A liver-benefiting fermented Chinese herbal medicine composition and preparation method thereof

Abstract

The invention discloses a liver-benefiting fermented Chinese herbal medicine composition and a preparation method thereof, which belongs to the technical field of Chinese medicine fermentation, and comprises a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is (1500-2500): (10-30); the Chinese herbal medicine composition is composed of the following components: Artemisia capillaris, Astragalus, green tea extract, Plantago, mint, Coix seeds, safflower, borneol, wolfberry, Lycium barbarum, Lysimachia chinensis, honeysuckle and pumpkin; the probiotic composition is composed of the following components: Bacillus amyloliquefaciens, Bacillus subtilis, Lactobacillus plantarum, Bifidobacterium, pectinase and β-glucanase. The problem of low bioavailability and unstable efficacy of traditional Chinese medicine preparations is solved. It has the advantage of promoting the release and absorption of effective ingredients.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicine fermentation, and in particular to a liver-benefiting fermented Chinese herbal medicine composition and a preparation method thereof. Background Art

[0002] With the changes in modern lifestyles, such as unhealthy eating habits and lack of exercise, the incidence of liver disease continues to rise. Therefore, there is a huge market demand for the development of products with liver protection functions. Chinese herbal medicine has a long history and rich clinical experience. Many Chinese herbal medicines have been proven to have liver protection effects and have also been confirmed to have significant liver protection effects in modern scientific research. These Chinese herbal medicines work through multiple mechanisms, including anti-oxidation, anti-inflammatory, and promoting liver cell repair.

[0003] However, although traditional Chinese herbal medicines have been widely used in clinical practice, there are still some problems in their actual application: (1) Unstable efficacy: Due to the complex composition of Chinese herbal medicines, different extraction processes will cause fluctuations in efficacy. For example, the content and activity of the active ingredients of the same herbal medicine may be different under different extraction conditions, thus affecting the final therapeutic effect. (2) Low bioavailability: Some Chinese herbal medicine ingredients are difficult to be absorbed by the human body, resulting in low bioavailability. For example, some macromolecular compounds are not easy to pass through the intestinal wall into the blood circulation, which limits their efficacy. (3) Potential side effects: Some unprocessed Chinese herbal medicines may have certain side effects. For example, some Chinese herbal medicines contain toxic ingredients that may cause harm to the human body if not properly processed.

[0004] As an emerging technical means, modern fermentation technology can largely solve the above problems. Through the fermentation of microorganisms, the effective ingredients of Chinese herbal medicines can be enhanced, their bioavailability can be improved, and beneficial metabolites such as organic acids and vitamins can be produced, thereby enhancing their health effects. In addition, fermentation technology can also transform complex Chinese herbal medicine ingredients into a form that is more easily absorbed by the human body through enzymatic hydrolysis and other methods, thereby improving the efficacy of the medicine.

[0005] Therefore, there is an urgent need for an efficient liver-benefiting fermented Chinese herbal medicine composition and a preparation method thereof to solve the problems of unstable efficacy and low bioavailability of traditional Chinese herbal medicines, ensure product quality and safety, reduce unnecessary side effects, and provide people with more and better liver health options. Summary of the invention

[0006] To this end, the present invention provides a liver-benefiting fermented Chinese herbal medicine composition and a preparation method thereof to solve the problems of low bioavailability and unstable efficacy of traditional Chinese medicine preparations. In order to achieve the above object, the present invention provides the following technical solutions:

[0007] According to a first aspect of the present invention, there is provided a liver-benefiting fermented Chinese herbal medicine composition, comprising a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is (1500-2500): (10-30);

[0008] The Chinese herbal medicine composition is composed of the following components in parts by weight:

[0009] 20-60 parts of Artemisia capillaris, 50-80 parts of Astragalus, 10-20 parts of green tea extract, 10-30 parts of Plantago, 10-30 parts of Peppermint, 6-20 parts of Coix seeds, 5-18 parts of Carthamus tinctorius, 5-15 parts of Borneolum, 10-20 parts of Lycium barbarum, 10-20 parts of Lysimachia chinensis, 10-20 parts of Honeysuckle and 10-20 parts of Pumpkin;

[0010] The probiotic composition is composed of the following components in parts by weight:

[0011] 1-5 parts of Bacillus amyloliquefaciens, 1-5 parts of Bacillus subtilis, 0.5-3 parts of Lactobacillus plantarum, 1.5-4.5 parts of Bifidobacterium, 0.5-3 parts of pectinase and 0.2-2 parts of β-glucanase;

[0012] The name of the Bacillus amyloliquefaciens is HMJZ-B1003A, the classification name is Bacillus amyloliquefaciens, the preservation number is CCTCC NO: M 20191088, the preservation date is December 23, 2019, the preservation unit is China Center for Type Culture Collection, and the preservation address is Wuhan University, Wuhan, China; the name of the Bacillus subtilis is HMJZ-B1005S, the classification name is Bacillus subtilis, the preservation number is CCTCC NO: M 2020237, the preservation date is June 28, 2020, the preservation unit is China Center for Type Culture Collection, and the preservation address is Wuhan University, Wuhan, China.

[0013] According to a second aspect of the present invention, a method for preparing a liver-benefiting fermented Chinese herbal medicine composition is provided, which is used to prepare the liver-benefiting fermented Chinese herbal medicine composition, comprising the following steps:

[0014] S1. Preparation of primary extract: According to the formula of the herbal composition, weigh each component and prepare a primary extract;

[0015] S2. Preparing a secondary extract: heating the primary extract to enzymatically hydrolyze the primary extract to prepare a secondary extract;

[0016] S3. Preparation of probiotic fermentation broth: Bacillus amyloliquefaciens, Bacillus subtilis, Lactobacillus plantarum and Bifidobacterium were fermented and mixed to obtain a probiotic fermentation broth;

[0017] S4 inoculation and multi-stage fermentation: The secondary extract was inoculated with the probiotic composition fermentation broth obtained in S3 and cultured and fermented to obtain a fermentation broth;

[0018] S5. Inactivation and sealed anaerobic fermentation.

[0019] Furthermore, the S1 comprises the following specific steps:

[0020] The Chinese medicine composition is weighed, mixed and crushed according to the formula, and then 1-2 times the volume of water is added to the Chinese medicine composition for soaking for 6-10 hours. The soaked Chinese medicine composition is extracted through an extraction device to obtain an extract, and the extract is sterilized by ultrafiltration.

[0021] Furthermore, the S2 comprises the following specific steps:

[0022] Heat the mixed medicinal material extract, then add pectinase and heat to 35-60℃ for enzymatic hydrolysis for 15-30min. Use iodine solution for detection. When the medicinal solution does not turn blue when exposed to iodine solution, the enzymatic hydrolysis is completed. Then heat to 100℃ again to inactivate the enzyme for 10-20min. Add impurity remover, stir evenly, let stand for 2h and filter to obtain the secondary extract for use.

[0023] Furthermore, S3 includes the following specific steps:

[0024] S31. Prepare a seed solution of Bacillus amyloliquefaciens, inoculate the seed solution of Bacillus amyloliquefaciens into a sterilized medium A for fermentation, the culture temperature is 35-38 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200-230rpm, the fermentation culture is 72-96h, and the spore formation rate is cultured to more than 90%;

[0025] The culture medium A comprises the following components in the following concentrations: 3-8 g / L glucose, 25-45 g / L corn starch, 30-50 g / L soybean meal, 6-12 g / L yeast powder, 5-10 g / L peptone, 1-5 g / L calcium carbonate, 1-3 g / L sodium chloride, 0.8-8 g / L magnesium sulfate, 1-4 g / L potassium hydrogen phosphate and 0.2-0.5 g / L manganese sulfate, and the pH is adjusted to 7-7.2;

[0026] The seed liquid of Bacillus amyloliquefaciens comprises the following preparation steps: picking a single colony from the preserved strains, inoculating it into a sterilized seed culture medium for cultivation, the cultivation temperature is 35-40° C., the dissolved oxygen is not less than 20%, and the spore formation rate is above 90%;

[0027] The seed culture medium comprises the following components in the following concentrations: 5-15 g / L yeast extract, 10-20 g / L peptone and 1-5 g / L sodium chloride;

[0028] S32. Prepare a seed solution of Bacillus subtilis, inoculate the seed solution of Bacillus subtilis into a sterilized medium B for fermentation, the culture temperature is 35-38 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 150-200 rpm, the fermentation culture is 8-16h, and the spore formation rate is cultured to more than 90%;

[0029] The culture medium B comprises the following components in the following concentrations: glucose 2-7 g / L, corn starch 20-40 g / L, soybean meal 30-50 g / L, yeast powder 3-8 g / L, peptone 3-8 g / L, calcium carbonate 1-5 g / L, sodium chloride 1-5 g / L, urea 1-4 g / L, magnesium sulfate 0.5-1 g / L, dipotassium hydrogen phosphate 2-8 g / L, manganese sulfate 0.1-0.3 g / L, folate 0.1-0.3 g / L, the balance is water, and the pH is adjusted to 6-7;

[0030] The seed liquid of Bacillus subtilis comprises the following steps: picking a single colony from the preserved strain, streaking and inoculating it in a sterilized seed culture medium for culturing, the culture temperature is 35-38°C, the dissolved oxygen is not less than 20%, static culture is carried out for 5-7 days, and culturing is carried out until the spore formation rate is above 90%;

[0031] The seed culture medium comprises the following components in the following concentrations: 2-5 g / L beef extract, 3-8 g / L sodium chloride, 8-12 g / L peptone, 15-20 g / L agar, the remainder being water, and the pH is adjusted to 7-7.5;

[0032] S33 Lactobacillus plantarum anaerobic fermentation: After autoclaving, the culture medium C was heated to 30-40 ℃, pH was adjusted to 5-5.5, Lactobacillus plantarum powder was added, and the mixture was stirred and then cultured in an anaerobic closed environment for 8-10h to obtain Lactobacillus plantarum culture after expansion and proliferation;

[0033] The culture solution C includes the following components in the following concentrations: peptone 10-20 g / L, beef extract 5-10 g / L, yeast extract 5-10 g / L, glucose 20-25 g / L, Tween-80 0.5-1.5 g / L, dipotassium hydrogen phosphate 1-3 g / L, sodium acetate 4-6 g / L, magnesium sulfate heptahydrate 0.2-0.6 g / L and manganese sulfate 0.1-0.3 g / L;

[0034] S34 Bifidobacterium anaerobic fermentation: After autoclaving, the culture medium D was inoculated into the fermentation medium and cultured anaerobically at 35-39 ° C for 36-60h. After the fermentation, the solid impurities were removed by a disc centrifuge, and then the soluble impurities were removed by microfiltration to obtain the fermentation supernatant;

[0035] The culture solution D includes the following components in the following concentrations: glucose 1-12 g / L, tryptone 1-14.5 g / L, inorganic salts 2-16.5 g / L, Tween-80 0.2-2.5 g / L, and ascorbic acid 0.1-1.5 g / L;

[0036] S35. Mix the expanded and multiplied bacterial liquids obtained in S31, S32, S33 and S34 to obtain a probiotic composition fermented bacterial liquid.

[0037] Furthermore, the S4 comprises the following specific steps:

[0038] When the temperature of the secondary extract drops to 25-35°C, the probiotic composition fermentation liquid is inoculated and cultured until the pH value drops to 5.5-6.5, glucose is added and sterilized at high temperature, and when the temperature drops to 35-40°C, the fermentation is continued for 20-24 hours. When the pH value drops to 4.1-4.5, the fermentation temperature is lowered to 15-25°C, and the fermentation is continued until the pH value drops to 3.5-3.8, and the fermentation is terminated.

[0039] Furthermore, S5 comprises the following specific steps: adding β-glucanase to the fermentation broth and mixing well, maintaining a constant temperature of 30-60° C. for 24-48 hours, and performing sealed anaerobic fermentation for 36-72 hours to obtain the final fermented Chinese herbal medicine composition.

[0040] Furthermore, the impurity remover in S2 is activated carbon, the addition amount is 0.5 g / L, and the addition amount of pectinase is 3 wt % of the medicinal material extract.

[0041] Furthermore, the amount of β-glucanase added in S5 is 5wt% of the fermentation broth.

[0042] According to a third aspect of the present invention, there is provided a use of the liver-benefiting fermented Chinese herbal medicine composition in the preparation of a liver-benefiting medicine.

[0043] The present invention has the following advantages:

[0044] 1. The various Chinese herbal medicine ingredients such as Artemisia capillaris, Astragalus membranaceus, and green tea extract in the formula of this application work together and play a role together, especially the addition of borneol and moneywort. The former has the effects of awakening the mind, clearing away heat and relieving pain, and can effectively neutralize the heat toxins of the liver and inhibit inflammatory reactions; the latter has the effects of clearing the liver and gallbladder, detoxifying and reducing swelling. These ingredients can enhance the comprehensive efficacy of the drug and improve liver function. The addition of probiotics such as Bacillus amyloliquefaciens and Bacillus subtilis can not only help digestion and absorption, but also promote intestinal health, enhance the body's absorption and utilization of Chinese herbal medicine ingredients, and further enhance the comprehensive efficacy of the drug. Bacillus amyloliquefaciens can produce a variety of antimicrobial peptides and enzymes, which help to improve the balance of intestinal microecology; Bacillus subtilis can enhance immunomodulatory effects and improve the body's resistance to disease.

[0045] 2. This application uses a multi-stage fermentation technology to first activate probiotics using the nutrients in the Chinese herbal extract, and then through temperature control and pH adjustment at different stages to ensure that the probiotics grow and reproduce under optimal conditions, and finally form a fermentation product with a high spore rate. This multi-stage fermentation process not only improves the quality of the fermentation product, but also increases the bioavailability of the drug ingredients. The use of pectinase and β-glucanase effectively decomposes the macromolecular substances in the Chinese herbal extract and improves the availability of nutrients. In particular, the later addition of β-glucanase further improves the purity and stability of the fermentation product, ensuring the quality and safety of the final product.

[0046] 3. The technical solution of this application comprehensively considers the effective ingredients of Chinese herbal medicines and the metabolites of probiotics. This technical solution can improve the bioavailability of drugs and make the drug ingredients more easily absorbed and utilized by the human body, thereby achieving better therapeutic effects. The metabolites produced by probiotics during the fermentation process help improve the intestinal microecological environment and promote the absorption of nutrients, thereby further improving the comprehensive efficacy of drugs; the synergistic effect of Bacillus amyloliquefaciens and Bacillus subtilis in promoting intestinal health gives this technical solution a unique advantage in the preparation of liver-benefiting drugs. DETAILED DESCRIPTION

[0047] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are 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 ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0048] The present invention provides a liver-benefiting fermented Chinese herbal medicine composition, comprising a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is (1500-2500): (10-30);

[0049] The Chinese herbal medicine composition is composed of the following components in parts by weight:

[0050] 20-60 parts of Artemisia capillaris, 50-80 parts of Astragalus, 10-20 parts of green tea extract, 10-30 parts of Plantago, 10-30 parts of Peppermint, 6-20 parts of Coix seeds, 5-18 parts of Carthamus tinctorius, 5-15 parts of Borneolum, 10-20 parts of Lycium barbarum, 10-20 parts of Lysimachia chinensis, 10-20 parts of Honeysuckle and 10-20 parts of Pumpkin;

[0051] The probiotic composition is composed of the following components in parts by weight:

[0052] 1-5 parts of Bacillus amyloliquefaciens, 1-5 parts of Bacillus subtilis, 0.5-3 parts of Lactobacillus plantarum, 1.5-4.5 parts of Bifidobacterium, 0.5-3 parts of pectinase and 0.2-2 parts of β-glucanase;

[0053] The name of Bacillus amyloliquefaciens is HMJZ-B1003A, the classification name is Bacillusamyloliquefaciens, the deposit number is CCTCC NO: M 20191088, the deposit date is December 23, 2019, the deposit unit is China Center for Type Culture Collection, and the deposit address is Wuhan University, Wuhan, China; the name of Bacillus subtilis is HMJZ-B1005S, the classification name is Bacillus subtilis, the deposit number is CCTCC NO: M 2020237, the deposit date is June 28, 2020, the deposit unit is China Center for Type Culture Collection, and the deposit address is Wuhan University, Wuhan, China. The above two strains have been disclosed in the applicant's previous patent "A microbial fermented herbal atomized ecological liquid and its preparation method and application" (patent application number: CN202410581637.8).

[0054] The present invention provides a method for preparing a liver-benefiting fermented Chinese herbal medicine composition, which is used to prepare the liver-benefiting fermented Chinese herbal medicine composition, comprising the following steps:

[0055] S1. Preparation of primary extract: According to the formula of the herbal composition, weigh each component and prepare a primary extract;

[0056] S2. Preparing a secondary extract: heating the primary extract to enzymatically hydrolyze the primary extract to prepare a secondary extract;

[0057] S3. Preparation of probiotic composition fermentation broth: Bacillus amyloliquefaciens, Bacillus subtilis, Lactobacillus plantarum, Bifidobacterium were fermented and mixed to obtain a probiotic composition fermentation broth;

[0058] S4 inoculation and multi-stage fermentation: The secondary extract was inoculated with the probiotic composition fermentation broth obtained in S3 and cultured and fermented to obtain a fermentation broth;

[0059] S5. Inactivation and sealed anaerobic fermentation.

[0060] Among them, S1 includes the following specific steps:

[0061] The Chinese medicine composition is weighed, mixed and crushed according to the formula, and then 1-2 times the volume of water is added to the Chinese medicine composition for soaking for 6-10 hours. The soaked Chinese medicine composition is extracted through an extraction device to obtain an extract, and the extract is sterilized by ultrafiltration.

[0062] Among them, S2 includes the following specific steps:

[0063] Heat the mixed medicinal material extract, then add pectinase and heat to 35-60℃ for enzymatic hydrolysis for 15-30min. Use iodine solution for detection. When the medicinal solution does not turn blue when exposed to iodine solution, the enzymatic hydrolysis is completed. Then heat to 100℃ again to inactivate the enzyme for 10-20min. Add impurity remover, stir evenly, let stand for 2h and filter to obtain the secondary extract for use.

[0064] Among them, S3 includes the following specific steps:

[0065] S31. Prepare a seed solution of Bacillus amyloliquefaciens, inoculate the seed solution of Bacillus amyloliquefaciens into a sterilized medium A for fermentation, the culture temperature is 35-38 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200-230 rpm, the fermentation culture is 72-96h, and the spore formation rate is cultured to more than 90%;

[0066] Medium A includes the following components in the following concentrations: glucose 3-8 g / L, corn starch 25-45 g / L, soybean meal 30-50 g / L, yeast powder 6-12 g / L, peptone 5-10 g / L, calcium carbonate 1-5 g / L, sodium chloride 1-3 g / L, magnesium sulfate 0.8-8 g / L, potassium dihydrogen phosphate 1-4 g / L and manganese sulfate 0.2-0.5 g / L, and the pH is adjusted to 7-7.2;

[0067] The seed solution of Bacillus amyloliquefaciens comprises the following steps: picking a single colony from the preserved strain, inoculating it into a sterilized seed culture medium for cultivation, the cultivation temperature is 35-40° C., the dissolved oxygen is not less than 20%, and the spore formation rate is above 90%;

[0068] The seed culture medium includes the following components at the following concentrations: yeast extract 5-15 g / L, peptone 10-20 g / L and sodium chloride 1-5 g / L;

[0069] S32. Preparation of Bacillus subtilis seed solution, the seed solution of Bacillus subtilis was inoculated into sterilized medium B for fermentation, the culture temperature was 35-38 ° C, the dissolved oxygen was not less than 20%, the stirring speed was 150-200 rpm, the fermentation culture was 8-16h, and the spore formation rate was cultured to more than 90%;

[0070] Medium B includes the following components in the following concentrations: glucose 2-7 g / L, corn starch 20-40 g / L, soybean meal 30-50 g / L, yeast powder 3-8 g / L, peptone 3-8 g / L, calcium carbonate 1-5 g / L, sodium chloride 1-5 g / L, urea 1-4 g / L, magnesium sulfate 0.5-1 g / L, potassium dihydrogen phosphate 2-8 g / L, manganese sulfate 0.1-0.3 g / L, folate 0.1- 0.3 g / L, the balance is water, and the pH is adjusted to 6-7;

[0071] The seed liquid of Bacillus subtilis comprises the following steps: picking a single colony from the preserved strain, streaking and inoculating it in a sterilized seed culture medium for culturing, the culture temperature is 35-38°C, the dissolved oxygen is not less than 20%, static culture is carried out for 5-7 days, and culturing is carried out until the spore formation rate is above 90%;

[0072] The seed culture medium includes the following components in the following concentrations: beef extract 2-5 g / L, sodium chloride 3-8 g / L, peptone 8-12 g / L, agar 15-20 g / L, the balance is water, and the pH is adjusted to 7-7.5;

[0073] S33 Lactobacillus plantarum anaerobic fermentation: After autoclaving, the culture medium C was heated to 30-40 ℃, pH was adjusted to 5-5.5, Lactobacillus plantarum powder was added, and the mixture was stirred and then cultured in an anaerobic closed environment for 8-10h to obtain Lactobacillus plantarum culture after expansion and proliferation;

[0074] The culture medium C includes the following components in the following concentrations: peptone 10-20 g / L, beef extract 5-10 g / L, yeast extract 5-10 g / L, glucose 20-25 g / L, Tween-80 0.5-1.5 g / L, dipotassium hydrogen phosphate 1-3 g / L, sodium acetate 4-6 g / L, magnesium sulfate heptahydrate 0.2-0.6 g / L and manganese sulfate 0.1-0.3 g / L;

[0075] S34 Bifidobacterium anaerobic fermentation: After autoclaving, the culture medium D was inoculated into the fermentation medium and cultured anaerobically at 35-39 ° C for 36-60h. After the fermentation, the solid impurities were removed by a disc centrifuge, and then the soluble impurities were removed by microfiltration to obtain the fermentation supernatant;

[0076] The culture medium D includes the following components at the following concentrations: glucose 1-12 g / L, tryptone 1-14.5 g / L, inorganic salts 2-16.5 g / L, Tween-80 0.2-2.5 g / L, and ascorbic acid 0.1-1.5 g / L;

[0077] S35. Mix the expanded and multiplied bacterial liquids obtained in S31, S32, S33 and S34 to obtain a probiotic composition fermented bacterial liquid.

[0078] Among them, S4 includes the following specific steps:

[0079] When the temperature of the secondary extract drops to 25-35°C, the probiotic composition fermentation liquid is inoculated and cultured until the pH value drops to 5.5-6.5, glucose is added and sterilized at high temperature, and when the temperature drops to 35-40°C, the fermentation is continued for 20-24 hours. When the pH value drops to 4.1-4.5, the fermentation temperature is lowered to 15-25°C, and the fermentation is continued until the pH value drops to 3.5-3.8, and the fermentation is terminated.

[0080] Among them, S5 includes the following specific steps: adding β-glucanase to the fermentation broth and mixing it evenly, keeping it at a constant temperature of 30-60°C for 24-48 hours, and then sealing and anaerobic fermenting it for 36-72 hours to obtain the final fermented Chinese herbal medicine composition.

[0081] Specifically, the impurity remover in S2 is activated carbon, the addition amount is 0.5 g / L, and the addition amount of pectinase is 3 wt % of the medicinal material extract.

[0082] Specifically, the amount of β-glucanase added in S5 is 5 wt % of the fermentation broth.

[0083] The present invention provides an application of the liver-benefiting fermented Chinese herbal medicine composition in the preparation of a liver-benefiting medicine. Example

[0084] This embodiment provides a liver-benefiting fermented Chinese herbal medicine composition, comprising a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is 1500:10;

[0085] The Chinese herbal medicine composition is composed of the following components in parts by weight:

[0086] Artemisia capillaris 40g, astragalus 65g, green tea extract 15g, plantain 20g, peppermint 20g, coix seed 13g, safflower 12g, borneol 12g, wolfberry 15g, desmodium 15g, honeysuckle 15g and pumpkin 15g;

[0087] The probiotic composition is composed of the following components in parts by weight:

[0088] Bacillus amyloliquefaciens 5g, Bacillus subtilis 5g, Lactobacillus plantarum 3g, Bifidobacterium 3g, pectinase 2g and β-glucanase 1.5g.

[0089] This embodiment provides a method for preparing a liver-benefiting fermented Chinese herbal medicine composition, comprising the following steps:

[0090] S1. Weigh, mix and crush the Chinese medicine composition according to the formula, then add 1.5 times the volume of water to the Chinese medicine composition and soak it for 10 hours, extract the soaked Chinese medicine composition through an extraction device to obtain an extract, and ultrafilter the extract to sterilize it.

[0091] S2. Heat the mixed medicinal material extract, then add 3wt% pectinase to the medicinal material extract, heat to 45℃ for enzymolysis for 20min, use iodine solution for detection, and the enzymolysis is completed when the medicinal solution does not turn blue when exposed to iodine solution. Then heat to 100℃ again to inactivate the enzyme for 15min, add 0.5g / L activated carbon, stir evenly, let stand for 2h, and then filter to obtain the secondary extract for use.

[0092] S3 includes the following specific steps:

[0093] S31. Prepare a seed solution of Bacillus amyloliquefaciens, inoculate the seed solution of Bacillus amyloliquefaciens into a sterilized medium A for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 72h, and the spore formation rate is cultured to more than 90%;

[0094] Medium A includes the following components at the following concentrations: glucose 6 g / L, corn starch 40 g / L, soybean meal 35 g / L, yeast powder 9 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 2 g / L, magnesium sulfate 0.8 g / L, potassium hydrogen phosphate 2 g / L, and manganese sulfate 0.3 g / L, and the pH is adjusted to 7-7.2;

[0095] The seed solution of Bacillus amyloliquefaciens comprises the following steps: picking a single colony from the preserved strain, inoculating it into a sterilized seed culture medium for cultivation, the cultivation temperature is 35-40° C., the dissolved oxygen is not less than 20%, and the spore formation rate is above 90%;

[0096] The seed culture medium includes the following components at the following concentrations: yeast extract 10 g / L, peptone 20 g / L, and sodium chloride 5 g / L;

[0097] S32. Prepare a seed solution of Bacillus subtilis, inoculate the seed solution of Bacillus subtilis into a sterilized medium B for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 12h, and the spore formation rate is cultured to more than 90%;

[0098] Medium B includes the following components in the following concentrations: glucose 5 g / L, corn starch 30 g / L, soybean meal 40 g / L, yeast powder 8 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 3 g / L, urea 2 g / L, magnesium sulfate 0.8 g / L, potassium dihydrogen phosphate 3 g / L, manganese sulfate 0.3 g / L, folate 0.2 g / L, the balance is water, and the pH is adjusted to 6-7;

[0099] The seed liquid of Bacillus subtilis comprises the following steps: picking a single colony from the preserved strain, streaking and inoculating it in a sterilized seed culture medium for culturing, the culture temperature is 37° C., the dissolved oxygen is not less than 20%, static culture is carried out for 7 days, and spore formation rate is above 90%;

[0100] The seed culture medium includes the following components in the following concentrations: 3 g / L beef extract, 5 g / L sodium chloride, 10 g / L peptone, 18 g / L agar, the balance being water, and the pH is adjusted to 7-7.5;

[0101] S33. Anaerobic fermentation of Lactobacillus plantarum: After autoclaving, the culture medium C was heated to 30 ° C, the pH was adjusted to 5.5, Lactobacillus plantarum powder was added, and the mixture was stirred and then cultured in an anaerobic closed environment for 9.6 h to obtain a Lactobacillus plantarum culture solution after multiplication and proliferation;

[0102] Culture medium C includes the following components in the following concentrations: peptone 15 g / L, beef extract 6 g / L, yeast extract 6 g / L, glucose 22 g / L, Tween-80 1 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 4.5 g / L, magnesium sulfate heptahydrate 0.4 g / L, and manganese sulfate 0.2 g / L.

[0103] S34 Bifidobacterium anaerobic fermentation: After autoclaving, the culture medium D was inoculated into the fermentation medium and cultured anaerobically at 37 ° C for 48 h. After the fermentation, the solid impurities were removed by a disc centrifuge, and then the soluble impurities were removed by microfiltration to obtain the fermentation supernatant;

[0104] The culture medium D includes the following components at the following concentrations: glucose 6 g / L, tryptone 7.75 g / L, inorganic salt 9 g / L, Tween-80 1.25 ml / L, and ascorbic acid 0.8 g / L.

[0105] S35. Mix the expanded and multiplied bacterial liquids obtained in S31, S32, S33 and S34 to obtain a probiotic composition fermented bacterial liquid.

[0106] S4. When the temperature of the secondary extract drops to 35°C, inoculate the fermentation broth of the probiotic composition and culture it until the pH value drops to 5.5-6.5, add glucose and sterilize it at high temperature, and when the temperature drops to 30°C, continue fermenting for 20 hours. When the pH value drops to 4.1-4.5, lower the fermentation temperature to 20°C, continue fermenting until the pH value drops to 3.5-3.8, and end the fermentation.

[0107] S5: adding 5 wt% of β-glucanase to the fermentation broth, mixing well, keeping a constant temperature of 60° C. for 48 hours, and then sealing and performing anaerobic fermentation for 72 hours to obtain the final fermented Chinese herbal medicine composition. Example

[0108] This embodiment provides a liver-benefiting fermented Chinese herbal medicine composition, comprising a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is 2000:20;

[0109] The Chinese herbal medicine composition is composed of the following components in parts by weight:

[0110] Artemisia capillaris 40g, astragalus 65g, green tea extract 15g, plantain 20g, peppermint 20g, coix seed 13g, safflower 12g, borneol 12g, wolfberry 15g, desmodium 15g, honeysuckle 15g and pumpkin 15g;

[0111] The probiotic composition is composed of the following components in parts by weight:

[0112] Bacillus amyloliquefaciens 5g, Bacillus subtilis 5g, Lactobacillus plantarum 3g, Bifidobacterium 3g, pectinase 2g and β-glucanase 1.5g.

[0113] This embodiment provides a method for preparing a liver-benefiting fermented Chinese herbal medicine composition, comprising the following steps:

[0114] S1. Weigh, mix and crush the Chinese medicine composition according to the formula, then add 1.5 times the volume of water to the Chinese medicine composition and soak it for 10 hours, extract the soaked Chinese medicine composition through an extraction device to obtain an extract, and ultrafilter the extract to sterilize it.

[0115] S2. Heat the mixed medicinal material extract, then add 3wt% pectinase to the medicinal material extract, heat to 45℃ for enzymolysis for 20min, use iodine solution for detection, and the enzymolysis is completed when the medicinal solution does not turn blue when exposed to iodine solution. Then heat to 100℃ again to inactivate the enzyme for 15min, add 0.5g / L activated carbon, stir evenly, let stand for 2h, and then filter to obtain the secondary extract for use.

[0116] S3 includes the following specific steps:

[0117] S31. Prepare a seed solution of Bacillus amyloliquefaciens, inoculate the seed solution of Bacillus amyloliquefaciens into a sterilized medium A for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 72h, and the spore formation rate is cultured to more than 90%;

[0118] Medium A includes the following components at the following concentrations: glucose 6 g / L, corn starch 40 g / L, soybean meal 35 g / L, yeast powder 9 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 2 g / L, magnesium sulfate 0.8 g / L, potassium hydrogen phosphate 2 g / L, and manganese sulfate 0.3 g / L, and the pH is adjusted to 7-7.2;

[0119] The seed solution of Bacillus amyloliquefaciens comprises the following steps: picking a single colony from the preserved strain, inoculating it into a sterilized seed culture medium for cultivation, the cultivation temperature is 35-40° C., the dissolved oxygen is not less than 20%, and the spore formation rate is above 90%;

[0120] The seed culture medium includes the following components at the following concentrations: yeast extract 10 g / L, peptone 20 g / L, and sodium chloride 5 g / L;

[0121] S32. Prepare a seed solution of Bacillus subtilis, inoculate the seed solution of Bacillus subtilis into a sterilized medium B for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 12h, and the spore formation rate is cultured to more than 90%;

[0122] Medium B includes the following components in the following concentrations: glucose 5 g / L, corn starch 30 g / L, soybean meal 40 g / L, yeast powder 8 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 3 g / L, urea 2 g / L, magnesium sulfate 0.8 g / L, potassium dihydrogen phosphate 3 g / L, manganese sulfate 0.3 g / L, folate 0.2 g / L, the balance is water, and the pH is adjusted to 6-7;

[0123] The seed liquid of Bacillus subtilis comprises the following steps: picking a single colony from the preserved strain, streaking and inoculating it in a sterilized seed culture medium for culturing, the culture temperature is 37° C., the dissolved oxygen is not less than 20%, static culture is carried out for 7 days, and spore formation rate is above 90%;

[0124] The seed culture medium includes the following components in the following concentrations: 3 g / L beef extract, 5 g / L sodium chloride, 10 g / L peptone, 18 g / L agar, the balance being water, and the pH is adjusted to 7-7.5;

[0125] S33. Anaerobic fermentation of Lactobacillus plantarum: After autoclaving, the culture medium C was heated to 30 ° C, the pH was adjusted to 5.5, Lactobacillus plantarum powder was added, and the mixture was stirred and then cultured in an anaerobic closed environment for 9.6 h to obtain a Lactobacillus plantarum culture solution after multiplication and proliferation;

[0126] Culture medium C includes the following components in the following concentrations: peptone 15 g / L, beef extract 6 g / L, yeast extract 6 g / L, glucose 22 g / L, Tween-80 1 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 4.5 g / L, magnesium sulfate heptahydrate 0.4 g / L, and manganese sulfate 0.2 g / L.

[0127] S34 Bifidobacterium anaerobic fermentation: After autoclaving, the culture medium D was inoculated into the fermentation medium and cultured anaerobically at 37 ° C for 48 h. After the fermentation, the solid impurities were removed by a disc centrifuge, and then the soluble impurities were removed by microfiltration to obtain the fermentation supernatant;

[0128] The culture medium D includes the following components at the following concentrations: glucose 6 g / L, tryptone 7.75 g / L, inorganic salt 9 g / L, Tween-80 1.25 ml / L, and ascorbic acid 0.8 g / L.

[0129] S35. Mix the expanded and multiplied bacterial liquids obtained in S31, S32, S33 and S34 to obtain a probiotic composition fermented bacterial liquid.

[0130] S4. When the temperature of the secondary extract drops to 35°C, inoculate the fermentation broth of the probiotic composition and culture it until the pH value drops to 5.5-6.5, add glucose and sterilize it at high temperature, and when the temperature drops to 30°C, continue fermenting for 20 hours. When the pH value drops to 4.1-4.5, lower the fermentation temperature to 20°C, continue fermenting until the pH value drops to 3.5-3.8, and end the fermentation.

[0131] S5. Add 5 wt% of β-glucanase to the fermentation broth, mix well, keep constant temperature at 60°C for 48 hours, and then seal and perform anaerobic fermentation for 72 hours to obtain the final fermented Chinese herbal medicine composition. Example

[0132] This embodiment provides a liver-benefiting fermented Chinese herbal medicine composition, comprising a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is 2500:30;

[0133] The Chinese herbal medicine composition is composed of the following components in parts by weight:

[0134] Artemisia capillaris 40g, astragalus 65g, green tea extract 15g, plantain 20g, peppermint 20g, coix seed 13g, safflower 12g, borneol 12g, wolfberry 15g, desmodium 15g, honeysuckle 15g and pumpkin 15g;

[0135] The probiotic composition is composed of the following components in parts by weight:

[0136] Bacillus amyloliquefaciens 5g, Bacillus subtilis 5g, Lactobacillus plantarum 3g, Bifidobacterium 3g, pectinase 2g and β-glucanase 1.5g.

[0137] This embodiment provides a method for preparing a liver-benefiting fermented Chinese herbal medicine composition, comprising the following steps:

[0138] S1. Weigh, mix and crush the Chinese medicine composition according to the formula, then add 1.5 times the volume of water to the Chinese medicine composition and soak it for 10 hours, extract the soaked Chinese medicine composition through an extraction device to obtain an extract, and ultrafilter the extract to sterilize it.

[0139] S2. Heat the mixed medicinal material extract, then add 3wt% pectinase to the medicinal material extract, heat to 45℃ for enzymolysis for 20min, use iodine solution for detection, and the enzymolysis is completed when the medicinal solution does not turn blue when exposed to iodine solution. Then heat to 100℃ again to inactivate the enzyme for 15min, add 0.5g / L activated carbon, stir evenly, let stand for 2h, and then filter to obtain the secondary extract for use.

[0140] S3 includes the following specific steps:

[0141] S31. Prepare a seed solution of Bacillus amyloliquefaciens, inoculate the seed solution of Bacillus amyloliquefaciens into a sterilized medium A for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 72h, and the spore formation rate is cultured to more than 90%;

[0142] Medium A includes the following components at the following concentrations: glucose 6 g / L, corn starch 40 g / L, soybean meal 35 g / L, yeast powder 9 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 2 g / L, magnesium sulfate 0.8 g / L, potassium hydrogen phosphate 2 g / L, and manganese sulfate 0.3 g / L, and the pH is adjusted to 7-7.2;

[0143] The seed solution of Bacillus amyloliquefaciens comprises the following steps: picking a single colony from the preserved strain, inoculating it into a sterilized seed culture medium for culturing at a temperature of 37° C. and a dissolved oxygen content of not less than 20%, and culturing until the spore formation rate is above 90%;

[0144] The seed culture medium includes the following components at the following concentrations: yeast extract 10 g / L, peptone 20 g / L, and sodium chloride 5 g / L;

[0145] S32. Prepare a seed solution of Bacillus subtilis, inoculate the seed solution of Bacillus subtilis into a sterilized medium B for fermentation, the culture temperature is 37 ° C, the dissolved oxygen is not less than 20%, the stirring speed is 200 rpm, the fermentation culture is 12h, and the spore formation rate is cultured to more than 90%;

[0146] Medium B includes the following components in the following concentrations: glucose 5 g / L, corn starch 30 g / L, soybean meal 40 g / L, yeast powder 8 g / L, peptone 8 g / L, calcium carbonate 3 g / L, sodium chloride 3 g / L, urea 2 g / L, magnesium sulfate 0.8 g / L, potassium dihydrogen phosphate 3 g / L, manganese sulfate 0.3 g / L, folate 0.2 g / L, the balance is water, and the pH is adjusted to 6-7;

[0147] The seed liquid of Bacillus subtilis comprises the following steps: picking a single colony from the preserved strain, streaking and inoculating it in a sterilized seed culture medium for culturing, the culture temperature is 37° C., the dissolved oxygen is not less than 20%, static culture is carried out for 7 days, and spore formation rate is above 90%;

[0148] The seed culture medium includes the following components in the following concentrations: 3 g / L beef extract, 5 g / L sodium chloride, 10 g / L peptone, 18 g / L agar, the balance being water, and the pH is adjusted to 7-7.5;

[0149] S33. Anaerobic fermentation of Lactobacillus plantarum: After autoclaving, the culture medium C was heated to 30 ° C, the pH was adjusted to 5.5, Lactobacillus plantarum powder was added, and the mixture was stirred and then cultured in an anaerobic closed environment for 9.6 h to obtain a Lactobacillus plantarum culture solution after multiplication and proliferation;

[0150] Culture medium C includes the following components in the following concentrations: peptone 15 g / L, beef extract 6 g / L, yeast extract 6 g / L, glucose 22 g / L, Tween-80 1 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 4.5 g / L, magnesium sulfate heptahydrate 0.4 g / L, and manganese sulfate 0.2 g / L.

[0151] S34 Bifidobacterium anaerobic fermentation: After autoclaving, the culture medium D was inoculated into the fermentation medium and cultured anaerobically at 37 ° C for 48 h. After the fermentation, the solid impurities were removed by a disc centrifuge, and then the soluble impurities were removed by microfiltration to obtain the fermentation supernatant;

[0152] The culture medium D includes the following components at the following concentrations: glucose 6 g / L, tryptone 7.75 g / L, inorganic salt 9 g / L, Tween-80 1.25 ml / L, and ascorbic acid 0.8 g / L.

[0153] S35. Mix the expanded and multiplied bacterial liquids obtained in S31, S32, S33 and S34 to obtain a probiotic composition fermented bacterial liquid.

[0154] S4. When the temperature of the secondary extract drops to 35°C, inoculate the fermentation broth of the probiotic composition and culture it until the pH value drops to 5.5-6.5, add glucose and sterilize it at high temperature, and when the temperature drops to 30°C, continue fermenting for 20 hours. When the pH value drops to 4.1-4.5, lower the fermentation temperature to 20°C, continue fermenting until the pH value drops to 3.5-3.8, and end the fermentation.

[0155] S5. Add 5 wt% of β-glucanase to the fermentation broth, mix well, keep constant temperature at 60°C for 48 hours, and then seal and perform anaerobic fermentation for 72 hours to obtain the final fermented Chinese herbal medicine composition. Example

[0156] The fermented Chinese herbal medicine composition prepared as above is freeze-dried or spray-dried to obtain a solid powder; or appropriate adhesives, wetting agents, other auxiliary materials, etc. are added to make a soft material, which is sieved to form wet granules. The wet granules are dried, sized, compressed, and filled into capsules to form solid dosage forms such as granules, tablets, and capsules; each dose in the solid preparation is 5g.

[0157] Example 5: Drug efficacy evaluation and liver protection effect verification test

[0158] The properties of the granules of the fermented Chinese herbal medicine composition obtained in Examples 1-3 were measured.

[0159] (1) Determination of properties of CCl4 liver injury model

[0160] Experimental animals: 120 healthy Kunming mice, 60 male and 60 female mice, weighing 15-20g respectively, were randomly divided into 5 groups, 3 treatment groups, 1 control group, and 1 blank group, with 24 mice in each group.

[0161] Experimental method: The mice in the three treatment groups were fed with 0.5 g / kg of the granules of the fermented Chinese herbal composition obtained in Examples 1-3 in the morning, noon and evening, respectively, by gavage, and the control group was gavaged with the same amount of physiological saline. The blank group was not given any medicine and no modeling was done. The mice were gavaged three times a day, in the morning, noon and evening, for 7 consecutive days (1 course of treatment). One hour after the last administration, each group was given a 0.1% CCl4 soybean oil complex solution. Blood was collected after fasting for 24 hours, and the serum was separated for testing to measure the serum ALT and AST values, and liver tissue was taken and processed to measure the SOD activity and MDA content.

[0162] (2) Pharmacological experiments on TAA liver injury model

[0163] Experimental animals: 80 healthy Kunming mice, half male and half female, weighing 15-20 g, were randomly divided into 4 groups, including a control group of 20 mice, a treatment group 1 that took the fermented Chinese herbal composition granules prepared in Example 1, a treatment group 2 that took the fermented Chinese herbal composition granules prepared in Example 2, and a treatment group 3 that took the fermented Chinese herbal composition granules prepared in Example 3, each with 20 mice.

[0164] Experimental method: The control group was gavaged with an equal amount of injection water every day, and the three treatment groups were given 0.5 / kg of fermented Chinese herbal medicine composition granules. After 14 days (2 courses of treatment), the three treatment groups and the control group were intraperitoneally injected with thioacetamide (TAA) and fasted but not water. Blood was collected from the eyeballs for 24 hours, and the serum was separated for testing to determine the ALT and AST values.

[0165] Results Analysis

[0166] (1) Analysis of the results of CCl4 liver injury model

[0167] According to the experimental operation of property determination (1), serum ALT and AST values ​​were measured, and liver tissue was taken and processed to determine SOD activity and MDA content. The results are shown in Table 1 below:

[0168] Table 1 Result analysis data of CCl4 liver injury model

[0169]

[0170] According to the test results in Table 1, (1) the ALT level in treatment group 2 was the lowest, followed by treatment group 1 and treatment group 3, which were significantly lower than the control group; this indicates that treatment group 2 had the best effect in reducing ALT levels, indicating that the degree of liver cell damage was minimal; (2) the AST level in treatment group 2 was the lowest, followed by treatment group 1 and treatment group 3, which were significantly lower than the control group. This indicates that treatment group 2 had the best effect in reducing AST levels, indicating that the degree of liver cell damage was minimal. (3) the MDA level in treatment group 2 was the lowest, followed by treatment group 1 and treatment group 3, which were significantly lower than the control group; this indicates that treatment group 2 had the best effect in reducing lipid peroxidation and had a strong antioxidant capacity. (4) the SOD level in treatment group 2 was the highest, followed by treatment group 1 and treatment group 3, which were significantly higher than the control group; this indicates that treatment group 2 had the best effect in enhancing antioxidant capacity.

[0171] In summary, the fermented Chinese herbal medicine composition provided by the present application has a significant inhibitory effect on the increase of ALT, AST and MDA in mice caused by CCl4, and has a significant effect on causing a decrease in SOD activity, indicating that the fermented Chinese herbal medicine composition provided by the present application has a significant improvement effect on liver damage caused by CCl4. In particular, the overall effect of treatment group 2 is the best, with significant effects in reducing liver damage, reducing lipid peroxidation and enhancing antioxidant capacity.

[0172] (2) Analysis of results of TAA liver injury model

[0173] According to the experimental operation of property determination (2), the ALT value and AST value were determined. The results are shown in Table 2 below:

[0174] Table 2 Result analysis data of TAA liver injury model

[0175]

[0176] According to the experimental results in Table 2, the AST and ALT levels in the treatment group were significantly lower than those in the control group, indicating that the fermented Chinese herbal composition provided in the present application can effectively inhibit and reduce the experimental liver damage caused by CCl4 and TAA, protect the liver's free radical scavenging system, reduce AST and ALT values, and thus alleviate liver damage, indicating that the fermented Chinese herbal composition provided in the present application has a good effect in protecting the liver.

[0177] The data from the treatment groups of Examples 1-3 show that in Example 2, the mass ratio of the Chinese herbal composition to the probiotic composition is 2000:20, which is a relatively moderate ratio, ensuring both an adequate supply of Chinese herbal medicines and a sufficient number of probiotics to support an effective fermentation process, thereby achieving a better therapeutic effect.

[0178] Example 6 Clinical efficacy observation test

[0179] (1) 120 patients who met the physical examination criteria in the outpatient department were randomly selected. The treatment group included 80 cases, including 40 males and 40 females, aged 26-69 years, with a medical history of 60 days to 20 years. The control group included 30 cases, including 15 males and 15 females, aged 24-71 years, with a medical history of 60 days to 25 years. There was no statistically significant difference in the general data such as gender, age, and course of disease before treatment between the two groups.

[0180] (2) The inclusion criteria refer to the diagnostic criteria for non-alcoholic fatty liver disease developed by the Fatty Liver and Alcoholic Liver Disease Group of the Hepatology Branch of the Chinese Medical Association; patients who meet any of the following items 1-5 and 6 can be included.

[0181] 1) No history of drinking or alcohol consumption equivalent to less than 140g per week for men and less than 70g per week for women;

[0182] 2) Excluding specific diseases that can cause fatty liver, such as viral hepatitis, drug-induced fatty liver, total parenteral nutrition, and hepatolenticular degeneration;

[0183] 3) In addition to the clinical manifestations of the primary disease, there may be non-specific symptoms and signs such as fatigue, indigestion, dull pain in the liver area, and hepatosplenomegaly;

[0184] 4) May have metabolic syndrome-related components such as overweight and / or visceral obesity, increased fasting blood sugar, dyslipidemia, and hypertension;

[0185] 5) Serum transaminase and γ-glutamyl transpeptidase levels may be slightly or moderately elevated, usually with alanine aminotransferase (ALT) being the main elevated level;

[0186] 6) Liver imaging findings meet the imaging diagnostic criteria for diffuse fatty liver.

[0187] (3) Exclusion criteria: patients with hyperlipidemia caused by drugs; patients taking heparin, thyroxine, or other drugs that affect metabolism, and patients who have used other lipid-lowering methods in the past two weeks; patients with serious primary diseases such as cardiovascular, liver, kidney, and hematopoietic system diseases, and mental illness; and pregnant and lactating women.

[0188] (4) Treatment methods

[0189] The patients in the treatment group took the fermented Chinese herbal medicine composition prepared in Example 2 of the present application (the dosage form was selected according to the patient's condition) three times a day, with warm water after meals. During the treatment period, any lipid-lowering drugs were discontinued, the diet was properly controlled, and more aerobic exercise was performed. After 8 weeks of treatment, blood lipids and liver B-ultrasound were tested.

[0190] Control group: Oral lipid-lowering drug atorvastatin, 1 tablet (40 mg) every night before bedtime, blood lipids were rechecked after 8 weeks of treatment.

[0191] (5) Treatment outcomes

[0192] 1) The efficacy criteria refer to the "Guidelines for Clinical Research on New Chinese Medicines for the Treatment of Hyperlipidemia" issued by the Ministry of Health. TCM efficacy criteria:

[0193] Recovery: Clinical symptoms and signs have improved, and the syndrome score has decreased by ≥95%.

[0194] Markedly effective: Clinical symptoms and signs improved, and the syndrome score decreased by ≥70%.

[0195] Effective: Clinical symptoms and signs improved, and syndrome scores decreased by ≥30%.

[0196] Ineffective: There is no significant improvement or aggravation of clinical symptoms and signs, and the syndrome score decreases by less than 30%.

[0197] Fatty liver is divided into mild, moderate and severe according to B-ultrasound images. Mild: the echo of the front half of the liver section is slightly thickened and enhanced, the echo of the back half of the liver section is slightly reduced, and the light band on the liver surface exists; moderate: the echo of the front half of the liver section is thick and strong, the echo of the back half of the liver section is reduced, the light band on the liver surface is improved, and far-field enhancement may appear; severe: the echo of the front half of the liver section is significantly thickened and enhanced, the echo of the back half of the liver section disappears, and the light band on the liver surface does not appear. After treatment, recovery is considered as recovery, improvement of 1-2 levels is considered as reduction, and no change or aggravation is considered as ineffective.

[0198] 2) Comparison of the changes of the results of the detection indicators before and after treatment: After treatment, the serum total cholesterol, triglycerides, low-density lipoprotein cholesterol, ALT, and aspartate aminotransferase (AST) values ​​were significantly decreased compared with those before treatment, and the high-density lipoprotein cholesterol value was significantly increased (P < 0.05). In the treatment group, 39 cases of liver ultrasound and biochemistry returned to normal, 32 cases were relieved, 9 cases were ineffective, and the total effective rate was 88.75%. In the control group, 10 cases were normal, 16 cases were relieved, 4 cases were ineffective, and the total effective rate was 86.67%. The difference between the two groups was statistically significant (P < 0.05).

[0199] In summary, the fermented Chinese herbal composition prepared in Example 2 can significantly improve blood lipid levels. After treatment, the subjects' serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol (LDL-C) decreased significantly, and high-density lipoprotein cholesterol (HDL-C) increased significantly, which helps to improve blood lipid metabolism, thereby reducing the burden on the liver, improving liver function, reducing oxidative stress, promoting lipid metabolism, and reducing inflammatory response, thereby achieving the effect of protecting and benefiting the liver. After the subjects used the fermented Chinese herbal composition of the present application, the ALT and AST levels decreased significantly, indicating that liver function was improved, and the liver of most patients returned to normal or was alleviated. The fermented Chinese herbal medicine combination provided in the present application can effectively improve the patient's liver function, has the effect of protecting and benefiting the liver, and has good prospects in clinical applications.

[0200] Although the present invention has been described in detail above by general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A liver-benefiting fermented Chinese herbal medicine composition, characterized in that: It is composed of a Chinese herbal medicine composition and a probiotic composition, wherein the mass ratio of the Chinese herbal medicine composition to the probiotic composition is (1500-2500): (10-30); The Chinese herbal medicine composition is composed of the following components in parts by weight: 20-60 parts of Artemisia capillaris, 50-80 parts of Astragalus, 10-20 parts of green tea extract, 10-30 parts of Plantago, 10-30 parts of Peppermint, 6-20 parts of Coix seeds, 5-18 parts of Carthamus tinctorius, 5-15 parts of Borneolum, 10-20 parts of Lycium barbarum, 10-20 parts of Lysimachia chinensis, 10-20 parts of Honeysuckle and 10-20 parts of Pumpkin; The probiotic composition is composed of the following components in parts by weight: 1-5 parts of Bacillus amyloliquefaciens, 1-5 parts of Bacillus subtilis, 0.5-3 parts of Lactobacillus plantarum, 1.5-4.5 parts of Bifidobacterium, 0.5-3 parts of pectinase and 0.2-2 parts of β-glucanase; The name of the Bacillus amyloliquefaciens is HMJZ-B1003A, the classification name is Bacillus amyloliquefaciens, the preservation number is CCTCC NO: M 20191088, the preservation date is December 23, 2019, the preservation unit is China Center for Type Culture Collection, and the preservation address is Wuhan University, Wuhan, China; the name of the Bacillus subtilis is HMJZ-B1005S, the classification name is Bacillus subtilis, the preservation number is CCTCC NO: M 2020237, the preservation date is June 28, 2020, the preservation unit is China Center for Type Culture Collection, and the preservation address is Wuhan University, Wuhan, China.

2. A method for preparing a liver-benefiting fermented Chinese herbal medicine composition, used for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 1, characterized in that: The following steps are involved: S1. Preparation of primary extract: According to the formula of the herbal composition, weigh each component and prepare a primary extract; S2. Preparing a secondary extract: heating the primary extract to enzymatically hydrolyze the primary extract to prepare a secondary extract; S3. Preparation of probiotic fermentation broth: Bacillus amyloliquefaciens, Bacillus subtilis, Lactobacillus plantarum and Bifidobacterium were fermented and mixed to obtain a probiotic fermentation broth; S4 inoculation and multi-stage fermentation: The secondary extract was inoculated with the probiotic composition fermentation broth obtained in S3 and cultured and fermented to obtain a fermentation broth; S5. Inactivation and sealed anaerobic fermentation.

3. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 2, characterized in that: The S1 comprises the following specific steps: The Chinese medicine composition is weighed, mixed and crushed according to the formula, and then 1-2 times the volume of water is added to the Chinese medicine composition for soaking for 6-10 hours. The soaked Chinese medicine composition is extracted through an extraction device to obtain an extract, and the extract is sterilized by ultrafiltration.

4. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 2, characterized in that: The S2 comprises the following specific steps: Heat the mixed medicinal material extract, then add pectinase and heat to 35-60℃ for enzymatic hydrolysis for 15-30min. Use iodine solution for detection. When the medicinal solution does not turn blue when exposed to iodine solution, the enzymatic hydrolysis is completed. Then heat to 100℃ again to inactivate the enzyme for 10-20min. Add impurity remover, stir evenly, let stand for 2h and filter to obtain the secondary extract for use.

5. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 2, characterized in that: The S4 comprises the following specific steps: When the temperature of the secondary extract drops to 25-35°C, the probiotic composition fermentation liquid is inoculated and cultured until the pH value drops to 5.5-6.5, glucose is added and sterilized at high temperature, and when the temperature drops to 35-40°C, the fermentation is continued for 20-24 hours. When the pH value drops to 4.1-4.5, the fermentation temperature is lowered to 15-25°C, and the fermentation is continued until the pH value drops to 3.5-3.8, and the fermentation is terminated.

6. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 2, characterized in that: The S5 comprises the following specific steps: adding β-glucanase to the fermentation broth and mixing evenly, maintaining the broth at a constant temperature of 30-60° C. for 24-48 hours, and performing sealed anaerobic fermentation for 36-72 hours to obtain the final fermented Chinese herbal medicine composition.

7. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 4, characterized in that: The impurity remover in S2 is activated carbon, the addition amount is 0.5 g / L, and the addition amount of pectinase is 3 wt % of the medicinal material extract.

8. The method for preparing the liver-benefiting fermented Chinese herbal medicine composition according to claim 6, characterized in that: The amount of β-glucanase added in S5 is 5 wt% of the fermentation broth.

9. Use of the liver-benefiting fermented Chinese herbal medicine composition according to claim 1 in the preparation of liver-benefiting drugs.

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