Application of mallotus oblongifolius ethanol extract in reducing fat and losing weight

Abstract

The invention belongs to the field of food nutritional supplements and medicines, and particularly relates to application of a mallotus oblongifolius ethanol extract in the aspects of lowering lipid and losing weight. The invention aims to provide the mallotus oblongifolius ethanol extract which can effectively reduce weight gain of mice induced by high-fat and high-cholesterol diet, reduce fat content of the mice, reduce the surface area of fat cells and increase heat production of the mice, so that the mallotus oblongifolius ethanol extract has a remarkable treatment effect on weight loss. The traditional Chinese medicine composition overcomes the defects of scarce selection of weight-reducing medicines and high cost and adverse drug reaction caused by the existing medicines clinically at present, is safe and effective, provides a new choice for preventing / treating obesity, and is simple in preparation method and low in cost.

Description

Technical Field

[0001] The invention belongs to the field of food nutritional supplements and medicines, and more specifically, relates to an application of a partridge tea ethanol extract in reducing fat and weight. Background Art

[0002] Obesity, usually defined as excessive accumulation of adipose tissue, has become a global health problem affecting all ages and genders. Further development of obesity will lead to a variety of metabolic diseases. In order to control obesity, people usually use lifestyle interventions, and choose to use drug treatment when the effect is not good. The types of chemical synthetic lipid-lowering and weight-loss drugs currently used in clinical practice are limited, mainly including orlistat, liraglutide and other major categories. There are common side effects such as gastrointestinal reactions, liver damage, cardiovascular risks, and long-term use is prone to drug resistance. Although the safety of traditional Chinese medicine weight-loss preparations has been improved, there are problems such as complex ingredients, unclear effective material basis, and vague mechanism of action, which make it difficult to meet the needs of modern precision medicine. With the global incidence of obesity and metabolic syndrome rising year by year, the development of new natural-source weight-loss drugs with significant efficacy, clear targets and good safety has become an urgent need in the medical field.

[0003] As a characteristic medicinal material system of Lingnan, "Southern medicine" has formed the characteristics of highly active secondary metabolites enrichment due to the unique tropical island ecological environment. Partridge tea is a perennial evergreen shrub or small tree, distributed in Sanya, Lingshui, Wanning, Wenchang, Ding'an, Dongfang and Baoting in Hainan Province. Its leaves are round and sweet, and it has a unique flavor when brewed and drunk. It has been praised by literati and poets of all dynasties as "Ganoderma lucidum grass", becoming a major cultural feature of Hainan local tea. As a characteristic medicinal and edible plant of Hainan, it was recorded as clearing away heat and dampness, digesting food and relieving stagnation as early as in the "Supplement to Compendium of Materia Medica". Its safety has been verified by hundreds of years of folk drinking practice. Modern research shows that it is rich in polyphenols, flavonoids, and triterpenoids, and has the potential to regulate lipid metabolism and inhibit the differentiation of adipocytes. Compared with conventional drug development paths, developing weight loss products using partridge tea as raw material has three core advantages: first, its medicinal and edible properties allow it to avoid the risk of toxic side effects of synthetic drugs; second, its unique southern medicinal resources give it novel ingredients, which is conducive to the discovery of new targets; third, the ethanol extraction process can directionally enrich fat-soluble active ingredients, laying the foundation for subsequent preparation standardization.

[0004] Since existing weight loss methods all have certain difficulties and risks, a preparation method of quail tea ethanol extract and its application in lipid-lowering and weight loss are proposed, which provides a safe and effective option for the treatment of obesity and also provides a theoretical basis for the development of quail tea as a special resource with high potential application value in Hainan. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the existing high-safety and significant-effect weight-loss drugs in short supply, and to provide a use of a partridge tea ethanol extract with high safety and good lipid-lowering effect in the preparation of weight-loss drugs.

[0006] Another object of the present invention is to provide a method for preparing the ethanol extract of Partridge Tea.

[0007] The above-mentioned purpose of the present invention is achieved through the following technical solutions: S1, drying the partridge tea material and crushing it into powder; S2, soaking with 12% ethanol, and filtering the extract after soaking for three days, and the above operation is performed three times, and the obtained filtrate is the ethanol extract of partridge tea; S3. Concentrate the filtrate under vacuum at 45°C to obtain an extract.

[0008] Furthermore, an application of a partridge tea ethanol extract is characterized in that the extract is used to treat or prevent obesity and related symptoms caused by a high-fat and high-cholesterol diet.

[0009] Furthermore, the pharmaceutical composition of the partridge tea extract is characterized in that the composition contains the partridge tea ethanol extract as an active ingredient.

[0010] In addition, the present invention also provides a specific application method of the ethanol extract of Partridge Tea, which specifically comprises the following steps: S1. Purchase 32 8-week-old male C57BL / 6J mice weighing 18-20 g at the specific pathogen-free (SPF) level; S2. Mice were raised under constant temperature and humidity, and divided into 4 groups (8 mice in each group) according to body weight after one week: normal control group, high-fat and high-cholesterol diet model group, partridge tea ethanol extract intervention group, and positive drug rosiglitazone intervention group; S3. Except for the normal control group, the other groups were given a high-fat diet and given drugs by oral gavage or intraperitoneal injection at 10 a.m. every day. The body weight of the mice was observed and recorded for 60 consecutive days. S4, after step S3, anesthetize and collect blood, kill the mice by cervical dislocation, and dissect them, collect and record the weight of the liver and adipose tissue of the mice; S5, weighing a certain mass of the mouse subcutaneous fat tissue, abdominal white fat tissue, and brown fat tissue collected in step S4, making paraffin sections, staining with hematoxylin-eosin (HE), and observing pathological sections; S6, performing immunohistochemical staining of UCP1 on the paraffin sections of brown adipose tissue prepared in S5, and performing section observation; Furthermore, in step S2, the extract is administered at a dosage of 500 mg / kg, and the positive drug group is administered at a dosage of 10 mg / kg.

[0011] Furthermore, in step S3, the high-fat and high-cholesterol feed formula contains 60% fat and 1.2% cholesterol.

[0012] The present invention has the following advantages: In the first aspect of the present invention, the ethanol extract of Partridge Tea can significantly slow down the weight gain of mice, and the effect is consistent with that of the positive drug group.

[0013] In a second aspect of the present invention, the ethanol extract of Partridge Tea can significantly reduce the weight of subcutaneous adipose tissue, abdominal white adipose tissue and brown adipose tissue in mice.

[0014] In a third aspect of the present invention, the ethanol extract of Partridge Tea can reduce the size of lipid droplets in adipose tissue of mice.

[0015] In a fourth aspect of the present invention, the ethanol extract of Partridge Tea can promote the expression of UCP1 in brown adipose tissue of mice and increase the heat production of mice.

[0016] The fifth aspect of the present invention provides a medicinal and edible extract which is of natural origin, highly safe and has few side effects.

[0017] A sixth aspect of the present invention provides a use of an ethanol extract of Partridge Tea in a preparation for preventing obesity-related diseases caused by a high-fat, high-cholesterol diet.

[0018] A seventh aspect of the present invention is a preparation for preventing diseases related to hyperlipidemia, comprising an ethanol extract of Partridge Tea.

[0019] In the eighth aspect of the present invention, partridge tea is a new resource food, and its extract has little toxicity and side effects, is low in price, is easy to take, and can be used for a long time. It is expected to become a safer and more reliable drug for improving obesity that can be taken for a long time.

[0020] The present invention has the following beneficial effects: The present invention provides an application of ethanol extract of quail tea in the preparation of lipid-lowering and weight-loss drugs. Animal experiments on obese mice induced by a high-fat and high-cholesterol diet verify that the extract has a good weight-loss effect. Compared with the clinical drug rosiglitazone, the ethanol extract of quail tea not only reduces the weight of mice but also alleviates liver damage. It has great potential for the development of weight-loss and liver-protecting drugs, providing more options for patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a graph showing the weight changes of mice in the present invention. Figure 2This is a statistical graph of the weight of subcutaneous fat tissue in mice after 8 weeks of drug administration according to the present invention. Figure 3 This is a statistical graph of the weight of white adipose tissue in the abdomen of mice after 8 weeks of drug administration according to the present invention. Figure 4 This is a statistical graph of the weight of brown adipose tissue in mice after 8 weeks of drug administration according to the present invention. Figure 5 This is an immunohistochemical staining image of UCP1 in brown adipose tissue of mice after 8 weeks of drug administration according to the present invention. Figure 6 This is a real shot of the mouse liver after 8 weeks of drug administration and a HE staining of adipose tissue under an optical microscope. DETAILED DESCRIPTION

[0022] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0023] Example 1 Preparation of Partridge Tea Ethanol Extract The preparation of the ethanol extract of partridge tea comprises the following steps: S1. Weigh 2 kg of dried partridge tea.

[0024] S2. Grind the partridge tea in step S1 into fine powder using a grinder.

[0025] S3. Place the partridge tea powder obtained in step S2 into a 10 L container and add a sufficient amount of 12% ethanol to ensure that the powder is completely covered by ethanol.

[0026] S4. Seal the container and soak it in a constant temperature environment for three days.

[0027] S5. After soaking for three days, filter the solution using a 2 L suction filtration device to remove undissolved solid residues.

[0028] S6. To improve the extraction efficiency, the filtration process in step S5 was repeated three times, and the filtrate was replaced with new 12% ethanol each time to ensure that as many effective ingredients as possible were extracted.

[0029] S7. Combine the filtrates obtained from the three filtrations.

[0030] S8. Use a vacuum decompression concentration device to concentrate the filtrate obtained in step S7 at 45° C. This temperature can reduce the loss of heat-sensitive components.

[0031] S9. After step S8 is concentrated until there is no alcohol taste, the equipment is turned off and the extract is allowed to cool naturally to room temperature.

[0032] S10. The cooled extract in step S9 is dried using a vacuum dryer.

[0033] S11, weighing the ethanol extract of Partridge Tea obtained in step S10 to obtain 307 g of ethanol extract of Partridge Tea, and storing it in a dry, dark and cool place to maintain its stability and effectiveness.

[0034] Example 2 Effect of Partridge Tea Ethanol Extract in Preventing Obesity Induced by High-Fat and High-Cholesterol Diet in Mice In this example, the ethanol extract of Partridge Tea was used as a test object, and an experimental method of high-fat and high-cholesterol diet-induced obese mice was implemented to verify the improvement effect of the ethanol extract of Partridge Tea on lipid-lowering and weight-loss.

[0035] 1. Experimental methods: (1) Mouse grouping: Thirty-two 8-week-old male C57BL / 6J mice weighing 18-20 g were purchased and kept at a constant temperature and humidity for one week. They were divided into four groups (8 mice in each group) according to their body weight: normal control group, high-fat and high-cholesterol (HFC) model group, partridge tea extract intervention group (500 mg / kg), and positive drug rosiglitazone intervention group (10 mg / kg).

[0036] (2) Preparation of intervention drugs with ethanol extract of partridge tea: Use a precision balance to accurately weigh 200 mg of the ethanol extract of partridge tea into a 5 mL EP tube. According to the dosage of 0.4 mL / 40 g mouse, use a high-precision pipette to add 4 mL of ultrapure water to the EP tube. Vortex and sonicate to completely dissolve the ethanol extract of partridge tea, thereby preparing a 500 mg / kg dose solution.

[0037] (3) Prepare positive drugs: Use a balance with an accuracy of 1 part per 100,000 to weigh 4 mg of rosiglitazone into a 5 mL EP tube. Use a high-precision pipette to add 8 μL DMSO (2‰) and 200 μL Tween 80 (5%) to the EP tube. Vortex the solution in the EP tube to ensure that the rosiglitazone is completely dissolved. Use normal saline to make up the rest (3792 μL) to prepare a 10 mg / kg rosiglitazone solution.

[0038] (4) Drug administration to mice: The mice in the normal control group were fed with normal feed. The other groups were fed with high-fat and high-cholesterol feed (containing 60% fat and 1.2% cholesterol, purchased from Research Diet, USA). At the same time, the group treated with the extract of the quail tea extract in (1) was given the solution prepared in (2) by gavage; the group treated with the positive drug rosiglitazone was given the solution prepared in (3) by intraperitoneal injection. The drug was administered once at 10 am every day, and the weight of the mice was observed and recorded. The drug was administered continuously for 60 days.

[0039] (5) After step (4), the animals were anesthetized and blood was collected. The animals were killed by cervical dislocation and dissected. The livers were photographed and their weights were recorded. Serum was collected.

[0040] 2. Experimental results: The results are shown in Figure 2. Figure 1 It can be seen that the ethanol extract of partridge tea can effectively inhibit the weight gain of mice induced by HFC diet. During the 60-day administration, compared with the high-fat and high-cholesterol group of mice, the weight gain trend of mice in the partridge tea extract intervention group was significantly slowed down, and its intervention effect was equivalent to that of the clinical drug rosiglitazone intervention group, indicating that the ethanol extract of partridge tea can significantly improve the obesity symptoms of mice induced by a high-fat and high-cholesterol diet.

[0041] Figure 2 , 3 , 4 It can be seen that after 60 days of high-fat and high-cholesterol diet feeding, the weight of subcutaneous adipose tissue, abdominal white adipose tissue and brown adipose tissue in the model group was significantly increased compared with the control group mice (p < 0.001), and the weight of each adipose tissue of the mice was significantly reduced after intervention with the ethanol extract of partridge tea (p < 0.001).

[0042] from Figure 5 It can be seen that the expression of uncoupling protein 1 (UCP1) was reduced in the model group, and its expression was significantly increased after the intervention of quail tea extract. UCP1 is a nuclear-encoded protein expressed on the inner membrane of mitochondria, which is mainly specifically expressed in brown adipose tissue. It is a key molecule for thermogenesis in brown adipose tissue and plays an important role in thermogenesis, regulating energy metabolism and maintaining mitochondrial homeostasis. This shows that quail tea extract can promote thermogenesis in mice. In summary, quail tea extract has a positive effect on weight loss by alleviating liver damage and fat accumulation.

[0043] from Figure 6From the liver photographs, it can be seen that the liver of the normal group mice was normal in size, with smooth capsule, sharp edges, soft texture and bright red color; the liver of the model mice fed with high fat and high cholesterol was significantly enlarged, and its liver was milky white to varying degrees, with tight capsule, greasy cut surface, and obvious fatty lesions; the liver of the mice in the partridge tea extract intervention group was reduced in size, dark red in color, slightly thick capsule, and soft texture; and after rosiglitazone intervention, the liver of the mice showed obvious milky white particles with smooth edges. Pathological staining results showed that the fat droplets of each adipose tissue of the model group mice were significantly enlarged compared with the control group; compared with the model group, the fat droplet volume of the mice in the partridge tea extract group was significantly reduced after intervention.

[0044] •In summary, the mouse experiment can effectively demonstrate that the ethanol extract of partridge tea can effectively play a lipid-lowering and weight-loss role, and the ethanol extract of partridge tea has no visible tissue cytotoxicity at the effective dose.

[0045] •The above embodiments are preferred implementations of the present invention, but the implementations of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An application of quail tea ethanol extract in reducing fat and losing weight, providing a preparation method of the quail tea ethanol extract: S1, drying the quail tea material and crushing it into powder; S2, soaking it in 12% ethanol, and filtering the extract after soaking for three days, and the above operation is performed three times, and the obtained filtrate is the quail tea ethanol extract; S3, concentrating the filtrate under vacuum at 45°C to obtain an extract; further, an application of the quail tea ethanol extract, characterized in that The extract is used to treat or prevent obesity and its related symptoms caused by a high-fat and high-cholesterol diet; further, the pharmaceutical composition of the partridge tea extract is characterized in that the composition contains the partridge tea ethanol extract as an active ingredient.

2. The present invention also provides a specific application method of the ethanol extract of Partridge Tea, which specifically comprises the following steps: S1. Purchase 32 Specific Pathogen Free (SPF) grade 8-week-old and weighing 18-20 kg g male C57BL / 6J mice; S2, raising mice under constant temperature and humidity, and after one week, they were divided into 4 groups (8 mice in each group) according to their body weight: normal control group, high-fat and high-cholesterol diet model group, quail tea ethanol extract intervention group, and positive drug rosiglitazone intervention group; S3, except the normal control group, the other groups were given a high-fat diet, and the mice were given drugs by gavage / intraperitoneal injection at 10 o'clock every morning, and the weight of the mice was observed and recorded, and the drugs were given continuously for 60 days; S4, after step S3, anesthesia and blood collection were performed, and the mice were killed by cervical dislocation and dissected, and the weight of the mouse liver and adipose tissue was collected and recorded; S5, a certain mass of the mouse subcutaneous adipose tissue, abdominal white adipose tissue and brown adipose tissue collected in step S4 were weighed, paraffin sections were made, and hematoxylin-eosin (HE) staining was used for pathological section observation; S6, the brown adipose tissue paraffin sections prepared in S5 were subjected to immunohistochemical staining of UCP1, and section observation was performed; further, in step S2, the extract administration dose was 500 mg / kg, and the positive drug group was 10 mg / kg; further, in step S3, the high-fat and high-cholesterol feed formula contained 60% fat and 1.2% cholesterol.

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