Application of cooperation of Brawsonia globosa and traditional Chinese medicine extracting solution in preparation of weight-reducing and lipid-lowering medicine or food

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to an application of a Blauraceae globoid synergistic traditional Chinese medicine extracting solution in preparation of a weight-reducing and lipid-lowering medicine or food, and the medicine or food has any one of the following functions: (a) weight reduction; (b) lowering lipid; the traditional Chinese medicine extracting solution is a mulberry leaf extracting solution and a coix seed extracting solution, the preservation number of the Blauraceae is ATCC 29236, and the name of the Blauraceae is Blauraceae. According to the invention, the synergistic effect of the Brawsonia globosa and the mulberry leaf and coix seed extract can play a significant and efficient role in reducing weight and lipid, the effect is significantly better than that of single use of traditional Chinese medicines or single use of bacterial liquid, and the synergistic effect is played; in addition, compared with the single use of the mulberry leaf and coix seed extracting solution, the synergistic effect of the Brawsonia globosa and the mulberry leaf and coix seed extracting solution has a more stable effect in the aspect of reducing weight and lipid; in addition, natural strains and traditional Chinese medicine extracts are adopted, safety is high, and the method is suitable for preparing functional food or medicine.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and specifically relates to the application of Blautia coccoides in combination with traditional Chinese medicine extract in the preparation of weight loss and lipid-lowering drugs or foods. Background Art

[0002] Existing studies have shown that traditional Chinese medicine extracts such as coix seed extract and mulberry leaf extract can play a role in reducing lipid and losing weight. Among them, coix seed extract can reduce the levels of triglycerides, total cholesterol, and leptin in serum, regulate the expression of leptin and TNF-α, reduce the volume of white adipose tissue and the blood lipid content in serum; mulberry leaf water extract can reduce Lee's index, improve blood lipid levels, and relieve fatty liver lesions. Moreover, the flavonoid substances in mulberry leaf extract can reduce sugar absorption by inhibiting α-glucosidase. When used for reducing lipid and losing weight, although these two substances belong to traditional food and medicine homology substances with low toxicity and side effects, when traditional Chinese medicine extracts such as mulberry leaf and coix seed are used alone, the bioavailability is relatively low, it is difficult to fully exert their lipid-lowering and hypoglycemic effects, and high doses or long-term use are required. It is also difficult to regulate metabolism through multiple targets.

[0003] There are also studies that attempt to use microorganisms for reducing lipid and losing weight. For example, CN114231472A discloses a Lactobacillus probiotic for the preparation of lipid-lowering drugs, which can reduce total cholesterol, triglycerides, and low-density lipoprotein in human serum and increase high-density lipoprotein; another example is that CN118787668A discloses a Ligilactobacillus salivarius for the treatment of obesity, which can inhibit weight gain, reduce the area of white adipocytes, inhibit the differentiation of preadipocytes into adipocytes, reduce lipid accumulation in liver cells, and improve blood lipid levels, etc.; and another example is that CN119280285A discloses a Lacticaseibacillus rhamnosus NKU FL1-8 or its bacterial agent for preventing or treating obesity caused by high-fat diet, which can reduce the increase in adipose tissue weight and reduce body weight. Although the strains show certain intervention effects, the existing technologies mostly focus on single strains, and the intervention effects of single strains may fluctuate due to differences in individual intestinal environments. For example, existing probiotics such as Bifidobacterium and Lactobacillus have limited effects in regulating intestinal flora and improving metabolism, and are easily affected by the intestinal environment with insufficient stability, and it is necessary to maintain the balance of the flora for a long time. Summary of the Invention

[0004] The purpose of the present invention is to provide the application of Blautia coccoides in combination with traditional Chinese medicine extract in weight loss and lipid reduction to solve the above problems. Through the synergistic effect of the strain and traditional Chinese medicine, the present invention significantly enhances the effects of weight loss, lipid reduction, and metabolism improvement.

[0005] According to one aspect of the present invention, there is provided the application of Blautia coccoides in combination with traditional Chinese medicine extract in weight loss and lipid reduction, and the drug or food has any one of the following functions:

[0006] (a) Weight loss;

[0007] (b) Lipid reduction;

[0008] The traditional Chinese medicine extract is the mulberry leaf extract and the coix seed extract. The preservation number of Blautia coccoides is ATCC29236, and the name is Blautia coccoides.

[0009] In some embodiments, the mulberry leaf extract is prepared by the following method:

[0010] (1) Put the mulberry leaves into water, decoct and filter to obtain the first filter residue and the first filtrate;

[0011] (2) Add water to the first filter residue, decoct again and filter to obtain the second filtrate;

[0012] (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain.

[0013] Preferably, the mulberry leaf extract is prepared by the following method:

[0014] (1) Put the mulberry leaves into 8 to 12 times the weight of purified water, decoct for 0.6 to 2 hours and filter to obtain the first filter residue and the first filtrate;

[0015] (2) Add 6 to 10 times the weight of purified water to the first filter residue, decoct again for 0.3 to 1 hour and filter to obtain the second filtrate;

[0016] (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain.

[0017] In some embodiments, the coix seed extract is prepared by the following method:

[0018] (1) Put the soaked coix seeds into water, decoct and filter to obtain the first filter residue and the first filtrate;

[0019] (2) Add water to the first filter residue, decoct again and filter to obtain the second filtrate;

[0020] (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain.

[0021] Preferably, the coix seed extract is prepared by the following method:

[0022] (1) Soak the coix seeds for 0.3 to 1 hour, put the soaked coix seeds into 8 to 12 times the weight of purified water, decoct for 0.6 to 2 hours and filter to obtain the first filter residue and the first filtrate;

[0023] (2) Add purified water with a weight 6 to 10 times that of the first filter residue, decoct again for 0.3 to 1 hour and then filter to obtain the second filtrate;

[0024] (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain the product.

[0025] In some embodiments, Blautia coccoides needs to be activated through the following steps before use:

[0026] Inoculate Blautia coccoides into the modified chopped meat medium of ATCC 1490, culture under anaerobic conditions at 36°C to 37°C for 48 to 72 hours, then detect the bacterial liquid concentration, and adjust the bacterial liquid concentration to 1×10 8 CFU / mL to obtain the activated Blautia coccoides bacterial liquid.

[0027] In some embodiments, the volume ratio of the Blautia coccoides bacterial liquid to the traditional Chinese medicine extract in the drug or food is 1:2 to 4. Preferably, the volume ratio of the Blautia coccoides bacterial liquid to the traditional Chinese medicine extract is 1:3.

[0028] In some embodiments, lipid reduction includes reducing at least one of low-density lipoprotein, subcutaneous fat, perirenal fat, and epididymal fat.

[0029] According to another aspect of the present invention, there is provided a weight loss and lipid reduction drug, the active ingredients of which are Blautia coccoides bacterial liquid and traditional Chinese medicine extract, and the volume ratio of the two is 1:3; the traditional Chinese medicine extract is mulberry leaf extract and coix seed extract, and the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.36 g / mL to 0.5 g / mL; the concentration of the Blautia coccoides bacterial liquid is 1×10 8 CFU / mL. Preferably, the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.48 g / mL to 0.5 g / mL.

[0030] According to still another aspect of the present invention, there is provided a functional food for weight loss and lipid reduction, the active ingredients of which are Blautia coccoides bacterial liquid and traditional Chinese medicine extract, and the volume ratio of the two is 1:3; the traditional Chinese medicine extract is mulberry leaf extract and coix seed extract, and the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.36 g / mL to 0.5 g / mL; the concentration of the Blautia coccoides bacterial liquid is 1×10 8 CFU / mL. Preferably, the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.48 g / mL to 0.5 g / mL

[0031] The technical effect of the present invention is that Blautia coccoides and the extract of mulberry leaves and coix seeds can play a significant and efficient role in weight loss and lipid reduction, and the effect is significantly better than using traditional Chinese medicine alone or using the bacterial solution alone, showing a synergistic effect; moreover, compared with using the extract of mulberry leaves and coix seeds alone, the combination of Blautia coccoides and the extract of mulberry leaves and coix seeds has a more stable effect in weight loss and lipid reduction; in addition, the present invention uses natural strains and traditional Chinese medicine extracts, which are highly safe and suitable for preparing functional foods or drugs. Brief Description of the Drawings

[0032] Figure 1 It is the composition of the top 20 metabolites in the relative quantification of the macro-target metabolome of the mixture of the bacterial solution and the traditional Chinese medicine extract.

[0033] Figures 2 - 3 It is the comparison of the body weight changes of six groups of experimental mice after intervention from 0 to 18 weeks in the animal experiment of the present invention.

[0034] Figure 4 It is the comparison of the levels of low-density lipoprotein cholesterol (LDL-C) of six groups of experimental mice after 18 weeks of intervention in the animal experiment of the present invention.

[0035] Figure 5 It is the comparison of the levels of subcutaneous fat of six groups of experimental mice after 18 weeks of intervention in the animal experiment of the present invention.

[0036] Figure 6 It is the comparison of the levels of perirenal fat of six groups of experimental mice after 18 weeks of intervention in the animal experiment of the present invention.

[0037] Figure 7 It is the comparison of the levels of epididymal fat of six groups of experimental mice after 18 weeks of intervention in the animal experiment of the present invention.

[0038] Figure 8 It is the stained section images of liver cells and subcutaneous fat cells of the normal control group (ND) mice in the animal experiment of the present invention.

[0039] Figure 9 It is the stained section images of liver cells and subcutaneous fat cells of the high-fat control group (HFD) mice in the animal experiment of the present invention.

[0040] Figure 10 It is the stained section images of liver cells and subcutaneous fat cells of the group of mice treated with the bacterial solution combined with 2X traditional Chinese medicine (HFD_BC_Extract_2X) in the animal experiment of the present invention. Detailed Embodiments

[0041] The present invention will be further described in detail below in conjunction with specific embodiments. Unless otherwise specified, the raw materials and reagents used in the examples are all commercially available. Among them, the Blautia coccoides strain was purchased from Ningbo Mingzhou Biotechnology Co., Ltd. It should be noted that in the table, SD represents the standard deviation, CV represents the coefficient of variation, IQR represents the interquartile range, Ranger represents the range, and MAD represents the median absolute deviation. These parameters are indicators used to reflect data fluctuations and stability.

[0042] Instruments used: biological safety cabinet (model 1389), biochemical incubator (model SPX-250BSH-II), vertical pressure steam sterilizer (model YXQ-75FII), sealed culture tank (model 7.0L), bacterial turbidimeter (model WGZ-XT), low-temperature high-speed centrifuge (model 5810R), small refrigerated centrifuge (model Heraeus Fresco 21), YP20002B weighing scale (manufacturer: Shanghai Hengji Scientific Instruments Co., Ltd.), N-1300 rotary evaporator (manufacturer: Shanghai Ailang Instruments Co., Ltd.), ACS-D11 electronic scale (manufacturer: Shanghai Qianfeng Electronic Instruments Co., Ltd.).

[0043] Reagents used: ATCC medium: 1490 modified cooked meat medium (manufacturer: Qingdao Haibo Biotechnology Co., Ltd.), vitamin K1 solution (manufacturer: Ningbo Mingzhou Biotechnology Co., Ltd.), sodium chloride erythromycin (manufacturer: Ningbo Mingzhou Biotechnology Co., Ltd.), methanol (manufacturer: Merck).

[0044] I. Preparation of bacterial liquid

[0045] 1. Inoculate Blautia coccoides (preservation number: ATCC 29236) into ATCC 1490 modified cooked meat medium and culture it under anaerobic conditions at 36 °C for 48 - 72 h until OD 600 = 1.0 - 1.5.

[0046] 2. Use a bacterial turbidimeter to detect the concentration of the bacterial liquid and adjust it to 1×10 8 CFU / mL.

[0047] II. Preparation of traditional Chinese medicine extract

[0048] 1. Mulberry leaf extract

[0049] (1) Take 1128 g of mulberry leaves, add 10 times the amount of purified water, decoct for 1 hour, and filter to obtain the first filtrate and the first filter residue;

[0050] (2) Add 8 times the amount of purified water to the first filter residue, decoct for 0.5 h, and filter to obtain the second filtrate;

[0051] (3) Combine the first filtrate and the second filtrate, and concentrate to a crude drug concentration of 0.48 g / mL (high dose 2X) or 0.24 g / mL (low dose 1X).

[0052] 2. Coix Seed Extract

[0053] (1) Take 1128 g of coix seeds, soak for 0.5 hours, add 10 times the amount of purified water, decoct for 1 hour, and filter to obtain the first filtrate and the first residue.

[0054] (2) Add 8 times the amount of purified water to the first residue, decoct for 0.5 hours, and filter.

[0055] (2) Combine the first filtrate and the second filtrate, and concentrate to a crude drug concentration of 0.48 g / mL (high dose 2X) or 0.24 g / mL (low dose 1X).

[0056] III. Synergistic Effect Verification Experiment

[0057] 1. Animal Experiment

[0058] Select 24 male C57BL / 6J mice at 4 - 5 weeks old. After the quarantine, randomly divide them into 6 groups according to body weight, with 4 mice in each group, namely normal control group (ND), high-fat control group (HFD), bacterial solution group (HFD_BC_Activated), 2X traditional Chinese medicine group (HFD_Extract_2X), bacterial solution combined with 1X traditional Chinese medicine group (HFD_BC_Extract_1X), and bacterial solution combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X).

[0059] Construction of obese mouse model: Except for the normal control group (ND) with normal diet, the other groups are fed a high-fat diet with 60% of energy from fat.

[0060] Intervention operation: Starting from the model establishment, the bacterial solution group (HFD_BC_Activated), 2X traditional Chinese medicine group (HFD_Extract_2X), bacterial solution combined with 1X traditional Chinese medicine group (HFD_BC_Extract_1X), and bacterial solution combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) are given 200 μL of the corresponding drug by gavage, with a bacterial solution concentration of 1×10 8 CFU / ml. The normal control group (ND) and the high-fat control group (HFD) are given an equal amount of fresh medium, and gavage is performed once every other day for 18 weeks; record body weight and blood glucose every week. After the experiment, measure the weights of liver tissue and adipose tissue and the four blood lipids (TG, TC, LDL-C, HDL-C).

[0061] After the experiment, the liver and adipose tissues of mice in the normal control group (ND), high-fat control group (HFD), and the group treated with a combination of bacterial liquid and 2X traditional Chinese medicine (HFD_BC_Extract_2X) were stained.

[0062] 2. Test Results

[0063] The macro-target metabolomic characteristics of the mixture of bacterial liquid and traditional Chinese medicine extract are as Figure 1 shown in Table 1, and OD 600 = 1.0 - 1.5.

[0064] Table 1 Composition of the top 20 metabolites in the relative quantification of the macro-target metabolome of the mixture of bacterial liquid and traditional Chinese medicine extract

[0065]

[0066]

[0067] Figure 2 , Figure 3 Tables 2 and 3 show the weight changes and comparisons of six groups of mice in the animal experiment of the present invention from 0 to 18 weeks after intervention. Taking the mice in the high-fat control group (HFD) as a reference, the weights of the 2X traditional Chinese medicine group (HFD_Ex tract_2X) (p = 0.014) and the group treated with a combination of bacterial liquid and 2X traditional Chinese medicine (HFD_BC_Extract_2X) (p = 0.014) were significantly lower than those of the reference group, indicating that the 2X traditional Chinese medicine group and the group treated with a combination of bacterial liquid and 2X traditional Chinese medicine exerted a significant weight loss effect.

[0068] Table 2 Weight changes of mice in different groups from 0 to 18 weeks after intervention

[0069]

[0070] Table 3 Comparison of the weights of mice in different groups at 18 weeks after intervention

[0071] Group mean sd CV median IQR Ranger MAD ND 32.050 1.3625956 0.04251468 32.45 1.150 3.1 0.74130 HFD 37.875 0.8958236 0.02365211 37.70 0.825 2.1 0.66717 HFD_Extract_2X 33.350 3.3491292 0.10042367 33.35 5.750 5.9 4.29954 HFD_BC_Activated 33.750 3.7616486 0.11145625 32.35 3.450 8.1 1.63086 HFD_BC_Extract_1X 35.625 3.0966380 0.08692317 35.75 3.925 7.0 3.48411 HFD_BC_Extract_2X 30.000 1.6552945 0.05517648 30.10 1.450 4.0 1.55673

[0072] Figure 4 Table 4 shows the comparison of the levels of low-density lipoprotein cholesterol (LDL-C) in six groups of experimental mice after 18 weeks of intervention in the animal experiment of the present invention. Taking the mice in the high-fat control (HFD) group as the reference group, the levels of low-density lipoprotein cholesterol (LDL-C) in the group treated with 1X traditional Chinese medicine combined with bacterial liquid (HFD_BC_Extract_1X) and the group treated with 2X traditional Chinese medicine combined with bacterial liquid (HFD_BC_Extract_2X) were significantly reduced, but there were no significant differences between the 2X traditional Chinese medicine (HFD_Extract_2X) group and the high-fat control (HFD) group and the group treated with 2X traditional Chinese medicine combined with bacterial liquid (HFD_BC_Extract_2X), respectively.

[0073] Comparison of low-density lipoprotein cholesterol levels in mice of different groups 18 weeks after intervention

[0074] ND 0.3175 0.1161536 0.3658382 0.27 0.0775 0.25 0.029652 HFD 0.7175 0.1438460 0.2004822 0.72 0.2225 0.29 0.177912 HFD_Extract_2X 0.6975 0.3287730 0.4713591 0.67 0.4975 0.67 0.370650 HFD_BC_Activated 0.5825 0.1666083 0.2860229 0.56 0.2375 0.35 0.170499 HFD_BC_Extract_1X 0.4950 0.2180978 0.4406017 0.59 0.1450 0.46 0.044478 HFD_BC_Extract_2X 0.4800 0.1169045 0.2435511 0.48 0.1150 0.28 0.126021

[0075] Figure 5 Table 5 shows the comparison of subcutaneous fat levels in six groups of experimental mice provided for the animal experiment of the present invention 18 weeks after intervention. Taking the mice in the high-fat control (HFD) group as the reference group, the subcutaneous fat weight levels in the 2X traditional Chinese medicine (HFD_Extract_1X) group and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group were significantly reduced, but there was no significant difference between the 2X traditional Chinese medicine (HFD_Extract_2X) group and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group.

[0076] Table 5 Comparison of subcutaneous fat levels in mice of different groups 18 weeks after intervention

[0077] Group mean sd CV median IQR Ranger MAD ND 0.28625 0.13388895 0.4677343 0.2220 0.06975 0.273 0.0066717 HFD 1.02150 0.10850653 0.1062227 1.0445 0.10000 0.253 0.0778365 HFD_Extract_2X 0.60600 0.29797092 0.4917012 0.5630 0.42700 0.622 0.3009678 HFD_BC_Activated 0.63625 0.26392218 0.4148089 0.5600 0.28325 0.581 0.1704990 HFD_BC_Extract_1X 0.79275 0.32478955 0.4096998 0.6795 0.22425 0.726 0.1223145 HFD_BC_Extract_2X 0.29800 0.05109468 0.1714587 0.3190 0.03200 0.110 0.0118608

[0078] Figure 6 Table 6 shows the comparison of perirenal fat levels in six groups of experimental mice provided for the animal experiment of the present invention 18 weeks after intervention. Taking the mice in the high-fat control (HFD) group as the reference group, the perirenal fat weight level in the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group was significantly reduced, but there was no significant difference between the 2X traditional Chinese medicine (HFD_Extract_2X) group and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group.

[0079] Table 6 Comparison of perirenal fat levels in mice of different groups 18 weeks after intervention

[0080] Group mean sd CV median IQR Ranger MAD ND 0.12525 0.03782746 0.30201564 0.1315 0.03925 0.088 0.0318759 HFD 0.55725 0.05122093 0.09191732 0.5615 0.06675 0.114 0.0548562 HFD_Extract_2X 0.41400 0.17579723 0.42463099 0.3935 0.25700 0.365 0.1868076 HFD_BC_Activated 0.35250 0.05570458 0.15802717 0.3340 0.03300 0.126 0.0200151 HFD_BC_Extract_1X 0.52450 0.28253318 0.53867147 0.4250 0.17150 0.632 0.0934038

[0081]

[0082] Figure 7Table 7 shows the comparison of the epididymis fat levels of six groups of experimental mice provided for the animal experiments of the present invention 18 weeks after intervention. Taking the mice in the high-fat control (HFD) group as the reference group, the levels of epididymis fat weight in the 2X traditional Chinese medicine (HFD_Extract_2X) group and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group were significantly reduced, and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group was more significant (P<0.001), but there was no significant difference between the 2X traditional Chinese medicine (HFD_Extract_2X) group and the 2X traditional Chinese medicine combined with bacterial solution (HFD_BC_Extract_2X) group.

[0083] Table 7 Comparison of epididymis fat levels of mice in different groups 18 weeks after intervention

[0084] Group mean sd CV median IQR Ranger MAD ND 0.42750 0.1008382 0.23587872 0.4265 0.16650 0.189 0.1275036 HFD 2.10375 0.1854694 0.08816135 2.1220 0.22025 0.425 0.1868076 HFD_Extract_2X 1.34625 0.6098554 0.45300307 1.2610 0.89775 1.251 0.6263985 HFD_BC_Activated 1.35025 0.5506659 0.40782513 1.1310 0.43675 1.183 0.1831011 HFD_BC_Extract_1X 1.70500 0.5664362 0.33222066 1.6085 0.38550 1.359 0.3832521 HFD_BC_Extract_2X 0.65750 0.1673290 0.25449280 0.6315 0.15950 0.395 0.1378818

[0085] Body weight showed ( Figure 3 ), at the α = 0.1 test level, the body weight of the 2X traditional Chinese medicine combined with bacterial solution group was significantly lower than that of the simple 2X traditional Chinese medicine group (p = 0.073) and significantly lower than that of the simple bacterial solution group (p = 0.057); subcutaneous fat ( Figure 5 ) showed that the 2X traditional Chinese medicine combined with bacterial solution group was significantly lower than the 2X traditional Chinese medicine group (p = 0.014) and the bacterial solution group (p = 0.014); perirenal fat ( Figure 6 ) showed that the 2X traditional Chinese medicine combined with bacterial solution group was significantly lower than the 2X traditional Chinese medicine group (p = 0.021) and the bacterial solution group (p = 0.014); epididymis fat ( Figure 7 ) showed that the 2X traditional Chinese medicine combined with bacterial solution group was significantly lower than the bacterial solution group (p = 0.014) and significantly lower than the 2X traditional Chinese medicine group (p = 0.057) at the α = 0.1 test level. Comprehensive Figure 3 、 Figures 5 - 7 The index results show that the 2X traditional Chinese medicine combined with bacterial solution group has the best effect of weight loss and lipid reduction, and is better than the group using traditional Chinese medicine and bacterial solution alone, suggesting that the combination of bacterial solution and traditional Chinese medicine plays a synergistic effect in weight loss and lipid reduction.

[0086] From Figures 4 - 7 Combined, it can be seen that in multiple indicators such as low-density lipoprotein (LDL), subcutaneous fat weight, perirenal fat weight, and epididymis fat weight, the 2X traditional Chinese medicine combined with bacterial solution group is significantly lower than the high-fat control group, suggesting that the 2X traditional Chinese medicine combined with bacterial solution group plays an efficient role in lipid reduction.

[0087] The variance of the 2X traditional Chinese medicine group (HFD_Extract_2X) is relatively large, indicating that the effect of using the traditional Chinese medicine extract alone has high heterogeneity. However, in terms of multiple indicators reflecting body weight and fat weight, such as body weight, low-density lipoprotein (LDL), subcutaneous fat, perirenal fat, and epididymal fat, the variance of the bacterial liquid combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) is relatively small, showing the smallest data fluctuation and the best stability, suggesting that the combination of bacterial liquid and traditional Chinese medicine has a more stable weight loss and lipid-lowering effect.

[0088] Moreover, from the comparison of the above experimental data, among the results of multiple indicators such as body weight, subcutaneous fat, perirenal fat, and epididymal fat, there are significant differences between the bacterial liquid combined with 2X traditional Chinese medicine group and the high-fat control group and the bacterial liquid combined with 1X traditional Chinese medicine group respectively. However, there is no significant difference between the high-fat control group and the bacterial liquid combined with 1X traditional Chinese medicine group, indicating that the combination effect of the bacterial liquid and 2X traditional Chinese medicine is better.

[0089] Among the results of multiple indicators such as body weight, subcutaneous fat, perirenal fat, and epididymal fat, it is suggested that the weight loss and lipid-lowering effect of the bacterial liquid combined with 1X traditional Chinese medicine group is not significant, and it is even worse than that of the single bacterial liquid group and the traditional Chinese medicine group. Under the condition of fixed bacterial liquid concentration, the effect is significant after doubling the concentration of traditional Chinese medicine. Therefore, the concentration of traditional Chinese medicine should not be reduced. In addition, considering that excessive intake of mulberry leaves may cause discomfort such as nausea, abdominal pain, and diarrhea, it is not suitable to increase the dose too much.

[0090] Figure 8 The staining diagrams of liver cells and subcutaneous fat cells of the normal control group (ND) mice provided for the animal experiments of the present invention are as Figure 8 (upper) shown. The capsule structure of the liver tissue of the normal control group (ND) mice is clear and there is no obvious abnormality. There are relatively many hepatocytes with mild fatty degeneration in the parenchyma, with tiny round vacuoles in the cytoplasm. The hepatic sinusoids are not significantly dilated or compressed, and there is no obvious abnormality in the portal area between adjacent hepatic lobules. As Figure 8 (lower) shown. The white adipose tissue of the normal control group (ND) mice consists of unilocular adipocytes of different sizes aggregated together. The cells are filled with a large lipid droplet, the cell nucleus is flattened and round, and together with the cytoplasm, it is squeezed to one side of the cell by the lipid droplet, showing a crescent shape. The shape and size of the adipocytes are not significantly abnormal. The stroma contains loose connective tissue, abundant blood vessels and capillaries, with a very small amount of venous congestion, and no obvious inflammatory cell infiltration is seen.

[0091] Figure 9 The staining diagrams of liver cells and subcutaneous fat cells of the high-fat control group (HFD) mice provided for the animal experiments of the present invention are as Figure 9(Upper) As shown, the capsule structure of the liver tissue of the high-fat control group (HFD) mice was clear and there were no obvious abnormalities. A large number of hepatocytes in the parenchyma showed mild fatty degeneration, with round vacuoles of different sizes in the cytoplasm. There was a large amount of central vein and portal vein congestion. The hepatic sinusoids were not significantly dilated or compressed, and there were no obvious abnormalities in the portal areas between adjacent hepatic lobules. As Figure 9 (Lower) As shown, the white adipose tissue of the high-fat control group (HFD) mice consisted of unilocular adipocytes of different sizes aggregated together. The cells were filled with a large lipid droplet, the cell nucleus was flattened and round, and together with the cytoplasm, it was squeezed to one side of the cell by the lipid droplet, showing a crescent shape. A relatively large number of adipocytes had a relatively large volume. The stroma contained loose connective tissue, abundant blood vessels and capillaries, with no obvious abnormalities and no obvious inflammatory cell infiltration.

[0092] Figure 10 The stained sections of liver cells and subcutaneous adipocytes of the mice in the bacterial liquid combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) provided for the animal experiments of the present invention are as Figure 10 (Upper) As shown, the capsule structure of the liver tissue of the mice in the bacterial liquid combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) was clear and there were no obvious abnormalities. A small number of hepatocytes in the parenchyma showed slight fatty degeneration, with tiny round vacuoles in the cytoplasm. Occasional punctate necrosis of hepatocytes was seen, with nuclear fragmentation, accompanied by a very small amount of lymphocyte infiltration. The hepatic sinusoids were not significantly dilated or compressed, and there were no obvious abnormalities in the portal areas between adjacent hepatic lobules. As Figure 10 (Lower) As shown, the white adipose tissue of the mice in the bacterial liquid combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) consisted of unilocular adipocytes of different sizes aggregated together. The cells were filled with a large lipid droplet, the cell nucleus was flattened and round, and together with the cytoplasm, it was squeezed to one side of the cell by the lipid droplet, showing a crescent shape. There were no obvious abnormalities in the shape and size of the adipocytes; the stroma contained loose connective tissue, abundant blood vessels and capillaries.

[0093] Figures 8 - 10 The cell staining results showed that the state of the liver cells of the mice in the bacterial liquid combined with 2X traditional Chinese medicine group (HFD_BC_Extract_2X) was consistent with that of the normal control (ND) group, with less mild fatty degeneration compared to the high-fat control (HFD) group; compared with the size of the white adipocytes in the normal control (ND) group, the high-fat control (HFD) group became larger, while the bacterial liquid combined with 2X traditional Chinese medicine (HFD_BC_Extract_2X) group became smaller. This shows that the bacterial liquid combined with 2X traditional Chinese medicine group exerts the effects of weight loss and lipid reduction, and the adipocytes return to the normal control group level or even a lower level.

[0094] IV. Application Examples

[0095] First, adopt the process of multiple decoctions (purified water, filtered through 200-mesh sieve) + low-temperature rotary concentration at 65°C to 75°C to prepare a mulberry leaf / coix seed extract with a crude drug concentration of 0.24 - 0.48 g / ml (such as the dispensing mode of 1128 g of raw materials → 665 ml of concentrated solution), that is, a traditional Chinese medicine extract;

[0096] Then, mix the Blautia coccoides bacterial solution (1×10 8 CFU / mL) with the traditional Chinese medicine extract at 2X concentration in a volume ratio of 1:3, and anaerobically culture at about 36°C for about 72 hours (referring to the ratio of 5 ml of bacterial solution + 15 ml of culture medium in the in vitro interaction test) to obtain a product of combined bacteria and traditional Chinese medicine.

[0097] Develop this product into a probiotic preparation or a product for intervening in metabolic diseases; or, based on the optimization of the concentration gradient (2X / 1X / 0.5X / 0.1X) and the ratio of the bacterial solution, develop customized compositions for different BMI classifications, and achieve personalized medicine through precise nutritional intervention.

[0098] Alternatively, the freeze-drying can be carried out on the dispensed traditional Chinese medicine extract and the bacterial agent mixture to prepare an instant solid beverage; or, develop the above product into a weight management food, such as a meal replacement powder or a functional beverage, with each portion containing an equivalent of 0.24 g / ml of extract + 1×10 8 CFU / mL of bacterial concentration. For the functional beverage, each portion is in the range of 200 - 400 ml, and for the meal replacement powder, each portion is in the range of 30 - 50 g.

[0099] The above are only some embodiments of the present invention. For those of ordinary skill in the art, without departing from the inventive concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. Application of Blautia coccoides in combination with traditional Chinese medicine extract in the preparation of drugs or foods, characterized in that, The drug or food has any of the following functions: (a) Weight loss; (b) Lipid reduction; The traditional Chinese medicine extract is mulberry leaf extract and coix seed extract. The preservation number of the Blautia coccoides is ATCC29236, and the name is Blautia coccoides.

2. The application according to claim 1, wherein The mulberry leaf extract is prepared by the following method: (1) Put mulberry leaves into water, decoct and filter to obtain the first filter residue and the first filtrate; (2) Add water to the first filter residue, decoct again and filter to obtain the second filtrate; (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain.

3. The application according to claim 1, characterized in that, The coix seed extract is prepared by the following method: (1) Put the soaked coix seeds into water, decoct and then filter to obtain the first filter residue and the first filtrate; (2) Add water to the first filter residue, decoct again and filter to obtain the second filtrate; (3) Combine the two filtrates and concentrate to a crude drug concentration of 0.36 g / mL to 0.5 g / mL to obtain.

4. The application according to any one of claims 1 to 3, characterized in that, The Blautia coccoides needs to be activated by the following steps before use: Inoculate Blautia coccoides into the improved chopped meat medium of ATCC 1490, and after culturing for 48 to 72 hours under anaerobic conditions at 36°C to 37°C, detect the bacterial liquid concentration, and adjust the bacterial liquid concentration to 1×10 8 CFU / mL to obtain the activated Blautia coccoides bacterial liquid.

5. The application according to claim 4, characterized in that, The volume ratio of the Blautia coccoides liquid to the traditional Chinese medicine extract in the drug or food is 1:2 to 4.

6. The application according to claim 5, wherein The lipid reduction includes reducing at least one of low-density lipoprotein, subcutaneous fat, perirenal fat, and epididymal fat.

7. A weight loss and lipid-lowering drug, characterized in that, Its active ingredients are Blautia coccoides bacterial liquid and traditional Chinese medicine extract, and the volume ratio of the two is 1:3; the traditional Chinese medicine extract is mulberry leaf extract and coix seed extract, and the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.48 g / mL to 0.5 g / mL; the concentration of the Blautia coccoides bacterial liquid is 1×10 8 CFU / mL.

8. A functional food for weight loss and lipid reduction, characterized in that, Its active ingredients are Blautia coccoides bacterial liquid and traditional Chinese medicine extract, and the volume ratio of the two is 1:3; the traditional Chinese medicine extract is mulberry leaf extract and coix seed extract, and the crude drug concentrations of the mulberry leaf extract and coix seed extract are both 0.48 g / mL to 0.5 g / mL; the concentration of the Blautia coccoides bacterial liquid is 1×10 8 CFU / mL.

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