Probiotic Chinese herbal medicine compound composition as well as preparation method and application thereof

Abstract

The invention discloses a probiotic Chinese herbal medicine compound composition which comprises a Chinese herbal medicine compound and probiotics, wherein the Chinese herbal medicine compound comprises a lotus leaf extract, a hawthorn extract and a broadleaf holly leaf extract; and the probiotics are bifidobacteria. The invention further discloses a preparation method of the probiotic Chinese herbal medicine compound composition and application of the probiotic Chinese herbal medicine compound composition in preparation of fat-reducing products. The probiotic Chinese herbal medicine compound composition provided by the invention not only has a remarkable improvement effect on fat accumulation, blood fat and liver metabolism disorder caused by high-fat diet, but also has a remarkable regulation effect on liver sterol metabolism, sterol biosynthesis related genes and steroid biosynthesis pathways.

Description

Technical Field

[0001] The present invention relates to the technical field of health food, and in particular to a fat-reducing probiotic Chinese herbal medicine compound composition, a preparation method and application thereof. Background Art

[0002] With the change of modern lifestyle and dietary structure, obesity and its accompanying metabolic disorders are becoming more and more common around the world and have become an important public health issue that seriously threatens human health. Obesity is not only manifested as weight gain, but also accompanied by abnormal accumulation of visceral fat, significant increase in blood lipid levels and disorder of liver lipid metabolism, which in turn induces metabolic diseases such as hypertension, hyperglycemia and hyperlipidemia, and is even closely related to the occurrence of chronic diseases such as diabetes and atherosclerosis. These diseases can significantly increase the mortality rate and life burden of patients, so controlling obesity and its related metabolic diseases has become a research focus of clinical medicine and nutritional science.

[0003] Probiotics have been widely used to improve lipid metabolism because of their role in regulating intestinal flora and promoting metabolic health. Studies have shown that specific probiotics can alleviate obesity-related symptoms to a certain extent by affecting cholesterol metabolism and lipid absorption. However, the effectiveness of probiotics depends on their survival rate and activity in the intestine. A sufficient number of active probiotics must be ensured to achieve the desired metabolic regulation effect. Generally, the number of active probiotics in functional foods should not be less than 10 6 CFU / mL to ensure that it passes smoothly through the gastrointestinal tract and exerts its effect in the intestine. However, since the number and activity of probiotics are easily affected during product preparation and storage, single probiotic products often face limitations in achieving fat-reducing effects.

[0004] Chinese herbal compound conditioning has occupied an important position in traditional Chinese medicine since ancient times, especially in blood lipid regulation and body fat management. Modern pharmacological studies have confirmed that a variety of Chinese herbal ingredients can have a positive impact on lipid metabolism through multi-target mechanisms, thereby exerting lipid-lowering, anti-oxidation, anti-inflammatory and other effects. By compounding probiotics with Chinese herbal medicines to form a synergistic composition, it is possible to comprehensively regulate intestinal flora and host lipid metabolism. This method has great application potential and market value. In particular, Chinese herbal medicines with lipid-lowering potential such as lotus leaves, hawthorn and kudingcha, their combination with specific probiotics may provide an efficient solution for fat reduction and metabolic regulation.

[0005] However, there are still many technical problems to be solved in the development of Chinese herbal probiotic compositions for fat loss. For example, the optimal combination of probiotics and Chinese herbal medicines suitable for efficient fat loss, the reasonable dosage ratio, the synergistic mechanism of the active ingredients, etc. are still unclear. At the same time, how to ensure the stability and efficacy of active ingredients in product preparation and application is still a major challenge in research and development. Therefore, the development of a probiotic Chinese herbal compound composition with a reasonable formula, significant efficacy and clear mechanism, which can provide a scientific solution to metabolic disorders and fat accumulation caused by a high-fat diet, is a problem that technicians in the current field need to overcome urgently. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a probiotic Chinese herbal medicine compound composition, which not only has a significant improvement effect on fat accumulation, blood lipids and liver metabolic disorders caused by a high-fat diet, but also has a significant regulatory effect on liver steroid metabolism and steroid biosynthesis-related genes and steroid biosynthesis pathways.

[0007] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0008] The first aspect of the present invention provides a probiotic Chinese herbal compound composition, comprising a Chinese herbal compound and probiotics; wherein the Chinese herbal compound comprises lotus leaf extract, hawthorn extract and kudingcha extract; and the probiotics are bifidobacteria.

[0009] Lotus leaves contain various alkaloids such as nelumboine, protonemucoside, nelumboine and flavonoids, as well as vitamin C, which have the effects of clearing heat, diuresis, clearing fat and relieving constipation.

[0010] Kuding tea contains kuding saponins, amino acids, vitamin C, polyphenols, flavonoids, caffeine, protein and other ingredients. It has many functions such as clearing away heat and relieving summer heat, diuresis and strengthening the heart, moistening the throat and relieving cough, lowering blood pressure and losing weight, and activating blood circulation.

[0011] Hawthorn contains flavonoids, citric acid, malic acid, ascorbic acid and carbohydrates, which can lower blood pressure and promote gastrointestinal fat excretion. Traditional Chinese medicine believes that hawthorn can strengthen the spleen and eliminate stagnation, is beneficial for weight loss, and can assist in the treatment of obesity.

[0012] Bifidobacterium can promote fat metabolism and reduce fat accumulation through various pathways. It can improve the balance of intestinal microecology, inhibit the growth of harmful bacteria, reduce intestinal inflammation, and thus promote fat metabolism; by improving insulin sensitivity, regulating insulin receptors and insulin signaling pathways, it helps to control weight and reduce fat accumulation; by regulating fatty acid metabolism, it promotes the oxidation of short-chain fatty acids (such as acetate, propionate, butyrate), inhibiting the accumulation of fat cells; in addition, bifidobacteria can also reduce chronic low-grade inflammation of adipose tissue, reduce fat synthesis and adipocyte differentiation, regulate intestinal barrier function, enhance fatty acid oxidation and improve blood sugar levels. These effects work together on fat metabolism to exert a fat-reducing effect, and the main targets include PPARs, AMPK, SCFA receptors and genes related to fatty acid synthesis.

[0013] The inventors found that when the Chinese herbal medicine complex - lotus leaf extract, hawthorn extract and kudingcha extract, is used in combination with bifidobacteria, their fat-reducing efficacy can be significantly enhanced. It not only has a significant improvement effect on fat accumulation, blood lipids and liver metabolic disorders caused by a high-fat diet, but also has a significant regulatory effect on liver steroid metabolism and steroid biosynthesis-related genes and steroid biosynthesis pathways.

[0014] Specifically, the probiotic Chinese herbal medicine compound composition of the present invention significantly reduces the weight of epididymal fat, mesenteric fat and inguinal fat, significantly improves the reduction of serum high-density lipoprotein cholesterol (HDLC) levels and the reduction of glutathione peroxidase (GPX) activity caused by a high-fat diet, and can significantly reduce triglyceride (TG) content and total cholesterol (TC) content in the liver. The results of liver morphological observation show that the probiotic Chinese herbal medicine compound composition of the present invention significantly reduces the number and size of lipid droplets in the liver, has a certain improvement effect on excessive liver fat deposition caused by a high-fat diet; and significantly reduces the volume of epididymal adipocytes.

[0015] Furthermore, the Chinese herbal medicine compound comprises the following components by mass percentage: 1-10% lotus leaf extract, 80-98.7% hawthorn extract, and 0.1-5% kudingcha extract. Preferably, the mass ratio of lotus leaf extract, hawthorn extract, and kudingcha extract in the Chinese herbal medicine composition is 3:24:2.

[0016] In the present invention, the probiotics may be one or more of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum and Bifidobacterium breve.

[0017] In the present invention, the probiotics in the composition are bifidobacterium powder. Further, the bifidobacterium powder is self-prepared bifidobacterium freeze-dried powder, and its bacterial content is 1×10 10The present invention does not specifically limit the source of probiotics, and one or more bifidobacterium strains commonly found on the market or self-prepared bifidobacterium strains can be used according to demand, and the source of the strain does not affect the implementation effect of the present invention.

[0018] Furthermore, in the probiotic Chinese herbal medicine compound composition, the mass ratio of the Chinese herbal medicine compound to the probiotics is: 72-90% of the Chinese herbal medicine compound and 10-28% of the probiotics. For example, in the probiotic Chinese herbal medicine compound composition, the mass percentage of the Chinese herbal medicine compound can be 72%, 75%, 78%, 80%, 82%, 85%, 88%, 90%, etc., and the mass percentage of the probiotics can be 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28%, etc.

[0019] The second aspect of the present invention provides a method for preparing the probiotic Chinese herbal medicine compound composition, comprising the following steps:

[0020] S1. The extract raw material is extracted twice with water, and impurities are filtered out after each extraction to obtain an extract; the extract raw material is lotus leaf powder, hawthorn powder or kudingcha powder;

[0021] S2. The extract is concentrated under vacuum to a relative density of 1.09 to 1.12 g / cm 3 , then add maltodextrin and sterilize by high pressure;

[0022] S3. In a clean environment, the sterilized extract is spray-dried to obtain a powdered material; the powdered material is crushed, sieved, and mixed to obtain an extract powder;

[0023] S4. Mixing the extract powder with probiotic powder to obtain the probiotic Chinese herbal medicine compound composition.

[0024] Furthermore, in step S1, when the extract raw material is lotus leaf powder, the lotus leaf powder is soaked in 30 times and 20 times water, respectively, and the extraction time is 120 minutes and 60 minutes, respectively;

[0025] When the extract raw material is hawthorn powder, 12 times and 8 times of water are used to soak the hawthorn powder, and the extraction time is 120 minutes and 60 minutes respectively;

[0026] When the extract raw material is Kudingcha powder, 12 times and 8 times of water are used to soak the Kudingcha powder, and the extraction time is 90 minutes and 60 minutes, respectively.

[0027] Furthermore, in step S2, the vacuum degree is controlled to be -0.06 to -0.09 MPa during the concentration, and the concentration temperature is 50-75°C; and the temperature is controlled to be 115-135°C during the high-pressure sterilization.

[0028] Furthermore, in step S3, during spray drying, the air inlet temperature is 180-205°C, and the air outlet temperature is 85-105°C.

[0029] In some embodiments of the present invention, taking lotus leaf extract as an example, the specific extraction process is as follows:

[0030] a1. Raw material source and testing: The lotus leaf powder raw material is confirmed by QC testing and prepared for the water extraction process.

[0031] a2. Extraction and filtration: Use water extraction twice, soak the lotus leaf powder in 30 times and 20 times water respectively, the extraction time is 120 minutes and 60 minutes, filter after each extraction to remove insoluble impurities.

[0032] a3. Concentration and sterilization: The filtered extract is concentrated under vacuum conditions, with the vacuum degree controlled at -0.06 to -0.09 MPa and the concentration temperature at 50-75°C. The relative density of the final concentrate is 1.09-1.12. Maltodextrin is added to the concentrate, which is sterilized by high pressure at a temperature of 115-135°C.

[0033] a4. Drying and crushing: The sterilized extract is spray dried in a clean production area (Class D), with an inlet air temperature of 180-205°C and an outlet air temperature of 85-105°C. After drying, a powdery material is obtained, which is crushed and passed through a 100-mesh sieve.

[0034] a5. Screening, mixing and testing: Screen again through a 100-mesh sieve under CCP standard control and mix for 50-60 minutes. The lotus leaf powder extract is tested internally, sealed for inner packaging, and then packaged externally after being confirmed qualified by CCP inspection and sent to the warehouse for storage.

[0035] The third aspect of the present invention provides the use of the above-mentioned probiotic Chinese herbal medicine compound composition in the preparation of fat-reducing products.

[0036] Furthermore, the fat-reducing product is used to improve fat accumulation, blood lipids and liver metabolic disorders caused by a high-fat diet.

[0037] Furthermore, the fat-reducing product is used to regulate liver steroid metabolism and steroid biosynthesis-related genes and steroid biosynthesis pathways.

[0038] Further, the dosage of the probiotic Chinese herbal compound composition is: the dosage of the Chinese herbal compound is 10-24 g / d, for example, 10 g / d, 12 g / d, 14 g / d, 16 g / d, 18 g / d, 20 g / d, 22 g / d, 24 g / d, etc. The dosage of the probiotic is 1-6 g / d, for example, 1 g / d, 2 g / d, 3 g / d, 4 g / d, 5 g / d, 6 g / d, etc.

[0039] When determining the dosage of probiotics, the final dosage can be adjusted within this range according to individual weight and required probiotic intake to meet specific health needs. Preferably, the dosage of probiotics is 2 g / d.

[0040] Furthermore, the fat-reducing products include but are not limited to fat-reducing medicines, fat-reducing foods and fat-reducing health products.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1. The probiotic Chinese herbal medicine compound composition provided by the present invention has a significant improvement effect on fat accumulation, blood lipids and liver metabolic disorders caused by a high-fat diet. The probiotic Chinese herbal medicine compound composition significantly reduces the weight of epididymal fat, mesenteric fat and inguinal fat, significantly improves the reduction of serum high-density lipoprotein cholesterol (HDLC) levels and the reduction of glutathione peroxidase (GPX) activity caused by a high-fat diet, and can also significantly reduce triglyceride (TG) content and total cholesterol (TC) content in the liver. In addition, morphological observation results show that the probiotic Chinese herbal medicine compound composition significantly reduces the number and size of fat droplets in the liver and significantly reduces the volume of epididymal adipocytes.

[0043] 2. The probiotic Chinese herbal medicine compound composition provided by the present invention has a significant regulatory effect on liver steroid metabolism, steroid biosynthesis-related genes and steroid biosynthesis pathways.

[0044] 3. The probiotic Chinese herbal medicine compound composition provided by the present invention can be used to prepare various fat-reducing foods and fat-reducing health products. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 The bar graph is a graph showing the effect of the probiotic Chinese herbal medicine compound composition on epididymal fat weight in mice;

[0046] Figure 2 It is a bar graph showing the effect of the probiotic Chinese herbal compound composition on the mesenteric fat weight of mice;

[0047] Figure 3 It is a bar graph showing the effect of the probiotic Chinese herbal compound composition on the inguinal fat weight of mice;

[0048] Figure 4 It is a bar graph showing the effect of the probiotic Chinese herbal medicine compound composition on the serum high-density lipoprotein cholesterol (HDLC) level of mice;

[0049] Figure 5 It is a bar graph showing the effect of the probiotic Chinese herbal compound composition on the triglyceride (TG) content in the liver;

[0050] Figure 6 A bar graph showing the effect of the probiotic Chinese herbal medicine compound composition on the total cholesterol (TC) content in the liver;

[0051] Figure 7 It is a bar graph showing the effect of the probiotic Chinese herbal compound composition on the activity of glutathione peroxidase (GPX) in the liver;

[0052] Figure 8 HE staining photos of the effects of the probiotic Chinese herbal medicine compound composition on liver cell morphology;

[0053] Fig. 9 HE staining photos of the effects of the probiotic Chinese herbal medicine compound on the morphology of epididymal adipocytes;

[0054] Fig.10 The volcano map of the differentially expressed genes in the liver of the probiotic Chinese herbal medicine compound composition;

[0055] Fig.11 This is the bar graph of GO enrichment analysis of the probiotic Chinese herbal medicine compound composition on the differentially expressed genes in the liver;

[0056] Fig.12 This is a scatter plot of KEGG enrichment analysis of the probiotic Chinese herbal compound composition on differentially expressed genes in the liver. DETAILED DESCRIPTION

[0057] The technical solutions in the embodiments of the present invention will be described clearly and completely below in combination with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0058] Example 1

[0059] This embodiment provides a probiotic Chinese herbal medicine compound composition, including a Chinese herbal medicine compound (TCM compound) and probiotics;

[0060] The Chinese herbal medicine compound includes lotus leaf extract, hawthorn extract and kudingcha extract, and their mass ratio is 3:24:2. The probiotics are freeze-dried powder of Bifidobacterium bifidum, and the bacterial content is 1*10 10 CFU / g.

[0061] Comparative Example 1

[0062] This comparative example provides a Chinese herbal medicine complex (TCM compound), including lotus leaf extract, hawthorn extract and Kudingcha tea extract, and their mass ratio is 3:24:2.

[0063] Comparative Example 2

[0064] This comparative example provides a probiotic freeze-dried powder, which is a freeze-dried powder of Bifidobacterium bifidum, with a bacterial content of 1×10 10 CFU / g.

[0065] Animal feeding experiments

[0066] 6-week-old male C57BL / 6 mice were raised in an SPF rodent breeding room. The temperature of the breeding environment was controlled at 22±2℃, with a 12-hour light-dark cycle and an environmental relative humidity of 40-60%. After 1 week (7 days) of adaptive feeding, the experimental mice were randomly divided into 7 experimental groups (7 groups in total, 10 mice in each group). Each group was fed with a different diet. During the experiment, the mice were free to eat and drink water for 8 weeks (56 days). According to the body surface area coefficient conversion: the dose of the mouse (0.020kg) animal experiment was converted to the human (60.0kg) dose of about 1 / 268.1, so the total dose of the experimental group was controlled to 10mg / d / pcs.

[0067] The specific experimental group design is:

[0068] (1) Normal diet group, fed with normal mouse basal feed with a fat energy supply ratio of 10%, denoted as Chow;

[0069] (2) high-fat diet group, fed with purified high-fat feed with a fat energy supply ratio of 60%, recorded as HFD;

[0070] (3) High-fat diet + probiotics group, fed with purified high-fat feed (fat energy supply ratio 60%), and intragastrically administered with a quantitative amount of probiotic Bifidobacterium lyophilized powder of comparative example 2 (at a dosage of 10 mg / d / pcs) every day, recorded as HFD + probiotics;

[0071] (4) High-fat diet + TCM compound group, fed with purified high-fat feed added with TCM compound of comparative example 1 (fat energy ratio 60%, TCM compound addition amount 0.38g / 100g - equivalent to 10mg / d / pcs per head), recorded as HFD+TCM;

[0072] (5) High-fat diet + TCM compound + probiotics group: The rats were fed with purified high-fat feed supplemented with the TCM compound of Example 1 (fat energy supply ratio 60%, TCM compound addition amount of 0.33 g / 100 g, equivalent to 8.8 mg / d / pcs per rat); the rats were gavaged with a fixed amount of probiotics of Example 1 every day (gavage at a dosage of 1.2 mg / d / pcs), recorded as HFD+TCM+probiotics.

[0073] Adipose tissue assessment procedure

[0074] To assess the weight of fat reserves (including epididymal, mesenteric, and inguinal fat) of each group of mice, the following procedures were performed:

[0075] (1) Experimental preparation: Sterilize and calibrate all dissection tools, and label the collection tubes for adipose tissue (EP tubes).

[0076] (2) Euthanasia and dissection: Euthanasia was performed in accordance with the ethical standards for experimental animals. The mice were placed in the supine position and the abdomen was incised along the midline.

[0077] (3) Separation of epididymal adipose tissue: Gently pull back the intestine to locate and separate the fat around the epididymis.

[0078] (4) Separation of mesenteric fat tissue: Find the fat surrounding the mesentery and carefully remove it.

[0079] (5) Inguinal adipose tissue separation: Enter the groin from under the skin and obtain the fat in the groin area.

[0080] (6) Weighing and recording: Transfer each adipose tissue into a pre-weighed tube, record the weight using a high-precision balance, and repeat the weighing to obtain the exact weight of each mouse adipose tissue.

[0081] (7) This experiment systematically analyzed the effects of each treatment group on mouse fat metabolism through precise fat weighing data to evaluate the efficacy and mechanism of the combined use of Chinese herbal compound and probiotics.

[0082] 1. Effects of probiotic Chinese herbal medicine on epididymal fat, mesenteric fat, and inguinal fat weight

[0083] Figure 1 The bar graph shows the effect of the probiotic Chinese herbal compound on epididymal fat weight in mice. Figure 1 It can be seen that when fed with a normal diet, the epididymal fat weight of mice was low. When fed with a high-fat diet, the epididymal fat weight of mice increased significantly. When the Chinese herbal compound TCM was added to the high-fat diet, or when the probiotic Bifidobacterium was gavaged daily, the epididymal fat weight of mice was significantly reduced, indicating that both Bifidobacterium and the Chinese herbal compound TCM are beneficial to reducing the epididymal fat weight of mice.

[0084] Compared with the other groups, the HFD+TCM+probiotics group, which added the Chinese herbal compound TCM to the high-fat diet and gavaged the probiotic Bifidobacterium daily, had the most significant effect in reducing the epididymal fat weight of mice. Specifically, compared with the HFD group, the epididymal fat weight of mice decreased from 31.47 mg / g BW to 17.64 mg / g BW, a reduction of 43.94%.

[0085] Figure 2 The bar graph shows the effect of the probiotic Chinese herbal medicine compound on the mesenteric fat weight of mice. Figure 2 It can be seen that when fed with a normal diet, the mesenteric fat weight of mice was low. When fed with a high-fat diet, the mesenteric fat weight of mice increased significantly. When the Chinese herbal compound TCM was added to the high-fat diet, or when the probiotic Bifidobacterium was gavaged daily, the mesenteric fat weight of mice was significantly reduced, indicating that both Bifidobacterium and the Chinese herbal compound TCM are beneficial to reducing the mesenteric fat weight of mice.

[0086] Compared with the other groups, the HFD+TCM+probiotics group, which added the Chinese herbal compound TCM to the high-fat diet and gavaged the probiotic Bifidobacterium daily, had the most significant effect in reducing the mesenteric fat weight of mice. Specifically, compared with the HFD group, the mesenteric fat weight of mice decreased from 8.24 mg / g BW to 4.25 mg / g BW, a reduction of 48.42%.

[0087] Figure 3 The bar graph shows the effect of probiotic Chinese herbal compound on inguinal fat weight in mice. Figure 3 It can be seen that when fed with a normal diet, the inguinal fat weight of mice was low. When fed with a high-fat diet, the inguinal fat weight of mice increased significantly. When the Chinese herbal compound TCM was added to the high-fat diet, or when the probiotic Bifidobacterium was gavaged daily, the inguinal fat weight of mice was significantly reduced, indicating that both Bifidobacterium and the Chinese herbal compound TCM are beneficial for reducing the inguinal fat weight of mice.

[0088] Compared with the other groups, the HFD+TCM+probiotics group, which added the Chinese herbal compound TCM to the high-fat diet and gavaged the probiotic Bifidobacterium daily, had the most significant effect in reducing the inguinal fat weight of mice. Specifically, compared with the HFD group, the inguinal fat weight of mice decreased from 26.17 mg / g BW to 11.00 mg / g BW, with a reduction percentage of 57.99%.

[0089] 2. Effect of probiotic Chinese herbal medicine on serum high-density lipoprotein cholesterol (HDLC) levels

[0090] Figure 4 This is a bar graph showing the effect of the probiotic Chinese herbal compound on the serum high-density lipoprotein cholesterol (HDLC) level of mice. A decrease in HDLC level is an important feature of dyslipidemia. As can be seen from the figure, when fed with a normal diet, the HDLC level of mice was about 2.9mmol / L; when fed with a high-fat diet, the HDLC of mice decreased significantly. Adding the Chinese herbal compound TCM to the high-fat diet, or gavage with the probiotic Bifidobacterium every day, the HDLC of mice increased significantly, indicating that both Bifidobacterium and the Chinese herbal compound TCM are beneficial to increase the HDLC of mice.

[0091] Compared with the other groups, the HFD+TCM+probiotics group, which added the Chinese herbal compound TCM to the high-fat diet and gavaged the probiotic Bifidobacterium every day, had the most significant effect in increasing the HDLC of mice. Specifically, compared with the HFD group, the HDLC of mice increased from 1.67mmol / L to 4.12mmol / L, with an increase of 146.71%.

[0092] 3. Effect of probiotic Chinese herbal medicine on triglyceride (TG) content in liver

[0093] Figure 5 The bar graph shows the effect of the probiotic Chinese herbal compound on triglyceride (TG) content in the liver. Figure 5 It can be seen that when fed with a normal diet, the triglyceride content in the mouse liver is low. When fed with a high-fat diet, the triglyceride content in the mouse liver increased significantly. Adding the Chinese herbal compound TCM to the high-fat diet or gavaging the probiotic Bifidobacterium daily did not reduce the triglyceride content in the mouse liver, which shows that the use of Bifidobacterium or the Chinese herbal compound TCM alone will not reduce the triglyceride content in the mouse liver.

[0094] Compared with the other groups, the content of triglycerides in the liver of mice in the HFD+TCM+probiotics group was significantly reduced by adding the Chinese herbal compound TCM to the high-fat feed and gavaging the probiotic bifidobacterium every day. Specifically, compared with the HFD group, the triglyceride (TG) content in the mouse liver decreased from 0.86mmol / L to 0.50mmol / L, and the percentage of reduction reached 41.7%. This shows that when bifidobacteria and the Chinese herbal compound TCM are used in combination, the content of triglycerides in the mouse liver can be significantly reduced.

[0095] 4. Effect of probiotic Chinese herbal medicine on total cholesterol (TC) content in liver

[0096] Figure 6 The bar graph shows the effect of the probiotic Chinese herbal compound on the total cholesterol (TC) content in the liver. Figure 6 It can be seen that when fed with a normal diet, the total cholesterol content in the mouse liver is low. When fed with a high-fat diet, the total cholesterol content in the mouse liver increased significantly. When the probiotic Bifidobacterium was gavaged daily while feeding a high-fat diet, the total cholesterol content in the mouse liver decreased to a certain extent. When the Chinese herbal compound TCM was added to the high-fat diet, the total cholesterol content in the mouse liver decreased significantly. This shows that compared with probiotics, the Chinese herbal compound TCM can better reduce the total cholesterol content in the mouse liver.

[0097] Compared with the other groups, the total cholesterol content in the liver of mice in the HFD+TCM+probiotics group was most significantly reduced in the group that added the Chinese herbal compound TCM to the high-fat diet and gavaged the probiotic Bifidobacterium every day. Specifically, compared with the HFD group, the total cholesterol content in the liver of mice decreased from 0.15mmol / L to 0.07mmol / L, a decrease of 54.2%.

[0098] 5. Effect of probiotic Chinese herbal medicine on glutathione peroxidase (GPX) activity in liver

[0099] Figure 7 The bar graph shows the effect of probiotic Chinese herbal compound on glutathione peroxidase (GPX) activity in the liver. Figure 7 It can be seen that when fed with a normal diet, the content of GPX in the mouse liver is high. When fed with a high-fat diet, the content of GPX in the mouse liver decreased. When the probiotic Bifidobacterium was gavaged daily while feeding a high-fat diet, the content of GPX in the mouse liver increased to a certain extent. When the Chinese herbal compound TCM was added to the high-fat diet, the content of GPX in the mouse liver increased significantly. This shows that compared with probiotics, the Chinese herbal compound TCM can better increase the content of GPX in the mouse liver.

[0100] Compared with the other groups, the increase in GPX content in the liver of mice in the HFD+TCM+probiotics group was the most significant. Specifically, compared with the HFD group, the GPX content in the liver of mice increased from 5.43mmol / L to 28.33mmol / L, an increase of 421.73%.

[0101] 6. Effects of probiotic Chinese herbal medicine on mouse liver cell morphology

[0102] Figure 8HE staining photos of the effects of probiotics and Chinese herbal medicine compound on liver cell morphology. The HE staining results in the figure show that the liver cells of mice in the Chow group are tightly arranged, while the livers of mice in the HFD group have obvious cavitation. The white vacuoles in the sections are the lipid droplets deposited in the liver. The probiotics and TCM compound groups significantly reduced the number and size of lipid droplets, and had a certain improvement effect on the excessive liver fat deposition caused by a high-fat diet. Among them, the high-fat diet + TCM compound + probiotics group had the most significant improvement effect.

[0103] 7. Effect of probiotic Chinese herbal medicine on epididymal adipocyte morphology in mice

[0104] Fig. 9 HE staining photos of the effects of probiotic Chinese herbal medicine compound on epididymal adipocyte morphology. The HE staining results in the figure show that the epididymal adipocytes in the Chow group were small in size, clear in morphology, and complete in structure; in contrast, the epididymal adipocytes in the HFD group were significantly increased. The addition of probiotics and TCM compound significantly reduced the volume of epididymal adipocytes, and the number of cells observed under the microscope field of view of the same magnification increased significantly. Among them, the high-fat diet + TCM compound + probiotic group had the most significant effect on improving epididymal adipocyte hypertrophy.

[0105] 8. Effects of probiotic Chinese herbal medicine on gene expression in mouse liver

[0106] Fig.10 This is a volcano map of the differentially expressed genes in the liver of probiotics and Chinese herbal medicines. The results of the transcriptomic analysis of mouse liver in the figure showed that compared with the HFD group, the high-fat diet + TCM compound + probiotics group had a total of 647 differentially expressed genes, including 324 up-regulated genes and 323 down-regulated genes.

[0107] Fig.11 The results of GO enrichment analysis showed that the differentially expressed genes were concentrated in the secondary alcohol biosynthesis process, cholesterol biosynthesis process, and sterol biosynthesis process.

[0108] Fig.12 The results of KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in pathways related to the biosynthesis of terpenoid skeletons, circadian rhythm, bile secretion and retinol metabolism.

[0109] In summary, the probiotic Chinese herbal medicine compound composition provided by the present invention not only has a significant improvement effect on fat accumulation, blood lipids and liver metabolic disorders caused by a high-fat diet, but also has a significant regulatory effect on liver steroid metabolism and steroid biosynthesis-related genes and steroid biosynthesis pathways, and can be used to prepare various fat-reducing medicines, fat-reducing foods, and fat-reducing health products.

[0110] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A probiotic Chinese herbal medicine compound composition, characterized in that: Includes Chinese herbal complexes and probiotics; The Chinese herbal medicine compound comprises lotus leaf extract, hawthorn extract and kudingcha extract; The probiotics are bifidobacteria.

2. A probiotic Chinese herbal medicine compound composition according to claim 1, characterized in that: The Chinese herbal medicine compound comprises the following components by mass percentage: 1-10% lotus leaf extract, 80-98.7% hawthorn extract and 0.1-5% kudingcha tea extract.

3. A probiotic Chinese herbal medicine compound composition according to claim 2, characterized in that: In the Chinese herbal medicine complex, the mass ratio of lotus leaf extract, hawthorn extract and Kudingcha tea extract is 3:24:

2.

4. The probiotic Chinese herbal medicine compound composition according to claim 1, characterized in that: The probiotics include one or more of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum and Bifidobacterium breve; or, The probiotics are bifidobacterium powder, and the bacterial content is 1*10 10 CFU / g.

5. The probiotic Chinese herbal medicine compound composition according to claim 1, characterized in that: In the probiotic Chinese herbal medicine compound composition, the mass percentages of the Chinese herbal medicine compound and the probiotics are respectively: 72-90% of the Chinese herbal medicine compound and 10-28% of the probiotics.

6. The method for preparing the probiotic Chinese herbal medicine compound composition according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1. The extract raw material is extracted twice with water, and impurities are filtered out after each extraction to obtain an extract; the extract raw material is lotus leaf powder, hawthorn powder or kudingcha powder; S2. The extract is concentrated under vacuum to a relative density of 1.09 to 1.12 g / cm 3 , then add maltodextrin and sterilize by high pressure; S3. In a clean environment, the sterilized extract is spray-dried to obtain a powdered material; the powdered material is crushed, sieved, and mixed to obtain an extract powder; S4. Mixing the extract powder with probiotic powder to obtain the probiotic Chinese herbal medicine compound composition.

7. The method for preparing the probiotic Chinese herbal medicine compound composition according to claim 6, characterized in that: In step S1, when the extract raw material is lotus leaf powder, the lotus leaf powder is soaked in 30 times and 20 times water, respectively, and the extraction time is 120 minutes and 60 minutes, respectively; When the extract raw material is hawthorn powder, 12 times and 8 times of water are used to soak the hawthorn powder, and the extraction time is 120 minutes and 60 minutes respectively; When the extract raw material is Kudingcha powder, 12 times and 8 times of water are used to soak the Kudingcha powder, and the extraction time is 90 minutes and 60 minutes, respectively.

8. The method for preparing the probiotic Chinese herbal medicine compound composition according to claim 6, characterized in that: In step S2, the vacuum degree is controlled to be -0.06 to -0.09 MPa during concentration, and the concentration temperature is 50-75° C.; the temperature of the high pressure sterilization is controlled to be 115-135° C.; In step S3, during spray drying, the air inlet temperature is 180-205°C, and the air outlet temperature is 85-105°C.

9. Use of a probiotic Chinese herbal medicine compound composition according to any one of claims 1 to 5 in the preparation of fat-reducing products.

10. The use according to claim 9, characterized in that: The dosage of the probiotic Chinese herbal medicine compound composition is: the dosage of the Chinese herbal medicine compound is 10-24 g / d, and the dosage of the probiotic is 1-6 g / d; and / or, The fat-reducing products include fat-reducing medicines, fat-reducing foods and fat-reducing health products.

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