycerides (TG) and total cholesterol (TC) in each group of cells.

[0060] Oil Red O can bind to triglycerides in cells and dye triglycerides red and orange, and hematoxylin can dye cell nuclei blue-purple. Using Oil Red O to stain cells can determine the fat content in cells. In order to study whether the Chinese medicine formula has an improvement effect on the lipid accumulation of HepG2 cells induced by FFA, we first used Oil Red O to observe the effect of the Chinese medicine formula on the formation of red lipid droplets in HepG2 cells induced by FFA, and detected the effect of the Chinese medicine formula on the TG and TC levels in HepG2 cells induced by FFA.

[0061] Oil red O staining results showed that ( Figure 5 ), and there were almost no red lipid droplets in the blank group. After 24 hours of FFA induction, a large number of red lipid droplets appeared in the FFA group cells, indicating that FFA can cause lipid accumulation in HepG2 cells. After intervention with the Chinese medicine formula, the red lipid droplets in the cells became lighter in color than those in the FFA group, and the diffusion range of lipid droplets was reduced, indicating that the accumulation of cellular lipids was improved after drug treatment, and it was dose-dependent.

[0062] At the same time, the improvement effect of the traditional Chinese medicine formula on the lipid accumulation of HepG2 cells induced by FFA was evaluated by detecting the TG and TC contents in each group of cells. The results showed that compared with the blank group, the TG and TC contents of HepG2 cells induced by FFA administration increased, and the differences were statistically significant (P<0.05), indicating that the FFA-induced cell lipid accumulation model was successfully established. After intervention with different doses of traditional Chinese medicine formula, high doses can significantly reduce TG content, and high doses can significantly reduce TC content ( Figure 6 ).

[0063] Combined with the results of cell oil red O staining, TG and TC content determination, the effect of traditional Chinese medicine formula on lipid accumulation of HepG2 cells induced by FFA was comprehensively evaluated. It showed that traditional Chinese medicine formula could reduce the TG and TC content of HepG2 cells induced by FFA, reduce the formation of intracellular lipid droplets, and thus achieve the effect of reducing lipid accumulation, and the improvement effect of traditional Chinese medicine formula on lipid accumulation was the most significant.

[0064] 6. Effect of Chinese herbal medicine formula on FFA-induced oxidative stress level in HepG2 cells

[0065] Experimental method: HepG2 cells were cultured at 40×10 4The cells were inoculated into 6-well plates at a density of 100 cells / well and cultured in a cell culture incubator at 37°C and 5% CO2. The experimental cells were divided into 5 groups, namely, blank group, model group (1mM FFA), low-dose group (1mM FFA + 3% drug-containing serum), medium-dose group (1mM FFA + 6% drug-containing serum), and high-dose group (1mM FFA + 12% drug-containing serum). According to the above grouping, the corresponding concentrations of drugs and modeling agents were added to treat the cells for 24 hours. Each well was repeated 6 times. Except for the different induction conditions and drug interventions, the other conditions were parallel. After the cell treatment was completed, the cells were ultrasonically lysed and centrifuged at 4°C and 12000rpm / min for 10 minutes. The cell supernatant of each group was collected and stored for later use. The cell lysate samples prepared in each group were taken, and the protein content in each group of cells was determined by protein quantitative BCA method. According to the instructions of the corresponding detection kits, the levels of SOD, GSH and LDH in the cells of each group were detected (using the xanthine oxidase method, dithiodinitrobenzoic acid method and 2,4-dinitrophenylhydrazine method, respectively).

[0066] Oxidative stress plays an important role in the occurrence and development of NAFLD. However, when free fatty acids and their metabolites in the liver exceed the metabolic load of cells, they will induce cells to produce a large amount of reactive oxygen species (ROS). Intracellular ROS can be quickly cleared by a variety of antioxidant enzymes to prevent redox damage to biological molecules and cells. ROS can be mediated by antioxidant pathways such as superoxide dismutase (SOD) and glutathione (GSH), among which GSH and SOD are antioxidant reducing agents, and LDH reflects the degree of damage to the body. In order to study whether the Chinese medicine formula can improve the antioxidant capacity of HepG2 cells induced by FFA, we detected the content of antioxidant enzymes SOD, GSH and LDH in each group of cells.

[0067] The results of intracellular GSH and SOD detection showed that the intracellular antioxidant indexes (GSH, SOD) in the FFA group were lower than those in the blank group. After intervention with the Chinese medicine formula, the intracellular GSH and SOD contents were significantly increased (P < 0.01). In addition, the Chinese medicine formula dose-dependently reduced the intracellular LDH level (P < 0.01) ( Figure 7 ).

[0068] Therefore, this experiment comprehensively evaluated the effect of Chinese medicine formula on the oxidative stress level of HepG cells induced by FFA by measuring the content of oxidative stress indicators (GSH, SOD and LDH) in FFA-induced cells. The above results show that Chinese medicine formula can accelerate the removal of intracellular ROS and reduce intracellular oxidative stress by increasing the levels of intracellular antioxidant enzymes GSH and SOD in the FFA-induced cell model.

[0069] 7. Effects of Chinese herbal medicine formula on FFA-induced liver injury in HepG2 cells

[0070] Experimental method: HepG2 cells were cultured at 40×10 4 The cells were inoculated into 6-well plates at a density of 100 cells / well and cultured in a cell culture incubator at 37°C and 5% CO2. The experimental cells were divided into 5 groups, namely, blank group, model group (1mM FFA), low-dose group (1mM FFA + 3% drug-containing serum), medium-dose group (1mM FFA + 6% drug-containing serum), and high-dose group (1mM FFA + 12% drug-containing serum). According to the above grouping, the corresponding concentrations of drugs and modeling agents were added to treat the cells for 24 hours. Each well was repeated 6 times. Except for the different induction conditions and drug interventions, the other conditions were parallel. After the cell treatment was completed, the cells were ultrasonically lysed, centrifuged at 4°C and 12000rpm / min for 10 minutes, and the cell supernatant of each group was collected and stored for later use. Take the prepared cell lysate samples of each group, and use the protein quantitative BCA method to determine the intracellular protein content of each group. According to the instructions of the corresponding detection kit, the AST level in each group of cells was detected by immunosuppression method, and the ALT level was detected by pyruvate oxidase method.

[0071] In order to study whether the Chinese medicine formula can improve the damage of HepG2 cells treated with FFA, we detected the intracellular ALT and AST levels under the FFA-induced model. Figure 8 ), and the high-dose group had a significant difference in the regulation of ALT (P<0.05). This proves that the Chinese medicine formula can play a good protective role on liver cell function and reverse the liver cell function damage caused by the lipid accumulation cell model induced by mixed fatty acids.

Claims

1. A composition having lipid-lowering and weight-loss effects, characterized in that: The drug combination is composed of the following ingredients: 1-50 parts of astragalus, 1-50 parts of tangerine peel, 1-50 parts of hawthorn and 1-50 parts of Shenqu.

2. The composition according to claim 1, characterized in that The drug combination is composed of the following ingredients: 10 parts of astragalus, 10 parts of tangerine peel, 10 parts of hawthorn and 10 parts of Shenqu.

3. The composition according to claim 1, characterized in that The drug combination is composed of the following ingredients in a ratio of astragalus: tangerine peel: hawthorn: Shenqu = 1:1:1:

1.

4. A method for preparing the composition according to any one of claims 1 to 3, characterized in that: The specific steps of the preparation method are as follows: astragalus, tangerine peel, hawthorn, and Shenqu are weighed in a ratio of 1:1:1:1, the above medicinal materials are added with 10 times the volume of water and soaked for 1 hour, boiled with high heat, then decocted with low heat for 1 hour, filtered while hot, and the residues are added with 8 times the volume of water and continued to be decocted for 1 hour, and the medicinal liquids obtained by two filtrations are combined and concentrated to 1.47 g / mL.

5. Use of the composition according to any one of claims 1 to 3 in the preparation of drugs for reducing lipids, blood sugars and body weight.

6. Use of the composition according to any one of claims 1 to 3 in the preparation of weight loss drugs.

7. Use of the composition according to any one of claims 1 to 3 in the preparation of a drug for reducing TG content in blood.

8. Use of the composition according to any one of claims 1 to 3 in the preparation of a drug for reducing LDH content in cells.

9. Use of the composition according to any one of claims 1 to 3 in the preparation of a drug for increasing the content of GSH and SOD in cells.

10. The use according to any one of claims 5 to 9, characterized in that: The dosage form of the composition is any one of tablets, capsules, granules, powders and liquid preparations.

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