Composition for inhibiting obesity as well as preparation method and application thereof

Abstract

The invention relates to the technical field of obesity inhibition, in particular to a composition for inhibiting obesity and a preparation method and application thereof. According to the invention, the red pimento extract, the suberect spatholobus stem extract and the chlamydomonas reinhardtii are combined, and the red pimento extract rich in capsaicin ester promotes body lipid oxidative metabolism, activates brown adipose tissues, promotes white adipose tissue browning and increases energy consumption by increasing body lipid metabolism heat production; meanwhile, the suberect spatholobus stem extract containing isoflavone compounds is used for maintaining the steady state of glucose and reducing insulin resistance to up-regulate metabolic regulators, chlamydomonas reinhardtii is compounded, the chlamydomonas reinhardtii has high protein, low fat, rich dietary fibers and strong satiety, weight control is facilitated, and the suberect spatholobus stem extract, the suberect spatholobus stem extract and the chlamydomonas reinhardtii have a synergistic effect; the traditional Chinese medicine composition can regulate and control metabolites, influence related signal channels, promote white fat browning, and achieve the purposes of obesity inhibition and weight loss.

Description

Technical Field

[0001] The present invention relates to the technical field of suppressing obesity, and in particular to a composition for suppressing obesity, and a preparation method and application thereof. Background Art

[0002] Obesity is a chronic metabolic disease that is caused by excessive accumulation of fat in the body and threatens health. Obesity is mainly related to factors such as diet, physique, genetics, mental state and region. With the improvement of human living standards, factors such as unreasonable diet structure, low grain intake, high intake of high-fat and high-calorie foods, and lack of physical exercise have led to an increase in overweight and obesity, which in turn has led to an increasing trend in obesity-related diseases. In addition to causing high blood pressure and diabetes, obesity is also an important risk factor for stroke.

[0003] At present, the methods for treating obesity include diet control, exercise therapy, drug therapy, surgical treatment and traditional Chinese medicine. Diet control and exercise therapy are difficult to stick to and are prone to rebound. Drugs for obesity have problems of side effects and drug resistance. Surgical treatments such as gastric bypass surgery and gastroplasty have the potential risk of malabsorption, anemia and duct stenosis. Therefore, conventional treatment methods cannot solve the problem of obesity from the root, let alone the problem of obesity rebound.

[0004] The prior art discloses a composition for weight control and its application, which improves the weight loss effect by improving intestinal health with beneficial bacteria, but ignores the regulation of adipose tissue, resulting in poor weight loss effect. Summary of the invention

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the defect that the regulation of adipose tissue is neglected in the prior art, resulting in poor weight loss effect, thereby providing a composition for inhibiting obesity and a preparation method and application thereof.

[0006] In one aspect, the present invention provides a composition for inhibiting obesity, comprising the following components: a red bell pepper extract, a Millettia reticulata extract, and Chlamydomonas reinhardtii.

[0007] The obesity-inhibiting composition provided by the present invention can promote the browning of white fat, help improve metabolism, regulate body weight, and is rich in nutrition, contains a variety of physiologically active substances, and is helpful in improving body functions.

[0008] Among them, the main active ingredient in red bell pepper extract is a class of non-irritating, low-toxic capsaicinoid substances, namely capsaicin esters. The role and mechanism of capsaicin esters in regulating glucose and lipid metabolism, anti-tumor, anti-inflammatory, enhancing exercise endurance and anti-oxidation are current research hotspots. Its biological activity in analgesia, anti-allergy, enhancing swallowing reflex, upregulating skin insulin-like growth factor levels and activating brain areas has also attracted attention. Studies have shown that capsaicin esters have the effects of increasing the body's lipid metabolism heat production, reducing body fat content, and controlling body weight. From animal studies, it can be seen that the fat-reducing effect of capsaicin esters is close to swimming and better than dieting. Many human studies have shown that capsaicin esters only promote lipid oxidation metabolism in individuals with abnormal body mass index (BMI), and do not affect the lipid metabolism balance of individuals with normal BMI. Current research generally believes that the main mechanism of capsaicin esters in regulating lipid metabolism is to activate transient receptor potential channel protein vanilloid type-1 (TRPV1). After TRPV1 is activated, it can stimulate the sympathetic nervous system and promote lipid metabolism in the body. Capsaicin esters can activate TRPV1 in the gastrointestinal tract and the sympathetic nervous system, thereby promoting adrenaline secretion and reducing body fat content. Capsaicin esters have weaker activation efficacy on TRPV1 in human embryonic kidney 293 cells than capsaicin, and the dose required to induce adrenaline and norepinephrine secretion in mice is significantly higher than capsaicin. TRPV1 is considered to be the main receptor for capsaicin esters to regulate lipid metabolism. In addition, the mechanism by which capsaicin esters promote lipid metabolism in the body is also related to its activation of brown adipose tissue (BAT) function and promotion of white adipose tissue (WAT) "browning". WAT and BAT are important tissues for energy storage and metabolic heat production in the body, respectively. Studies have shown that capsaicin esters can upregulate the transcription and expression levels of uncoupling protein (UCP) 1 in BAT and UCP2 in WAT, thereby promoting lipid oxidation and decomposition. Capsaicin esters can upregulate the expression of PR domain protein 16 in WAT, promote WAT "browning", increase the content of beige adipose tissue in the inguinal area, supraclavicular area and other parts, increase energy consumption, and inhibit the body's lipid aggregation. Capsaicin esters combined with low temperature can enhance their effects in promoting the formation of beige adipocytes and regulating lipid metabolism. In summary, capsaicin esters have the application value of safely and effectively promoting lipid metabolism and reducing body weight. The unique application of the present invention to the field of fat reduction is the strong promotion of WAT "browning" by capsaicin esters.

[0009] Millettia extract is mainly composed of isoflavone compounds (such as formononetin, daidzein, etc.), triterpenoids (such as epicorbyl alcohol, cork ketone, etc.) and steroidal compounds (β-sitosterol, carotene glycosides, campesterol, millettia alcohol). Studies have shown that Millettia extract has a significant effect on inhibiting weight gain, reducing obesity, maintaining glucose homeostasis, reducing insulin resistance and improving hepatic steatosis in diet-induced obesity (DIO) mice. In addition, the administration of Millettia has the effect of regulating the activation of brown adipose tissue (BAT) and the expression level of thermogenic genes in DIO mice. In addition, the activation of BAT treated with Millettia was also confirmed in BAT primary cells. The improvement of glucose homeostasis and insulin resistance may be related to the upregulation of MAPK and AMPK pathways in white adipose tissue (WAT) and BAT. It is worth noting that Millettia extract also has the function of improving obesity-induced intestinal flora imbalance, thereby inducing an increase in anti-obesity and anti-diabetes related bacterial genera.

[0010] Chlamydomonas reinhardtii is a single-cell eukaryotic green algae, which has been widely studied as a model organism for photosynthesis, genetics and physiology. It is rich in nutrients, including more than 40% protein, and a reasonable amino acid composition, containing eight essential amino acids for the human body; unsaturated fatty acids account for a high proportion, including ω3, ω6, ω9 fatty acids, etc.; rich in cellulose, much higher than Chlorella and Spirulina; at the same time, Chlamydomonas reinhardtii is also a high-quality source of chlorophyll, and contains a variety of essential trace elements for the human body, such as calcium, magnesium, sodium, potassium, phosphorus, selenium, zinc, etc. Chlamydomonas reinhardtii is a low glycemic index (GI) food, high crude polysaccharides help reduce the amount of monosaccharides converted into glucose, and high levels of organic selenium and niacin help maintain the normal function of insulin. Studies have found that Chlamydomonas reinhardtii has a positive effect on lowering blood sugar in animals. The crude polysaccharide content of Chlamydomonas reinhardtii is as high as 12.5%. The polysaccharide has a good lipid-lowering effect of lowering serum total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and increasing high-density lipoprotein cholesterol (HDL-C). The influence mechanism is mainly summarized as polysaccharides achieving the purpose of lowering blood lipids by affecting the absorption, distribution, metabolism, and excretion of TC and TG in the body.

[0011] The present invention creatively discovers that the combination of red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii has a synergistic effect in the technical effect of inhibiting obesity. At the same time, the combination of red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii with fishy smell can significantly reduce the fishy smell and improve the taste of the product.

[0012] In some of the embodiments, the mass ratio of red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii is (1-3.5):(4-20):(32-100).

[0013] In order to further improve the effect of suppressing obesity, the composition also includes a microecological composite preparation, which includes soluble dietary fiber and probiotics. The present invention uses a compound of probiotics and soluble dietary fiber, utilizes the regulation of multiple nutrients and metabolites, affects related signal pathways, and further promotes the browning of white fat.

[0014] In some embodiments, the soluble dietary fiber includes at least one of inulin, resistant dextrin, psyllium husk, fructooligosaccharides, xylo-oligosaccharides, galacto-oligosaccharides, and isomaltooligosaccharides.

[0015] Preferably, in the obesity-inhibiting composition, the content of inulin is 12-18wt%, the content of resistant dextrin is 8-12wt%, the content of psyllium husk is 3-8wt%, the content of fructooligosaccharide is 5-10wt%, the content of xylo-oligosaccharide is 5-10wt%, the content of galacto-oligosaccharide is 5-10wt%, and the content of isomaltooligosaccharide is 2-8wt%.

[0016] In some embodiments, the probiotics include at least one of Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium longum subspecies longum, Bifidobacterium lactis, Bifidobacterium animalis, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus paracasei.

[0017] Specifically, in the obesity-inhibiting composition, the probiotic is at least one of Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, Bifidobacterium longum subspecies longum BB536, Bifidobacterium lactis B420, Bifidobacterium animalis Bb-12, Lactobacillus plantarum Lp-115, Lactobacillus rhamnosus HN001, Bifidobacterium breve M-16V, Lactobacillus acidophilus GMNL-185, Lactobacillus rhamnosus GMNL-74, Lactobacillus plantarum GMNL-682, and Lactobacillus paracasei X11.

[0018] Preferably, in the composition for inhibiting obesity, the content of Bifidobacterium infantis is 40×10 8 ~60×10 8 CFU / 100g, the content of Bifidobacterium bifidum is 40×10 8 ~60×10 8 CFU / 100g, Bifidobacterium longum subsp. longum 50×10 8 ~60×10 8 CFU / 100g, Bifidobacterium lactis 50×10 8 ~60×10 8 CFU / 100g, Bifidobacterium animalis 20×10 8 ~40×108 CFU / 100g, Lactobacillus plantarum 10×10 8 ~30×10 8 CFU / 100g, Lactobacillus rhamnosus 40×10 8 ~60×10 8 CFU / 100g, Bifidobacterium breve 20×10 8 ~40×10 8 CFU / 100g, Lactobacillus acidophilus 40×10 8 ~60×10 8 CFU / 100g, Lactobacillus rhamnosus 20×10 8 ~40×10 8 CFU / 100g, Lactobacillus plantarum 20×10 8 ~40×10 8 CFU / 100g, Lactobacillus paracasei 10×10 8 ~20×10 8 CFU / 100g.

[0019] In another aspect, the present invention provides a method for preparing a composition for suppressing obesity, comprising mixing various raw materials according to the content of each raw material.

[0020] In some of the embodiments, the raw materials are mixed under stirring, the stirring speed is 6-10 rpm, and the stirring time is 25-40 min.

[0021] Preferably, during the stirring, the temperature is 8-20° C. and the humidity is 30-40%; and / or,

[0022] Preferably, the particle size of the composition for inhibiting obesity is 60-100 mesh.

[0023] The above preparation method is carried out in a constant temperature and humidity environment in a GMP workshop, but is not particularly limited. The solid beverage pouch packaging needs to be filled with nitrogen.

[0024] The obesity-inhibiting composition provided by the present invention or the obesity-inhibiting composition prepared by the above preparation method has any of the following applications:

[0025] (1) Preparation of drugs for preventing and / or treating obesity;

[0026] (ii) preparing drugs for regulating metabolism;

[0027] (iii) regulating the browning of white fat;

[0028] (IV) Regulate the expression of thermogenic protein-uncoupling protein 1.

[0029] Preferably, the drugs for regulating metabolism include those for regulating liver mass, total cholesterol, triglycerides and other indicators.

[0030] The technical solution of the present invention has the following advantages:

[0031] 1. The present invention provides a composition for inhibiting obesity, comprising the following components: red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii. The present invention utilizes a combination of three substances, namely, red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii. On the one hand, the red bell pepper extract rich in capsaicin esters promotes the body's lipid oxidation metabolism by increasing the body's lipid metabolism and heat production, activates brown adipose tissue, promotes the browning of white adipose tissue, increases the content of beige adipose tissue in the inguinal area, supraclavicular area and other parts, increases energy consumption, and inhibits the body's lipid aggregation. At the same time, the Millettia spatholobi extract containing isoflavone compounds is used to maintain glucose homeostasis and reduce insulin resistance to upregulate metabolic regulatory factors, such as MAPK and AMPK, etc. , and compounded with Chlamydomonas reinhardtii to further reduce the body's blood lipids. The present invention compounds red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii. Chlamydomonas reinhardtii has high protein and low fat, as well as rich dietary fiber, strong satiety, and is helpful for weight control. The three synergistically enhance the effect, regulate metabolites, and affect related signal channels, promote the browning of white fat, and achieve the purpose of inhibiting obesity and losing weight. On the other hand, the present invention uses red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii with a pungent taste to significantly reduce the fishy smell of Chlamydomonas reinhardtii and improve the taste of the product.

[0032] 2. The present invention provides a composition for suppressing obesity, further comprising a microecological composite preparation, wherein the microecological composite preparation comprises soluble dietary fiber and probiotics. The present invention utilizes soluble dietary fiber and probiotics, which can not only further adjust the taste of the composition, but also improve body function and metabolism by regulating intestinal flora and enriching nutrients, thereby achieving the purpose of suppressing obesity and losing weight.

[0033] 3. The method for preparing the composition for inhibiting obesity provided by the present invention is to mix the raw materials according to the content of each raw material. The preparation method provided by the present invention has simple process, low environmental requirements and is easy to promote. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0035] Figure 1This is a graph showing the expression of the mRNA of the thermogenic protein uncoupling protein 1 in Experimental Example 2 of the present invention;

[0036] Figure 2 is a gel electrophoresis diagram of the protein expression amount of the thermogenic protein-uncoupling protein 1 in Experimental Example 2 of the present invention;

[0037] Figure 3 This is a graph showing the weight loss results of volunteers in Experimental Example 4 of the present invention. DETAILED DESCRIPTION

[0038] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0039] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0040] Adipose tissue plays an important role in energy metabolism and endocrine organs in the human body. Adipose tissue is a tissue and organ with secretory function, which is roughly divided into white fat (WAT), brown fat (BAT) and beige fat. Adipose tissue originates from the mesoderm, which is divided into paraxial mesoderm and lateral mesoderm, producing white fat cells and brown fat cells. Brown fat cells are produced by paraxial mesoderm. White fat and brown fat have similar origins, but different development patterns and morphologies. The progenitor cells of white, brown and beige fat are mesenchymal stem cells with different molecular markers. In the presence of external stimuli, white fat and beige fat can be transformed into each other under certain conditions. In contrast, brown fat is darker in color and has a higher mitochondrial content. Beige fat is second only to brown fat, while white fat has the least mitochondria and does not produce heat. Brown fat is widely distributed throughout the body, including the interscapular region, mediastinum, kidneys, adrenal fat region, para-arterial region and neck region. The number of brown fat cells gradually decreases with age, disappearing first in the interscapular region. Brown fat exists deep in the body for a long time before disappearing to the surface area.

[0041] Compared with energy-storing white fat, brown adipose tissue can consume calories for the body by generating heat. Many studies have shown that under certain conditions, stimulating white fat can promote its transformation into brown fat. Since obesity is a metabolic disease caused by excessive lipid deposition in white fat, researchers are working to promote its energy metabolism. One way to treat metabolic diseases such as obesity is to promote the browning of white fat, and the signal transduction pathway of white fat browning may provide new ideas for the treatment of obesity and the alleviation of obesity-related glucose and lipid metabolism disorders. White fat browning is a complex process that is regulated by a variety of nutrients and metabolites and affects related signaling pathways. White fat browning is regulated by many regulatory factors, such as UCP-1, PPARγ, PRDM-16, and PGC-1α. The classic theory of "white fat browning" (WFB) is that PRDM-16 and SIRT1 induce sympathetic nervous system excitation and activation of the β3-ARs signaling pathway, exerting a deacetylation effect on PPARγ. This process depends on the induction and translocation of the transcriptional coactivator PGC1α into the nucleus and its association with the gene promoter region. In addition to sympathetic nerves, cellular energy sensing is a driving force for regulating browning by regulating transcriptional networks such as AMPK, which inhibits fat deposition and induces WFB by downregulating ACC expression and inhibiting gluconeogenesis. Therefore, in theory, it is possible to treat a variety of obesity and other metabolic disorders by inducing and activating the production of beige fat to regulate the expression of thermogenic genes.

[0042] The present invention uses red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii as main ingredients, affects relevant signal channels through nutritional supplementation and regulation of metabolites, and promotes the browning of white fat.

[0043] Among them, the main active ingredient of red bell pepper extract is a class of non-irritating, low-toxic capsaicinoids - capsaicin esters; Millettia spatholobi extract mainly contains isoflavone compounds such as formononetin, daidzein, etc., triterpenoid compounds such as epicorbyl alcohol, cork ketone, etc., and steroid compounds such as β-sitosterol, carotene glycosides, campesterol, millettia spatholobi alcohol, etc.

[0044] Capsaicin ester in red bell pepper extract is the main active ingredient, which can activate transient receptor potential channel protein vanillin subtype 1 (TRPV1). After TRPV1 is activated, it can stimulate the sympathetic nervous system and promote lipid metabolism in the body. At the same time, capsaicin ester can also activate TRPV1 in the gastrointestinal tract, further activate the sympathetic nervous system, and then promote adrenaline secretion and reduce body fat content. Furthermore, capsaicin ester promotes lipid oxidation and decomposition by upregulating the transcription and expression levels of uncoupling protein (UCP) 1 in brown adipose tissue (BAT) and UCP2 in white adipose tissue (WAT), while upregulating the expression of PR domain protein 16 in WAT, promoting WAT "browning", increasing the content of beige adipose tissue in the inguinal area, supraclavicular area and other parts, increasing energy consumption, inhibiting the body's lipid aggregation, and laying the foundation for the weight loss effect of the entire composition. When compounded with Millettia reticulata extract and Chlamydomonas reinhardtii, the role of red bell pepper extract in regulating adipose tissue metabolism can complement and enhance each other with other ingredients.

[0045] Isoflavone compounds, the main components of Millettia spatholobi extract, have significant effects on inhibiting weight gain, reducing obesity, maintaining glucose homeostasis, reducing insulin resistance and improving hepatic steatosis in diet-induced obesity (DIO) mice. In previous studies, the administration of Millettia spatholobi extract can regulate the activation of brown adipose tissue (BAT) in DIO mice and the expression levels of thermogenic genes, which was also confirmed in BAT primary cells. The improvement of glucose homeostasis and insulin resistance may be related to the upregulation of MAPK and AMPK pathways in white adipose tissue (WAT) and BAT. In addition, Millettia spatholobi extract also has the function of improving obesity-induced intestinal flora imbalance, thereby inducing an increase in anti-obesity and anti-diabetes related bacterial genera. When combined with Chlamydomonas reinhardtii and red bell pepper extracts, Millettia spatholobi extracts upregulate metabolic regulatory factors (such as MAPK and AMPK, etc.) by maintaining glucose homeostasis and reducing insulin resistance, and cooperate with the effect of red bell pepper extract in promoting lipid metabolism. It can not only stabilize glucose levels and insulin function, thereby promoting the body to better utilize energy and avoid excessive energy storage in the form of fat; at the same time, the upregulation of metabolic regulatory factors can further promote the metabolic activity of adipose tissue and enhance the effect of white fat browning. Furthermore, the effect of Millettia spatholobi extracts in improving intestinal flora imbalance also has a synergistic relationship with the effect of Chlamydomonas reinhardtii on intestinal health, jointly promoting the improvement of the intestinal microecological environment, thereby affecting the overall metabolism.

[0046] Chlamydomonas reinhardtii is a single-cell eukaryotic green algae, rich in nutrients, such as protein content greater than 40%, and a reasonable amino acid composition, containing eight essential amino acids for the human body; unsaturated fatty acids account for a high proportion, including ω3, ω6, ω9 fatty acids, etc.; rich in cellulose, much higher than Chlorella and Spirulina; it is also a high-quality source of chlorophyll, and contains a variety of trace elements necessary for the human body. Chlamydomonas reinhardtii is a low glycemic index (GI) food. High crude polysaccharides help reduce the amount of monosaccharides converted into glucose. High levels of organic selenium and niacin help maintain the normal function of insulin and have a positive effect on lowering blood sugar in animals. Its crude polysaccharide content is as high as 12.5%, which has a good lipid-lowering effect of lowering serum total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and increasing high-density lipoprotein cholesterol (HDL-C). The mechanism of influence is mainly summarized as polysaccharides affecting the absorption, distribution, metabolism, excretion and other processes of TC and TG in the body to achieve the purpose of lowering blood lipids. The high protein, low fat and rich dietary fiber characteristics of Chlamydomonas reinhardtii give it a strong sense of fullness, which helps to control weight. When combined with red bell pepper extract and Millettia spatholobi extract, the blood sugar and blood lipid lowering functions of Chlamydomonas reinhardtii complement the effects of Millettia spatholobi extract in maintaining glucose homeostasis and red bell pepper extract in regulating lipid metabolism, and jointly regulate the body's metabolic state. At the same time, the rich nutrients of Chlamydomonas reinhardtii can provide the body with necessary nutritional support, maintain body functions during the weight loss process, enable the body to better adapt to metabolic changes, and promote the browning of white fat and weight control.

[0047] The combination of red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii can not only regulate metabolites and affect related signal pathways, but also improve the intestinal flora environment, coordinate the absorption and metabolism of nutrients, enhance the intestinal barrier function, and reduce the occurrence of inflammation.

[0048] First, red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii are compounded. In terms of lipid metabolism, red bell pepper extract promotes lipid oxidation and decomposition, while Chlamydomonas reinhardtii lowers blood lipids. The two work together to regulate lipid levels in the body. In terms of glucose metabolism, Millettia spatholobi extract maintains glucose homeostasis, while Chlamydomonas reinhardtii lowers blood sugar, and they work together to maintain blood sugar balance. At the same time, they also affect related signaling pathways. Red bell pepper extract activates signaling pathways such as TRPV1 to promote lipid metabolism, Millettia spatholobi extract upregulates MAPK and AMPK pathways to regulate metabolism, and Chlamydomonas reinhardtii may affect related signaling pathways through its own nutrients. The three work together to affect the metabolism and transformation of adipose tissue, promote the browning of white fat, and achieve the purpose of inhibiting obesity and losing weight.

[0049] Secondly, in the presence of soluble dietary fiber, soluble dietary fiber can absorb water in the intestine to form a viscous substance. This environment helps to prolong the residence time of active ingredients such as capsaicin esters in red bell pepper extract in the intestine, thereby increasing their chances of binding to intestinal cell surface receptors (such as TRPV1) and better exerting their role in promoting lipid metabolism. More specifically, soluble dietary fiber creates favorable conditions for the absorption of active ingredients in Millettia reticulata extract (such as isoflavone compounds, etc.) in the intestine. The viscous environment it forms can make these active ingredients release and absorb more slowly, improve their bioavailability, and thus better play the role of maintaining glucose homeostasis and reducing insulin resistance. At the same time, soluble dietary fiber can serve as "food" for intestinal flora and promote the growth and reproduction of beneficial bacteria. In the process of metabolizing dietary fiber, beneficial bacteria will produce some short-chain fatty acids, such as acetic acid, propionic acid and butyric acid, which can affect the body's energy. For example, butyrate can promote the utilization of energy by intestinal epithelial cells and reduce the storage of energy in the form of fat, which is synergistic with the effect of red bell pepper extract in reducing body fat content; and by promoting the growth of beneficial bacteria, soluble dietary fiber can enhance the effect of Millettia reticulata extract in improving intestinal flora imbalance. The increase of beneficial bacteria can further regulate the metabolic environment in the intestine, for example, by regulating hormone secretion and signal transduction in the intestine, affecting the metabolic activities of white adipose tissue and brown adipose tissue, which cooperates with the effect of Millettia reticulata extract in regulating adipose tissue metabolism; further, some components in Chlamydomonas reinhardtii work together with dietary fiber to promote the growth of certain specific beneficial bacteria and inhibit the reproduction of harmful bacteria. This synergistic regulation of intestinal flora can improve the microecological balance of the intestine, enhance the barrier function of the intestine, reduce the absorption of intestinal endotoxins, and thus affect the overall metabolic state of the body, which is synergistic with the regulatory effect of Chlamydomonas reinhardtii itself on blood sugar and blood lipids.

[0050] Thirdly, in the presence of probiotics, after colonization in the intestine, probiotics can regulate the composition and structure of the intestinal flora. For example, Bifidobacterium infantis and Bifidobacterium bifidum can decompose some components in food and produce some metabolites. These metabolites may synergize with capsaicin esters in red bell pepper extract to further activate the sympathetic nervous system or affect signal pathways related to lipid metabolism, promote lipid metabolism in the body, and reduce body fat content. At the same time, probiotics and Millettia reticulata extract have a synergistic effect in improving intestinal flora imbalance. For example, Bifidobacterium longum subspecies longum and Bifidobacterium lactis Bacillus and other bacteria can work together with Millettia reticulata extract to increase the number of anti-obesity and anti-diabetes related bacterial genera, further improve the intestinal microecological environment, so as to better play the role of maintaining glucose homeostasis and reducing insulin resistance. More importantly, both probiotics and Chlamydomonas reinhardtii have a regulatory effect on intestinal flora. Different types of probiotics (such as Lactobacillus plantarum, Lactobacillus rhamnosus, etc.) interact with the components in Chlamydomonas reinhardtii to jointly promote the growth of beneficial bacteria, inhibit the reproduction of harmful bacteria, improve the intestinal microecological environment, and enhance the intestinal barrier function, so as to better play the role of Chlamydomonas reinhardtii in intestinal flora. Regulation of blood sugar and blood lipids; It should be noted that probiotics can enhance the barrier function of the intestine, reduce the absorption of intestinal toxins and inflammatory response, and intestinal inflammation can affect the body's metabolic state. For example, inflammatory response may lead to increased insulin resistance, fat metabolism disorders, etc. By reducing inflammatory response, probiotics can create a good internal environment for red bell pepper extract to better play its role in regulating lipid metabolism. Probiotics can interact with intestinal epithelial cells to affect the secretion and signal transduction of hormones in the intestine, thereby regulating the metabolic activities of white adipose tissue and brown adipose tissue. This regulatory effect cooperates with the effect of Millettia reticulata extract in regulating adipose tissue metabolism, and jointly promotes the browning of white fat and improves the body's metabolic level. Probiotics can affect the body's metabolic state by regulating hormone secretion and signal transduction in the intestine. It interacts with the nutrients and metabolites in Chlamydomonas reinhardtii. For example, it works together with the polysaccharides in Chlamydomonas reinhardtii to affect serum total cholesterol, triglycerides and other blood lipid indicators, and cooperates with the lipid-lowering effect of Chlamydomonas reinhardtii. At the same time, it can also affect blood sugar levels, and cooperate with the regulating effect of Chlamydomonas reinhardtii on blood sugar.

[0051] In summary, the combination of red bell pepper extract, Millettia reticulata extract and Chlamydomonas reinhardtii can more effectively achieve the goal of inhibiting obesity and weight loss. It can not only regulate the body's metabolic state, including lipid metabolism, glucose metabolism, etc., but also promote the browning of white fat, increase energy consumption, inhibit the body's lipid aggregation, and improve the intestinal microecological environment, maintain body functions, and thus achieve better weight loss effects.

[0052] In the experimental examples or comparative examples of the present invention, red bell pepper extract was purchased from Snott Biotechnology Co., Ltd., Millettia spatholobi extract was purchased from Shaanxi Hengling Natural Biological Products Co., Ltd., Chlamydomonas reinhardtii was purchased from Shanxi Touyun Biotechnology Co., Ltd., Spirulina was purchased from Hubei Yihang Biotechnology Co., Ltd., and all probiotics were commercially available.

[0053] Example 1

[0054] This embodiment provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0055] Take 2.5 g of red bell pepper extract, 5.5 g of Millettia spatholobi extract and 35 g of Chlamydomonas reinhardtii, mix them at 8 rpm for 30 min. During the mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0056] Example 2

[0057] This embodiment provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0058] Take 2.5g of red bell pepper extract, 5.5g of Millettia reticulata extract, 35g of Chlamydomonas reinhardtii, 15g of inulin, 10g of resistant dextrin, 5g of Psyllium husk, 7.5g of oligofructose, 7.5g of oligoxylose, 5g of galacto-oligosaccharide, and 5g of isomaltooligosaccharide.

[0059] And add Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, Bifidobacterium longum subsp. longum BB536, Bifidobacterium lactis B420, Bifidobacterium animalis Bb-12, Lactobacillus plantarum Lp-115, Lactobacillus rhamnosus HN001, Bifidobacterium breve M-16V, Lactobacillus acidophilus GMNL-185, Lactobacillus rhamnosus GMNL-74, Lactobacillus plantarum GMNL-682, Lactobacillus paracasei X11, wherein the content of each species is: Bifidobacterium infantis R0033 50×10 8 CFU / 100g, Bifidobacterium bifidum R0071 50×10 8 CFU / 100g, Bifidobacterium longum subsp. longum BB536 55×10 8 CFU / 100g, Bifidobacterium lactis B42055×10 8 CFU / 100g, Bifidobacterium animalis Bb-12 30×10 8 CFU / 100g, Lactobacillus plantarum Lp-11520×10 8 CFU / 100g, Lactobacillus rhamnosus HN001 50×10 8 CFU / 100g, Bifidobacterium breve M-16V30×10 8CFU / 100g, Lactobacillus acidophilus GMNL-185 50×10 8 CFU / 100g, Lactobacillus rhamnosus GMNL-74 30×10 8 CFU / 100g, Lactobacillus plantarum GMNL-682 30×10 8 CFU / 100g, Lactobacillus paracasei X11 15×10 8 CFU / 100g,

[0060] The above raw materials were mixed at a rotation speed of 8 rpm for 30 minutes. During the mixing, the ambient temperature was 15° C. and the humidity was 35%. After mixing, the mixture was passed through an 80-mesh sieve.

[0061] Example 3

[0062] This embodiment provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0063] Take 1g of red bell pepper extract, 20g of Millettia reticulata extract, 100g of Chlamydomonas reinhardtii, 30g of inulin, 20g of resistant dextrin, 12g of Psyllium husk, 12g of fructooligosaccharide, 12g of xylo-oligosaccharide, 12g of galacto-oligosaccharide, and 5g of isomaltooligosaccharide.

[0064] And add Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, Bifidobacterium longum subsp. longum BB536, Bifidobacterium lactis B420, Bifidobacterium animalis Bb-12, Lactobacillus plantarum Lp-115, Lactobacillus rhamnosus HN001, Bifidobacterium breve M-16V, Lactobacillus acidophilus GMNL-185, Lactobacillus rhamnosus GMNL-74, Lactobacillus plantarum GMNL-682, Lactobacillus paracasei X11, wherein the content of each species is: Bifidobacterium infantis R0033 40×10 8 CFU / 100g, Bifidobacterium bifidum R0071 60×10 8 CFU / 100g, Bifidobacterium longum subsp. longum BB536 50×10 8 CFU / 100g, Bifidobacterium lactis B42060×10 8 CFU / 100g, Bifidobacterium animalis Bb-12 20×10 8 CFU / 100g, Lactobacillus plantarum Lp-115 10×10 8 CFU / 100g, Lactobacillus rhamnosus HN001 60×10 8 CFU / 100g, Bifidobacterium breve M-16V20×10 8 CFU / 100g, Lactobacillus acidophilus GMNL-185 60×10 8CFU / 100g, Lactobacillus rhamnosus GMNL-74 40×10 8 CFU / 100g, Lactobacillus plantarum GMNL-682 20×10 8 CFU / 100g, Lactobacillus paracasei X11 10×10 8 CFU / 100g,

[0065] The above raw materials were mixed at a rotation speed of 6 rpm for 40 minutes. During the mixing, the ambient temperature was 20° C. and the humidity was 40%. After mixing, the mixture was passed through a 60-mesh sieve.

[0066] Example 4

[0067] This embodiment provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0068] Take 3.5g of red bell pepper extract, 4g of Millettia reticulata extract, 32g of Chlamydomonas reinhardtii, 20g of inulin, 9g of resistant dextrin, 8.5g of psyllium husk, 10g of fructooligosaccharide, 10g of xylo-oligosaccharide, 10g of galacto-oligosaccharide, and 5g of isomaltooligosaccharide.

[0069] And add Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, Bifidobacterium longum subsp. longum BB536, Bifidobacterium lactis B420, Bifidobacterium animalis Bb-12, Lactobacillus plantarum Lp-115, Lactobacillus rhamnosus HN001, Bifidobacterium breve M-16V, Lactobacillus acidophilus GMNL-185, Lactobacillus rhamnosus GMNL-74, Lactobacillus plantarum GMNL-682, Lactobacillus paracasei X11, wherein the content of each strain is: Bifidobacterium infantis R0033 60×10 8 CFU / 100g, Bifidobacterium bifidum R0071 60×10 8 CFU / 100g, Bifidobacterium longum subsp. longum BB536 60×10 8 CFU / 100g, Bifidobacterium lactis B42050×10 8 CFU / 100g, Bifidobacterium animalis Bb-12 40×10 8 CFU / 100g, Lactobacillus plantarum Lp-11530×10 8 CFU / 100g, Lactobacillus rhamnosus HN001 40×10 8 CFU / 100g, Bifidobacterium breve M-16V40×10 8 CFU / 100g, Lactobacillus acidophilus GMNL-185 40×10 8 CFU / 100g, Lactobacillus rhamnosus GMNL-74 20×10 8CFU / 100g, Lactobacillus plantarum GMNL-682 40×10 8 CFU / 100g, Lactobacillus paracasei X11 20×10 8 CFU / 100g,

[0070] The above raw materials were mixed at a rotation speed of 10 rpm for 40 minutes. During the mixing, the ambient temperature was 12° C. and the humidity was 30%. After mixing, the mixture was passed through a 100-mesh sieve.

[0071] Example 5

[0072] This embodiment provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0073] Take 2.5g of red bell pepper extract, 5.5g of Millettia reticulata extract, 34g of Chlamydomonas reinhardtii, 14g of inulin, 14g of resistant dextrin, 3.5g of Psyllium husk, 11g of fructooligosaccharide, 11g of xylo-oligosaccharide, 11g of galacto-oligosaccharide, and 8.5g of isomaltooligosaccharide.

[0074] And add Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, Bifidobacterium longum subsp. longum BB536, Bifidobacterium lactis B420, Bifidobacterium animalis Bb-12, Lactobacillus plantarum Lp-115, Lactobacillus rhamnosus HN001, Bifidobacterium breve M-16V, Lactobacillus acidophilus GMNL-185, Lactobacillus rhamnosus GMNL-74, Lactobacillus plantarum GMNL-682, Lactobacillus paracasei X11, wherein the content of each species is: Bifidobacterium infantis R0033 50×10 8 CFU / 100g, Bifidobacterium bifidum R0071 50×10 8 CFU / 100g, Bifidobacterium longum subsp. longum BB536 55×10 8 CFU / 100g, Bifidobacterium lactis B42055×10 8 CFU / 100g, Bifidobacterium animalis Bb-12 30×10 8 CFU / 100g, Lactobacillus plantarum Lp-11520×10 8 CFU / 100g, Lactobacillus rhamnosus HN001 50×10 8 CFU / 100g, Bifidobacterium breve M-16V30×10 8 CFU / 100g, Lactobacillus acidophilus GMNL-185 50×10 8 CFU / 100g, Lactobacillus rhamnosus GMNL-74 30×10 8 CFU / 100g, Lactobacillus plantarum GMNL-682 30×10 8CFU / 100g, Lactobacillus paracasei X11 15×10 8 CFU / 100g,

[0075] The above raw materials were mixed at a rotation speed of 10 rpm for 25 minutes. During the mixing, the ambient temperature was 8° C. and the humidity was 35%. After mixing, the mixture was passed through an 80-mesh sieve.

[0076] Comparative Example 1

[0077] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0078] Take 41 g of red bell pepper extract and mix it at 8 rpm for 30 minutes. During mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0079] Comparative Example 2

[0080] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0081] Take 41 g of Millettia spatholobi extract and mix it at 8 rpm for 30 min. During mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0082] Comparative Example 3

[0083] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0084] Take 41 g of Chlamydomonas reinhardtii and mix it at 8 rpm for 30 min. During the mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0085] Comparative Example 4

[0086] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0087] Take 17.1 g of red bell pepper extract and 23.9 g of Millettia spatholobi extract, mix them at 8 rpm for 30 min. During the mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0088] Comparative Example 5

[0089] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0090] Take 2.73 g of red bell pepper extract and 38.27 g of Chlamydomonas reinhardtii, mix them at 8 rpm for 30 min. During the mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0091] Comparative Example 6

[0092] This comparative example provides a method for preparing a composition for inhibiting obesity, and the specific steps and parameters are as follows:

[0093] Take 3.72 g of Millettia spatholobi extract and 37.27 g of Chlamydomonas reinhardtii, mix them at 8 rpm for 30 min. During the mixing, the ambient temperature is 15° C. and the humidity is 35%. After mixing, pass the mixture through an 80-mesh sieve.

[0094] Comparative Example 7

[0095] This comparative example provides a method for preparing a composition for inhibiting obesity. The specific steps and parameters are the same as those in Example 1, except that an equal mass of Spirulina is used to replace Chlamydomonas reinhardtii.

[0096] Experimental Example 1

[0097] 5-week-old male C57BL / 6N mice were housed in individual cages in an environmental temperature of 22±2°C, a humidity of 55%±3%, a light-dark cycle of 12h, free access to water and food, and weighed once a day. After one week of adaptive feeding, they were fed with a high-fat diet (HFD) for 8 consecutive weeks. After the eighth week of high-fat feeding, the body weight and fasting blood glucose of the mice were tested. The body weight of the mice was 30% heavier than that of normal mice, and the fasting blood glucose was greater than or equal to 7mmol / L, indicating that the obese mouse model was successfully established. Among them, the high-fat diet was 60kcal% fat, purchased from Yushu Biotechnology (Shanghai) Co., Ltd., and the conventional feed formula was 33.6wt%, lard 26wt%, sucrose 8wt%, casein 13.5wt%, whole milk powder 10.8wt%, calcium hydrogen phosphate 2wt%, calcium carbonate 1.2wt%, microcrystalline cellulose 1.9wt%, and mixed trace elements (vitamins, minerals) 3wt%.

[0098] 132 successfully modeled obese mice were randomly divided into 11 groups, namely blank control group, high-fat diet group, and experimental groups 1-9, with 12 mice in each group.

[0099] The blank control group was fed with conventional feed;

[0100] The high-fat diet group was fed a high-fat diet;

[0101] The experimental groups 1-9 were fed freely with HFD high-fat feed, and at the same time, 1 mL of the aqueous solution of the composition for suppressing obesity prepared in Examples 1-2 and Comparative Examples 1-7 was gavaged sequentially, wherein the concentration of the aqueous solution of the composition for suppressing obesity prepared in Examples or Comparative Examples in the experimental groups 1-9 was 100 mg / mL.

[0102] After 10 weeks of feeding according to different treatments, the mice in each group were fasted for 12 hours, and then their body weights were measured. The mice were anesthetized with ether, and the abdomen was opened to collect blood from the heart and liver tissue. The blood was centrifuged at low temperature to obtain the supernatant (serum), which was placed in a 4°C refrigerator for later use; the liver was washed 3 times with saline, wiped dry and weighed.

[0103] ELISA kits were used to measure serum total cholesterol (the kit was purchased from Abcam, USA), triglycerides (the kit was purchased from Abbkine, USA), and fasting glucose (the kit was purchased from Abcam, USA). The results are shown in Table 1.

[0104] Table 1 Physical index parameters of mice in each group

[0105]

[0106]

[0107] In Table 1, * indicates that compared with the high-fat diet group, P < 0.05, # indicates that compared with experimental group 1, P < 0.05

[0108] According to the data in Table 1, compared with the high-fat diet group, the compositions for inhibiting obesity provided in Examples 1 and 2 of the present invention can significantly reduce the weight of mice, and at the same time significantly reduce liver mass, total cholesterol, triglycerides, and fasting blood sugar. Although the compositions prepared in Comparative Examples 1-7 in Experimental Groups 3-9 can also reduce the weight of mice to a certain extent, the effect is not as good as the compositions provided in the experimental examples of the present invention.

[0109] Experimental Example 2

[0110] Diabetes caused by obesity is usually accompanied by hyperglycemia and hyperlipidemia. Its pathogenesis is: when the body is obese, adipose tissue releases excessive fatty acids or lipids, and long-term lipid accumulation causes insulin resistance. Adipose tissue heat production is an effective way to improve obesity-induced insulin resistance. Brown adipose tissue can express a large amount of thermogenic protein-Uncoupling protein 1 (UCP1), causing the body to produce heat and improve insulin resistance. White adipose tissue (WAT), whose function is to store lipids, has very little mitochondrial content, but white adipose tissue can also express UCP1 after browning, promoting heat production. Therefore, UCP1 is a representative expression protein of brown adipose tissue, and increased expression of UCP1 levels can also be regarded as a representative sign of white adipose browning.

[0111] This experimental example verifies that the obesity-suppressing composition of the present invention has the function of promoting the browning of white fat by detecting the expression level of UCP1.

[0112] 1. Verification of UCP1 mRNA expression:

[0113] 1.1 Experimental process:

[0114] Twenty-four 6-week-old SD male rats were randomly divided into three groups, 8 rats in each group, namely the control group, the high-fat feeding group and the experimental group.

[0115] The rats in the control group were fed a basic diet (protein 22wt%, fat 12wt%, carbohydrate 66wt%) throughout the whole process.

[0116] The high-fat feeding group was fed a high-fat diet (protein 9wt%, fat 66wt%, carbohydrate 25wt%) throughout the whole process.

[0117] After the experimental group was fed with a high-fat diet for 6 weeks, in addition to the high-fat diet, 800 mg / kg of the obesity-inhibiting composition prepared in Example 1 was intragastrically administered to each mouse every day;

[0118] The mice in each group were fed according to the above treatment for 24 weeks, and then killed by decapitation after ether anesthesia. The adipose tissue of the scapula was obtained and stored at -80°C for subsequent research.

[0119] 1.2、UCP-1mRNA expression:

[0120] The preserved brown fat was minced and fully homogenized by adding 1 mL of total RNA extraction reagent (TRIzol). Total RNA was extracted according to the instructions of the QIAGEN kit (Qiagen, RNA Mini-Extraction Kit, Qiagen, 74104), and the RNA concentration was determined using Nanodrop ND1000.

[0121] cDNA was synthesized according to the instructions of the reverse transcription kit (Biosystems). Each reaction system contained 1 μg RNA, 2 μL 10× RT buffer, 0.8 μL dNTP, 2 μL random primers, 1 μL reverse transcriptase, and the rest was ddH2O. The total reaction system was 20 μL. The reaction conditions were 25°C for 10 min, 37°C for 120 min, and 85°C for 5 min. The samples were taken out after 4°C and stored at -20°C.

[0122] The total PCR reaction system was 10 μL, including 2 μL of cDNA template, 5 μL of TaqMan@Master, 0.5 μL of primers (sequences shown in Table 2), and the rest was ddH2O.

[0123] The PCR reaction conditions were 95°C for 10 min, 95°C for 10 s, 60°C for 1 min, 72°C for 20 s, 40 cycles, and 72°C for 5 min. The reaction was repeated 3 times, and GAPDH was used as the internal reference gene. The relative expression level of the target gene was calculated using the formula 2^(△(△CT)).

[0124] Table 2 PCR primer information

[0125] Gene Chain of Justice Antisense strand GAPDH 5'-GTATGACTCTACCCACGGCAAAT-3' 5'-TTCCCGTTGATGACCAGCTT-3' UCP-1 5'-TGGCGTGGCGGTATTCA-3' 5'-TGGCTTTGTGCTTGCATTCT-3'

[0126] Test results are shown in Figure 1 It can be seen that the obesity-inhibiting composition provided by the embodiment of the present invention can increase the expression level of UCP-1 mRNA.

[0127] 2. Verification of UCP-1 protein expression:

[0128] Take the brown fat of each group of mice obtained in Experimental Example 1.1, add 1 mL of RIPA lysis buffer, and after fully homogenizing, use the BCA method to extract total protein;

[0129] 40 μg of protein sample was added to 10% SDS-PAGE spotting wells, electrophoresed at 100 V for 2 h, gel transferred to PVDF membrane, blocked with 5% skim milk, incubated with UCP-1 and β-actin primary antibodies (purchased from Santa Cruz) at 4°C overnight, washed 3 times with TBST, added with horseradish peroxidase (HRP) enzyme-labeled secondary antibody (purchased from Santa Cruz) at room temperature for 30 min, washed 3 times with TBST buffer; developed and fixed by chemiluminescence, and the film was scanned and photographed. The gel electrophoresis diagram is shown in Figure 2 It can be seen that the obesity-inhibiting composition provided by the embodiment of the present invention can increase the expression level of UCP-1 protein.

[0130] Experimental Example 3

[0131] Thirty-six 5-week-old Kunming mice (male) with the same physiological conditions (30 days old) were studied. The initial body weight was 20-24 g, and the experiment was started after the experimental animals were adaptively fed for 5 days.

[0132] Mice were fed with a high-fat diet for 30 days (the formula of the high-fat diet was 10% lard, 1% cholesterol, and 89% basic feed) to establish a hyperlipidemia model.

[0133] The 36 mice with hyperlipidemia models were randomly divided into 3 groups, and the weight of each group of mice before intervention was recorded. The groups were blank control group, experimental group 1, and experimental group 2. The experimental group 1 was intragastrically administered with 100 mg of the obesity-inhibiting composition prepared in Example 1 per kg of mice every day (the concentration of the obesity-inhibiting composition aqueous solution was 100 mg / mL), and the experimental group 2 was intragastrically administered with 200 mg of the obesity-inhibiting composition prepared in Example 1 per kg of mice every day (the concentration of the obesity-inhibiting composition aqueous solution was 200 mg / mL). The blank control group was gavaged with an equal volume of normal saline for 21 consecutive days (the corresponding dietary pattern was maintained during the gavage, that is, the blank control group and each experimental group still ate high-fat feed freely). After recording the body weight of each group of mice, they were anesthetized with ether and then decapitated. The adipose tissue of the scapula was fixed in formaldehyde calcium solution (10 ml formaldehyde and 1 g anhydrous calcium chloride to make 100 ml solution) for pathological histological examination, in which the size of the blank area after HE staining of scapular brown fat represented the fat content, and a semi-quantitative analysis was performed.

[0134] The body weight records and brown fat percentage of mice in each group are shown in Table 3.

[0135] Table 3 Test results of mice in each group

[0136] Weight before intervention Body weight after intervention Brown fat blank area% Blank control group 22.78±1.82 34.58±1.91 85.37±4.19 Experimental Group 1 22.23±1.13 31.31±2.34* 75.27±3.94* Experimental Group 2 22.39±1.75 29.67±2.51* 61.47±3.07*

[0137] *P<0.05 compared with the blank control group

[0138] According to Table 3, after 30 days of high-fat feeding, the comparison before and after intervention was obtained (P<0.05); the weight of mice in the high-concentration group was lower than that of mice in the low-concentration group; the weight of mice in the low-concentration group was lower than that of the control group, and there was a statistical difference (P<0.05). This shows that the higher the concentration of the composition that inhibits obesity, the better the effect of obesity resistance.

[0139] After gavage of different concentrations of the composition for suppressing obesity in three groups of mice, the blank area size after HE staining of scapular brown fat was semi-quantitatively analyzed to represent the fat content. Compared with the control group, there were statistical differences between the low-concentration group and the high-concentration group (P<0.05). It was proved that the composition for suppressing obesity of the present invention can significantly reduce the blank area in BAT tissue and promote BAT metabolism.

[0140] Experimental Example 4

[0141] A total of 189 obese volunteers were included from May 2023 to April 2024. Aged 32-66 years old, including 98 females and 91 males, with a BMI between 24.1 and 33.4, and the volunteers were randomly divided into three groups, namely, an experimental group, a lifestyle control group, and a blank control group. The experimental group had a normal diet for breakfast, lunch, and dinner, and the amount of food intake was freely adjusted according to changes in their own appetite. Physical exercise was avoided or reduced as much as possible, and daily activities were not restricted. Dinner was an additional 20g of the composition for inhibiting obesity prepared in Example 3; the lifestyle control group had a total daily dietary intake of no more than 1500kcal, exercised for 30min a day, and the blank control group had a normal diet without any intervention.

[0142] After 13 weeks, the body weight of volunteers in each group was measured. The results are shown in Table 4 and Figure 3 .

[0143] Table 4 Weight loss results

[0144] Experimental Group Lifestyle control group Blank control group Decrease ≥ 5% 98.43％#* 26.56％ 3.27％ Decrease ≥ 10% 82.32％#* 10.93％ 0％ Decrease ≥ 15% 48.43％#* 7.81％ 0％ Decrease ≥ 20% 15.62％#* 1.56％ 0％

[0145] #P<0.001 compared with lifestyle intervention, *P<0.001 compared with blank control group

[0146] According to Table 4 and Figure 3 It can be seen that, compared with the lifestyle intervention control group and the blank control group, the obesity-inhibiting composition provided by the embodiment of the present invention can significantly reduce body weight, and there is a very significant statistical difference.

[0147] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A composition for inhibiting obesity, characterized in that: The invention comprises the following ingredients: red bell pepper extract, Millettia reticulata extract and Chlamydomonas reinhardtii.

2. The composition for suppressing obesity according to claim 1, characterized in that: The mass ratio of red bell pepper extract, Millettia spatholobi extract and Chlamydomonas reinhardtii is (1-3.5):(4-20):(32-100).

3. The composition for suppressing obesity according to claim 2, characterized in that: It also includes a microecological composite preparation, which includes soluble dietary fiber and probiotics.

4. The composition for suppressing obesity according to claim 3, characterized in that: The soluble dietary fiber includes at least one of inulin, resistant dextrin, psyllium husk, fructooligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide and isomaltooligosaccharide.

5. The composition for suppressing obesity according to claim 4, characterized in that: The probiotics include at least one of Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium longum subspecies longum, Bifidobacterium lactis, Bifidobacterium animalis, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus paracasei.

6. The composition for suppressing obesity according to claim 5, characterized in that: In the composition for inhibiting obesity, the content of inulin is 12-18 wt %; and / or, The content of the resistant dextrin is 8-12wt%; and / or, The content of psyllium husk is 3-8wt%; and / or, The content of oligofructose is 5-10wt%; and / or, The content of xylooligosaccharide is 5-10wt%; and / or, The content of the galacto-oligosaccharide is 5-10wt%; and / or, The content of isomaltooligosaccharide is 2-8wt%.

7. The composition for suppressing obesity according to claim 6, characterized in that: In the composition for inhibiting obesity, the content of infant Bifidobacterium is 40×10 8 ~60×10 8 CFU / 100g; and / or, The content of Bifidobacterium bifidum is 40×10 8 ~60×10 8 CFU / 100g; and / or, Bifidobacterium longum subsp. longum 50×10 8 ~60×10 8 CFU / 100g; and / or, Bifidobacterium lactis 50×10 8 ~60×10 8 CFU / 100g; and / or, Bifidobacterium animalis 20×10 8 ~40×10 8 CFU / 100g; and / or, Lactobacillus plantarum 10×10 8 ~30×10 8 CFU / 100g; and / or, Lactobacillus rhamnosus 40×10 8 ~60×10 8 CFU / 100g; and / or, Bifidobacterium breve 20×10 8 ~40×10 8 CFU / 100g; and / or, Lactobacillus acidophilus 40×10 8 ~60×10 8 CFU / 100g; and / or, Lactobacillus rhamnosus 20×10 8 ~40×10 8 CFU / 100g; and / or, Lactobacillus plantarum 20×10 8 ~40×10 8 CFU / 100g; and / or, Lactobacillus paracasei 10×10 8 ~20×10 8 CFU / 100g.

8. A method for preparing the composition for suppressing obesity according to any one of claims 1 to 7, characterized in that: Mix the raw materials according to their contents.

9. The preparation method according to claim 8, characterized in that: The raw materials are mixed under stirring conditions, the stirring speed is 6-10 rpm, and the stirring time is 25-40 min; and / or, When the raw materials are stirred, the temperature is 8-20°C and the humidity is 30-40%; and / or, The particle size of the composition for inhibiting obesity is 60-100 meshes.

10. An anti-obesity composition according to any one of claims 1 to 7 or an anti-obesity composition prepared by the method for preparing the anti-obesity composition according to any one of claims 8 to 9 having any of the following uses: (1) Preparation of drugs for preventing and / or treating obesity; (ii) preparing drugs for regulating metabolism; (iii) regulating the browning of white fat; (IV) Regulate the expression of thermogenic protein-uncoupling protein 1.

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