Traditional Chinese medicine composition for treating hepatic diseases as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention belongs to the technical field of traditional Chinese medicines, mainly relates to a traditional Chinese medicine composition and a preparation method thereof, and in particular relates to a traditional Chinese medicine composition for treating liver diseases as well as a preparation method and application thereof. According to the traditional Chinese medicine composition, oldenlandia diffusa, schisandra chinensis, radix pseudostellariae, lucid ganoderma and liquorice are selected, the performance is improved through staged fermentation of lactobacillus rhamnosus CICC-6133 and bifidobacterium breve CICC-6182, then water extraction, concentration and drying are conducted, and the traditional Chinese medicine composition is prepared, and the prepared traditional Chinese medicine composition has the improvement effect on alcoholic fatty liver, hepatitis B, gastric mucosal injury, cirrhosis ascites and advanced liver cancer.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine, and mainly relates to a traditional Chinese medicine composition and a preparation method thereof, in particular to a traditional Chinese medicine composition for treating liver diseases and a preparation method and application thereof. BACKGROUND

[0002] With the increasing concern of people on health, the demand for the treatment of liver diseases is also increasing. At present, the clinical treatment of liver diseases on the market mainly relies on chemical drugs. However, chemical synthetic drugs can inhibit inflammation or virus replication in the short term, but they have the problem of causing drug resistance, and long-term use may cause liver and kidney toxicity. In addition, the existing drugs on the market focus on a single pathogenesis, and have single efficacy, lacking of synergistic intervention on other co-morbidities. In view of these problems, the present application develops a traditional Chinese medicine composition, which selects medicinal materials with the same effect as food to improve alcoholic fatty liver, hepatitis B and gastric mucosa damage. SUMMARY

[0003] Therefore, the present application selects Sarcopyramis glabra, Schisandra chinensis, Pseudostellaria heterophylla, Ganoderma lucidum, Glycyrrhiza uralensis, Crataegus pinnatifida, Lycium barbarum and Lilium brownii to improve the performance by segmental fermentation with Lactobacillus rhamnosus CICC-6133 and Bifidobacterium breve CICC-6182, and the prepared traditional Chinese medicine composition has the effect of improving alcoholic fatty liver, hepatitis B and gastric mucosa damage.

[0004] Through research and development, the research team found a traditional Chinese medicine composition formula, the formula is Sarcopyramis glabra 4.2-6g, Schisandra chinensis 2.8-4g, Pseudostellaria heterophylla 1.4-2g, Ganoderma lucidum 1.4-2g, Glycyrrhiza uralensis 0.7-1; and found that it can be used for treating at least one of alcoholic fatty liver, cirrhosis ascites, hepatitis B, and advanced liver cancer; but the drug utilization of this formula is not high enough, in order to improve its effect, the researchers carried out biological fermentation on the formula ingredients, and further adjusted the formula: added several raw materials in the original formula, which are Crataegus pinnatifida 4.2-6g, Lycium barbarum 2.8-4g and Lilium brownii 2.8-4g; at the same time, the fermentation strains were screened, and it was found that Lactobacillus rhamnosus CICC-6133 and Bifidobacterium breve CICC-6182 fermentation can significantly improve alcoholic fatty liver, hepatitis B, and also has the effect of improving gastric mucosa damage.

[0005] In the first aspect, the present application provides a traditional Chinese medicine composition, which comprises Sarcopyramis glabra 4.2-6 parts by weight, Schisandra chinensis 2.8-4 parts by weight, Pseudostellaria heterophylla 1.4-2 parts by weight, Ganoderma lucidum 1.4-2 parts by weight, and Glycyrrhiza uralensis 0.7-1 part by weight.

[0006] Preferably, the traditional Chinese medicine composition is prepared by combined fermentation and extraction treatment of Lactobacillus rhamnosus CICC-6133 and Bifidobacterium breve CICC-6182.

[0007] Preferably, the traditional Chinese medicine composition further comprises hawthorn 4.2-6 parts, medlar 2.8-4 parts, and lily 2.8-4 parts.

[0008] Preferably, the proportions of the white flower snake tongue grass, schisandra chinensis, radix pseudostellariae, ganoderma lucidum, licorice, hawthorn, medlar, and lily are 3:2:1:1:0.5:3:2:2.

[0009] In a second aspect, the present application provides a preparation method of a traditional Chinese medicine composition.

[0010] S1, the traditional Chinese medicinal materials in the traditional Chinese medicine composition are weighed according to the prescription, and the traditional Chinese medicinal materials are cleaned, dried, and crushed for subsequent use;

[0011] S2, Lactobacillus rhamnosus CICC-6133 is inoculated on MRS agar medium and cultured at 30°C for 15-17h to activate the strain. The activated Lactobacillus rhamnosus is transferred to a 500-700mL MRS liquid culture medium shake flask, and then fermented for 8-10h to obtain a bacterial solution of 2.3-2.5×10 8 cfu / mL;

[0012] S3, Bifidobacterium breve CICC-6182 is inoculated on glucose blood liver agar medium and cultured at 37°C for 15-17h to activate the strain. The activated Bifidobacterium breve is transferred to a 500-700mL glucose blood liver liquid culture medium shake flask, and then fermented for 8-10h to obtain a bacterial solution of 1.8-2.0×10 8 cfu / mL;

[0013] S4, after the traditional Chinese medicinal materials prepared in step S1 are mixed uniformly, Lactobacillus rhamnosus CICC-6133 bacterial solution prepared in step S2 is added at an inoculation amount of 1-3% (v / w), and aerobic fermentation is carried out at 30°C for 18-24h to obtain a first product; then Bifidobacterium breve CICC-6182 bacterial solution prepared in step S3 is added at an inoculation amount of 1-3% (v / w), and anaerobic fermentation is carried out at 37°C for 18-24h. After fermentation, 3% H2O2 solution is used to inactivate Lactobacillus rhamnosus and Bifidobacterium breve in the fermentation product, and the desired traditional Chinese medicine fermentation product is obtained. The inoculation amount is calculated based on the total mass of the traditional Chinese medicinal materials;

[0014] S5, water is added to the traditional Chinese medicine fermentation product prepared in step S4 at 5-10 times, and water extraction is carried out in a water bath for 1.5h. The extraction is carried out 3 times, the filtrate is combined and concentrated to a crude drug concentration of 1.2g / mL, pre-frozen at -40°C for 3-4h, sublimed and dried at a vacuum degree of 10-20Pa and a rate of 0.5-0.1°C / h, and dried at 35°C until the water content is ≤3%. The obtained freeze-dried product is crushed to prepare a traditional Chinese medicine composition.

[0015] Preferably, the raw materials in step S1 are respectively crushed into 80-100 mesh particles.

[0016] Preferably, the MRS agar medium used in step S2 comprises: casein peptone 10 g, beef infusion powder 10 g, yeast extract powder 5 g, glucose 20 g, Tween 80 1 g, sodium acetate 5 g, triammonium citrate 2 g, K2HPO4 2 g, MgSO4.7H2O 0.2 g, MnSO4.H2O 0.05 g, agar 15 g, and distilled water 1000 mL.

[0017] Preferably, the glucose blood agar medium used in step S3 comprises: beef infusion powder 3 g, protamine peptone 10 g, tryptone 5 g, vegetable peptone 3 g, yeast extract powder 5 g, liver extract 150 mL, glucose 10 g, soluble starch 0.5 g, solution A 10 mL (K2HPO4 10 g, KH2PO4 10 g, distilled water 100 mL), solution B 5 mL (MgSO4.7H2O 4 g, NaCl 0.2 g, FeSO4.7H2O 0.2 g, MnSO4.H2O 0.2 g, distilled water 100 mL), 5% L-cysteine hydrochloride solution 10 mL, Tween 80 1 g, agar 15 g, defibrillated sheep blood 50 mL, and distilled water 825 mL.

[0018] Preferably, the water extraction in step S5 is carried out at a water addition ratio of 7-9 times.

[0019] Preferably, the water extraction in step S5 is carried out at a temperature of 90-92°C.

[0020] Preferably, the traditional Chinese medicine composition prepared by the method can be made into various dosage forms such as granules, tablets and capsules.

[0021] In a third aspect, the traditional Chinese medicine composition prepared by the method has an improving effect on alcoholic fatty liver, hepatitis B and gastric mucosa damage.

[0022] In a fourth aspect, the traditional Chinese medicine composition can be applied to the preparation of drugs.

[0023] The present application has the following beneficial effects:

[0024] 1. The present application has an improving effect on alcoholic fatty liver by segmental fermentation of Oldenlandia diffusa and Radix Pseudostellariae under fermentation conditions.

[0025] 2. The present application has an improving effect on hepatitis B by the synergistic effect of Schisandra chinensis fermentation liquor, Ganoderma lucidum fermentation liquor and Glycyrrhiza uralensis fermentation liquor.

[0026] 3. The wolfberry fermentation liquid and the lily fermentation liquid in the present invention work synergistically to improve gastric mucosal damage.

[0027] 4. In this invention, Hedyotis diffusa alleviates liver fibrosis by inhibiting the TGF-β / Smad pathway, promoting diuresis and reducing bloating. The anti-inflammatory and antioxidant effects of Ganoderma lucidum protect liver cells and slow the progression of cirrhosis. The hepatoprotective components of Schisandra chinensis stabilize liver cell membranes and reduce damage. Hawthorn improves portal circulation, reduces portal hypertension, and reduces ascites formation. These multiple mechanisms work together to provide anti-inflammatory and anti-fibrotic effects, regulate water metabolism, and improve ascites in cirrhosis.

[0028] 5. The triterpenoids in the Ganoderma lucidum used in this invention can inhibit the proliferation of liver cancer cells while enhancing the body's immune surveillance of liver cancer cells, helping to clear lesions. Hedyotis diffusa can specifically induce apoptosis in liver cancer cells, reducing the tumor burden in advanced liver cancer. Lycium barbarum polysaccharides and betaine can regulate immune function and enhance the body's anti-tumor response to liver cancer. Pseudostellaria pseudoginseng replenishes Qi and strengthens the body's body, alleviating the toxic side effects of chemotherapy and radiotherapy in advanced liver cancer. These herbs work synergistically to effectively inhibit the progression of advanced liver cancer, improve patients' immune status, and enhance their quality of life. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0030] Figure 1 This is a comparison chart of ALT activity in the serum of each group of mice in Example 1.

[0031] Figure 2 This is a comparison chart of AST activity in the serum of each group of mice in Example 1.

[0032] Figure 3 This is a comparison chart of LDH activity in the serum of each group of mice in Example 1.

[0033] Figure 4 This is a comparison chart of the TG content in the serum of each group of mice in Example 1.

[0034] Figure 5 3 is a comparison chart of the number of gastric mucosal bleeding points in rats of each group in Example 3.

[0035] Figure 6 This is a sample diagram of the Chinese medicine composition prepared by the present invention.

[0036] Following specific examples illustrate the embodiments of the present application, and other advantages and effects of the present application can be easily understood by those skilled in the art from the disclosure of this specification. The present application can also be implemented or applied by other different specific embodiments, and various modifications or changes can be made to the details in this specification based on different views and applications without departing from the spirit of the present application. It should be noted that the following examples and features in the examples can be combined with each other without conflict.

[0037] Oldenlandia diffusa, Schisandra chinensis, Radix Pseudostellariae, Ganoderma lucidum, Glycyrrhiza uralensis, Crataegus pinnatifida, Lycium barbarum and Lilium brownii were purchased from Bozhou Fushuotang Biological Technology Co., Ltd.; Lactobacillus rhamnosus CICC-6133 and Bifidobacterium breve CICC-6182 were purchased from China General Microbiological Culture Collection Center.

[0038] Example 1

[0039] This example proves that the segmented fermentation of Oldenlandia diffusa and Radix Pseudostellariae under fermentation conditions has an improvement effect on alcoholic fatty liver.

[0040] This example provides a traditional Chinese medicine composition, and the specific steps are as follows:

[0041] S1, 4.2 g of Oldenlandia diffusa, 2.8 g of Schisandra chinensis, 1.4 g of Radix Pseudostellariae, 1.4 g of Ganoderma lucidum, and 0.7 g of Glycyrrhiza uralensis were weighed, then cleaned, dried in an oven, and crushed to 80 mesh particles for standby;

[0042] S2, Lactobacillus rhamnosus CICC-6133 was inoculated on MRS agar medium and cultured at 30°C for 15 h to activate the strain. The activated Lactobacillus rhamnosus was transferred to a 500 mL MRS liquid medium shake flask and fermented for 8 h, finally obtaining a bacterial liquid of 2.3 x 10 8 cfu / mL;

[0043] S3, Bifidobacterium breve CICC-6182 was inoculated on glucose blood liver agar medium and cultured at 37°C for 15 h to activate the strain. The activated Bifidobacterium breve was transferred to a 500 mL glucose blood liver liquid medium shake flask and fermented for 8 h, finally obtaining a bacterial liquid of 1.8 x 10 8 cfu / mL;

[0044] S4, inoculate the raw materials prepared in step S1 with Lactobacillus rhamnosus CICC-6133 bacterial solution prepared in step S2 at an inoculation amount of 3% (v / w) and perform aerobic fermentation at 30°C for 18h to obtain a first product; then inoculate the fermentation product with Bifidobacterium breve CICC-6182 bacterial solution prepared in step S3 at an inoculation amount of 1% (v / w) and perform anaerobic fermentation at 37°C for 18h; after the fermentation is completed, inactivate the Lactobacillus rhamnosus and Bifidobacterium breve in the fermentation product using 3% H2O2 solution to obtain the desired traditional Chinese medicine fermentation product; the inoculation amount is based on the total mass of the traditional Chinese medicinal materials;

[0045] S5, add 7 times the amount of water to the traditional Chinese medicine fermentation product prepared in step S4, stir until uniform, and then place in a water bath at 90°C for water extraction for 1.5h; repeat the extraction for 3 times, combine the filtrates after filtration, and concentrate to a crude drug concentration of 1.2g / mL; pre-freeze at -40°C for 3h, sublimate at a rate of 0.5°C / h under a vacuum degree of 10Pa, and dry at 35°C until the water content is ≤3%; after the obtained freeze-dried product is crushed, a traditional Chinese medicine composition is prepared.

[0046] Comparative Example 1: do not add Hydrangea patula in step S4, and the remaining steps are consistent with Example 1.

[0047] Comparative Example 2: do not add Radix Pseudostellariae in step S4, and the remaining steps are consistent with Example 1.

[0048] Comparative Example 3: do not add Lactobacillus rhamnosus CICC-6133 in step S4, and the remaining steps are consistent with Example 1.

[0049] Comparative Example 4: do not add Bifidobacterium breve CICC-6182 in step S4, and the remaining steps are consistent with Example 1.

[0050] Comparative Example 5: replace Lactobacillus rhamnosus CICC-6133 in step S4 with Lactobacillus rhamnosus CICC-21772 (purchased from China Industrial Microbial Culture Collection Center), and the remaining steps are consistent with Example 1.

[0051] Comparative Example 6: replace Bifidobacterium breve CICC-6182 in step S4 with Bifidobacterium breve CICC-24633 (purchased from China Industrial Microbial Culture Collection Center), and the remaining steps are consistent with Example 1.

[0052] Animals: The experimental animals selected were C57BL / 6J male mice, provided by Jiangsu Ailinfeng Biotechnology Co., Ltd., and the body weight was controlled at 20±2g. Before the formal start of the experiment, all mice needed to undergo a 7-day environmental adaptation period to ensure that they could adapt to the experimental environment. During the adaptation period, the mice were placed in cages with independent ventilation systems, and the temperature of the feeding environment was maintained at 25±2℃, the humidity was controlled at 50%-60%, and a 12h light-dark alternating photoperiod was provided. In addition, in order to adapt the mice to liquid diet, they were fed with maltodextrin liquid feed (maltodextrin and water 1:1) during this period.

[0053] Grouping: After 7d of adaptive feeding, 72 mice were randomly divided into 9 groups, namely the blank group, the model group, the example 1 group and the comparative examples 1-6 groups, 8 in each group.

[0054] Modeling: From the 8th day, except for the blank group, the rest of the mice were fed with maltodextrin liquid feed containing 5% alcohol, and the blank group was fed with an equal dose of maltodextrin liquid feed. At the same time, the mice in the example 1 group and the comparative examples 1-6 groups were given the corresponding traditional Chinese medicine composition by gavage at 8:30 am every day, 1.3g / kg; the blank group and the model group were given the same volume of normal saline. After 14d of continuous feeding, at 8:30 am the next day, except for the blank group, the other groups were given 100mg / 20g of maltodextrin containing 30% alcohol by gavage. 9h after the last gavage, the serum of the mice was collected for analysis.

[0055] Determination of serum indicators: blood samples were collected from the retro-orbital plexus of the mice. After blood collection, the samples were left to stand for 35min, and the serum was collected by centrifugation at 3500r / min for 15min at 4℃. The determination of ALT, AST, LDH activity and TG content in serum was carried out according to the instructions of the detection kit, and the experimental results are shown in Figure 1 、 Figure 2 、 Figure 3 and Figure 4 .

[0056] Figure 1 is a comparison chart of ALT activity in the serum of mice in each group; Figure 2 is a comparison chart of AST activity in the serum of mice in each group; Figure 3 is a comparison chart of LDH activity in the serum of mice in each group; Figure 4 is a comparison chart of TG content in the serum of mice in each group. From Figures 1-4It can be seen that compared with the blank group mice, the model group mice had significantly increased serum ALT, AST and LDH activities and TG content, indicating that alcohol diet caused liver damage in mice; compared with the model group mice, the serum ALT, AST and LDH activities and TG content of the mice in Example 1 group and Comparative Examples 1-6 group were significantly reduced, and the serum ALT, AST and LDH activities and TG content of the mice in Example 1 group were significantly lower than those in Comparative Examples 1-6 group, indicating that Example 1 group had the best improvement effect on alcohol diet-induced liver damage, confirming that the segmented fermentation of Oldenlandia diffusa and Radix Pseudostellariae under fermentation conditions had an improvement effect on alcoholic fatty liver.

[0057] Example 2

[0058] This example proves that the synergistic effect of Schisandra fermenting liquor, Ganoderma lucidum fermenting liquor and Glycyrrhiza uralensis fermenting liquor has an improvement effect on hepatitis B.

[0059] This example provides a traditional Chinese medicine composition, and the specific steps are as follows:

[0060] S1, weigh Oldenlandia diffusa 5.1 g, Schisandra chinensis 3.4 g, Radix Pseudostellariae 1.7 g, Ganoderma lucidum 1.7 g, Glycyrrhiza uralensis 0.85 g, Crataegus pinnatifida 5.1 g, Lycium barbarum 3.4 g and Lilium brownii 3.4 g, then clean them up, dry them in an oven, and crush them to 90 mesh particles for standby;

[0061] S2, inoculate Lactobacillus rhamnosus CICC-6133 on MRS agar medium, and culture at 30°C for 16 h to activate the strain. After activation, the Lactobacillus rhamnosus is transferred to a 600 mL MRS liquid medium shaking flask, and then fermented for 9 h to obtain a bacterial liquid of 2.4 x 10 8 cfu / mL;

[0062] S3, inoculate Bifidobacterium breve CICC-6182 on glucose blood liver agar medium, and culture at 37°C for 16 h to activate the strain. After activation, the Bifidobacterium breve is transferred to a 600 mL glucose blood liver liquid medium shaking flask, and then fermented for 9 h to obtain a bacterial liquid of 1.9 x 10 8 cfu / mL;

[0063] S4, inoculate the raw materials prepared in step S1 with Lactobacillus rhamnosus CICC-6133 bacterial solution prepared in step S2 at an inoculation amount of 2% (v / w), and perform aerobic fermentation at 30°C for 21h to obtain a first product; then inoculate the first product with Bifidobacterium breve CICC-6182 bacterial solution prepared in step S3 at an inoculation amount of 2% (v / w), and perform anaerobic fermentation at 37°C for 21h, after which inactivate the Lactobacillus rhamnosus and Bifidobacterium breve in the fermented product using 3% H2O2 solution to obtain the desired traditional Chinese medicine fermentation product; the inoculation amount is based on the total mass of the traditional Chinese medicinal materials;

[0064] S5, add 8 times the amount of water to the traditional Chinese medicine fermentation product prepared in step S4, stir until uniform, and then place in a water bath and extract in water at 91°C for 1.5h, repeat the extraction 3 times, combine the filtrates after filtration, and concentrate to a crude drug concentration of 1.2g / mL, pre-freeze at -40°C for 3.5h, sublimate at a rate of 0.8°C / h under a vacuum of 15Pa, and dry at 35°C until the water content is ≤3%, and then pulverize the obtained freeze-dried product to obtain a traditional Chinese medicine composition.

[0065] Comparative Example 7: In step S4, no Schisandra chinensis is added, and the remaining steps are consistent with Example 2.

[0066] Comparative Example 8: In step S4, no Ganoderma lucidum is added, and the remaining steps are consistent with Example 2.

[0067] Comparative Example 9: In step S4, no Glycyrrhiza uralensis is added, and the remaining steps are consistent with Example 2.

[0068] Animals: The experimental animals are SD rats provided by Jiangsu Ailinfeng Biological Technology Co., Ltd., with half male and half female, weighing 220±20g. All rats need to be adaptively fed for 7d before the experiment starts. The feeding system uses independent ventilation cages, and the environmental parameters are: temperature 25±2°C, humidity 50%-60%, and 12h light-dark alternating photoperiod.

[0069] Grouping: After adaptive feeding for 7d, the rats are randomly divided into a blank group, a model group, a positive control group, an Example 2 group, and Comparative Examples 7-9 groups, 8 rats in each group.

[0070] Modeling: Starting from the 8th day, rats in each group except the blank group are injected with HBV antigen (purchased from Shanghai Yushao Biological Technology Co., Ltd.) at a concentration of 1×10 6 / mL, 10mL / kg, once every 7d, for 21d, during which the rats are allowed to freely eat and drink. If the rats are HBV positive and have liver fibrosis and cirrhosis, it indicates that the modeling is successful. Rats in the blank group are injected with the same amount of normal saline in the tail vein.

[0071] Dosing: After the modeling is successful, the positive control group of rats is injected with 10 ug pKCMvint.IFN-α plasmid (purchased from Essen Hospital Virus Institute) through high-pressure tail vein, 7d / time, for 14d; the rats in Example 2 group and Comparative Examples 7-9 group are orally administered with the corresponding traditional Chinese medicine composition, the dosage is 0.9g / kg, 1d / time, for 14d; the rats in blank group and model group are orally administered with the same amount of normal saline, for 14d.

[0072] Determination: After the administration is completed, the rats are taken blood from the eye socket, centrifuged at 3500r / min for 15min to collect the serum, and the content of HBV markers (HBeAg and HBsAg) and liver function indicators (GOT and GPT) in the serum is determined according to the instruction of ELISA kit (purchased from Shenzhen Dot Biological Technology Co., Ltd.), and the experimental results are shown in Table 1. Figure 1

[0073] Table 1

[0074] Group HBeAg (U / mL) HBsAg (s / CD) GPT (U / L) GOT (U / L) Blank group — — 48.23±6.94 187.46±23.96 Model group 543.95±73.95 77.39±14.95 88.38±14.95 349.57±47.83 Example 2 group 295.72±35.50 44.48±8.27 65.39±8.84 281.48±28.01 Comparative example 7 group 508.60±53.13 69.24±11.68 79.79±6.48 321.68±31.77 Comparative example 8 group 484.82±32.58 61.33±8.16 76.48±6.94 314.12±24.79 Comparative example 9 group 450.35±28.48 57.95±12.46 71.65±6.58 336.97±35.55 Positive control group 184.23±24.74 25.45±5.35 61.75±9.01 272.46±25.12

[0075] Table 1 is a comparison table of the content of HBV markers (HBeAg and HBsAg) and liver function indicators (GOT and GPT) in the serum of rats in each group. As shown in Table 1, compared with the blank group of rats, the content of HBeAg, HBsAg, GOT and GPT in the serum of the model group of rats is significantly increased; compared with the model group of rats, the content of HBeAg, HBsAg, GOT and GPT in the serum of the rats in Example 2 group and Comparative Examples 7-9 group is significantly reduced, and the content of HBeAg, HBsAg, GOT and GPT in the serum of Example 2 group is significantly lower than that of the rats in Comparative Examples 7-9 group, which indicates that Example 2 group has the best improvement effect on hepatitis B, and confirms that the synergistic effect of Schisandra chinensis fermentation liquor, Ganoderma lucidum fermentation liquor and Glycyrrhiza uralensis fermentation liquor has an improvement effect on hepatitis B.

[0076] Example 3

[0077] This example proves that the synergistic effect of Lycium barbarum fermentation liquor and Lilium brownii fermentation liquor has an improvement effect on gastric mucosal injury.

[0078] This example provides a traditional Chinese medicine composition, and the specific steps are as follows:

[0079] S1, weigh 6g of Oldenlandia diffusa, 4g of Schisandra chinensis, 2g of Taizishen, 2g of Ganoderma lucidum, 1g of Glycyrrhiza uralensis, 6g of Crataegus pinnatifida, 4g of Lycium barbarum and 4g of Lilium brownii, then clean them up, dry them in an oven, and crush them to 100 mesh particles for standby;

[0080] ​S2, Lactobacillus rhamnosus CICC-6133 was inoculated on MRS agar medium and cultured at 30°C for 17 h to activate the strain. The activated Lactobacillus rhamnosus was transferred to a 700 mL MRS liquid medium shake flask and fermented for another 10 h to obtain a bacterial solution of 2.5 x 10 8 cfu / mL.

[0081] S3, Bifidobacterium breve CICC-6182 was inoculated on glucose blood liver agar medium and cultured at 37°C for 17 h to activate the strain. The activated Bifidobacterium breve was transferred to a 700 mL glucose blood liver liquid medium shake flask and fermented for another 10 h to obtain a bacterial solution of 2.0 x 10 8 cfu / mL.

[0082] S4, the raw materials prepared in step S1 were mixed uniformly, and then Lactobacillus rhamnosus CICC-6133 bacterial solution prepared in step S2 was added at an inoculation amount of 1% (v / w), and aerobic fermentation was carried out at 30°C for 24 h to obtain a first product; then Bifidobacterium breve CICC-6182 bacterial solution prepared in step S3 was added at an inoculation amount of 3% (v / w), and anaerobic fermentation was carried out at 37°C for 24 h. After fermentation, 3% H2O2 solution was used to inactivate Lactobacillus rhamnosus and Bifidobacterium breve in the fermentation product, thereby obtaining the desired traditional Chinese medicine fermentation product; the inoculation amount was calculated based on the total mass of the traditional Chinese medicinal materials;

[0083] S5, water was added to the traditional Chinese medicine fermentation product prepared in step S4 at a ratio of 9:1, and after stirring uniformly, the mixture was placed in a water bath at 92°C for water extraction for 1.5 h. The extraction was repeated for 3 times, and then the filtrates were combined and concentrated to a crude drug concentration of 1.2 g / mL. The mixture was pre-frozen at -40°C for 4 h, and then sublimation drying was carried out at a vacuum degree of 20 Pa and a rate of 0.1°C / h. The mixture was dried at 35°C until the water content was ≤3%. The obtained freeze-dried product was crushed to obtain a traditional Chinese medicine composition.

[0084] Comparative Example 10: In step S4, wolfberry was not added, and the remaining steps were consistent with Example 3.

[0085] Comparative Example 11: In step S4, lily was not added, and the remaining steps were consistent with Example 3.

[0086] Animals: SD rats provided by Jiangsu Ailinfeng Biological Technology Co., Ltd. were used as experimental animals, with half male and half female, and the body weight was 220 ± 20 g. All rats were adaptively fed for 7 days before the experiment started. The feeding system used independent ventilation cages, and the environmental parameters were: temperature 25 ± 2°C, humidity 50%-60%, and 12-hour light-dark alternating light cycle.

[0087] Grouping: After adaptive feeding for 7 days, the rats were randomly divided into 4 groups, namely the model group, the Example 3 group, the Comparative Examples 10-11 groups, 8 rats in each group.

[0088] Dosing and determination: The Example 3 group and the Comparative Examples 10-11 groups were orally administered with the corresponding traditional Chinese medicine composition, which was diluted with purified water to a concentration of 20% v / v before oral administration, and the oral administration dose was 0.9 g / kg; the model group was orally administered with the same amount of distilled water, once in the morning and once in the evening, continuously for 10 days. 1 hour after the last oral administration, the stomachs of all rats were ligated under sterile conditions to cause gastric mucosal injury and bleeding in rats, and the number of bleeding points in the gastric mucosa was observed under a microscope, and the experimental results are shown in Table 1. Figure 5

[0089] Figure 5 is a comparison chart of the number of bleeding points in the gastric mucosa of rats in each group. As can be seen directly from the chart, compared with the model group, the number of bleeding points in the gastric mucosa of rats in the Comparative Examples 10-11 groups had no significant difference, and the number of bleeding points in the gastric mucosa of rats in the Example 3 group was significantly reduced, indicating that the Example 3 group can significantly reduce the amount of bleeding in the gastric mucosa of rats, and the synergistic effect of the Gouqi fermentation liquor and the Baihe fermentation liquor has an improvement effect on the gastric mucosal injury of rats.

[0090] The above examples only express the specific embodiments of the present application, which are described in more detail and in more detail, but should not be understood as limiting the protection scope of the present application. It should be noted that for ordinary skilled persons in the art, any changes, modifications, substitutions, combinations, simplifications made without departing from the concept of the present application, without departing from the spirit and principles of the present application, should be equivalent to the replacement method, and all are included in the protection scope of the present application.​

Claims

1. A Chinese medicine composition for treating liver disease, characterized in that: The composition comprises 4.2-6 parts of Hedyotis diffusa herb, 2.8-4 parts of Schisandra chinensis chinensis, 1.4-2 parts of Pseudostellaria pseudoginseng, 1.4-2 parts of Ganoderma lucidum and 0.7-1 part of Licorice root by weight.

2. A traditional Chinese medicine composition for treating liver disease according to claim 1, wherein the traditional Chinese medicine composition is prepared by combined fermentation and extraction of Lactobacillus rhamnosus CICC-6133 and Bifidobacterium breve CICC-6182.

3. A traditional Chinese medicine composition for treating liver disease according to claim 2, further comprising 4.2-6 parts of hawthorn, 2.8-4 parts of wolfberry and 2.8-4 parts of lily.

4. A Chinese medicine composition for treating liver disease according to claim 3, characterized in that: The ratio of Hedyotis diffusa, Schisandra chinensis, Pseudostellaria pseudoginseng, Ganoderma lucidum, Licorice root, Hawthorn fruit, Lycium barbarum and Lilium is 3:2:1:1:0.5:3:2:

2.

5. A method for preparing a Chinese medicine composition for treating liver disease according to any one of claims 1 to 4, wherein the specific preparation method is as follows: S1. Weigh the Chinese medicinal materials in the Chinese medicine composition according to the formula, and wash, dry and crush the Chinese medicinal materials separately for use; S2. Lactobacillus rhamnosus CICC-6133 was inoculated onto MRS agar medium and cultured at 30°C for 15-17 hours to activate the strain. The activated Lactobacillus rhamnosus was transferred to a 500-700 mL MRS liquid medium shake flask and fermented for another 8-10 hours to obtain 2.3-2.5×10 8 cfu / mL bacterial solution for future use; S3. Bifidobacterium breve CICC-6182 was inoculated onto glucose-blood-liver agar medium and cultured at 37°C for 15-17 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500-700 mL glucose-blood-liver liquid medium shake flask and fermented for another 8-10 hours to obtain 1.8-2.0×10 8 cfu / mL bacterial solution for future use; S4, after uniformly mixing the Chinese medicinal material raw materials prepared in step S1, adding the Lactobacillus rhamnosus CICC-6133 bacterial solution prepared in step S2 at an inoculum amount of 1-3% (v / w), and performing aerobic fermentation at 30° C. for 18-24 hours to obtain a first product; then adding the Bifidobacterium breve CICC-6182 bacterial solution prepared in step S3 at an inoculum amount of 1-3% (v / w), and fermenting anaerobically at 37° C. for 18-24 hours. After the fermentation is completed, the Lactobacillus rhamnosus and Bifidobacterium breve in the fermentation product are inactivated with a 3% H2O2 solution to obtain the desired Chinese medicinal fermentation product, which is set aside; the inoculum amount is based on the total mass of the Chinese medicinal material raw materials; S5. Add 5-10 times of water to the fermented Chinese medicine prepared in step S4, extract in water in a water bath for 1.5 hours, extract three times, filter, combine the filtrate and concentrate to a crude drug concentration of 1.2 g / mL, pre-freeze at -40°C for 3-4 hours, sublimate and dry at a rate of 0.5-0.1°C / h under a vacuum degree of 10-20 Pa and dry at 35°C to a moisture content of ≤3%. The resulting freeze-dried product is crushed to prepare a Chinese medicine composition.

6. The method for preparing a Chinese medicine composition for treating liver disease according to claim 5, characterized in that: The components of the MRS agar medium used in step S2 are: 10 g casein peptone, 10 g beef extract powder, 5 g yeast extract powder, 20 g glucose, 1 g Tween 80, 5 g sodium acetate, 2 g triammonium citrate, 2 g K2HPO4, 0.2 g MgSO4·7H2O, 0.05 g MnSO4·H2O, 15 g agar, and 1000 mL distilled water.

7. The method for preparing a Chinese medicine composition for treating liver disease according to claim 5, characterized in that: The components of the glucose-blood-liver agar medium used in step S3 are: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g plant peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (10 g K2HPO4, 10 g KH2PO4, 100 mL distilled water), 5 mL solution B (4 g MgSO4·7H2O, 0.2 g NaCI, 0.2 g FeSO4·7H2O, 0.2 g MnSO4·H2O, 100 mL distilled water), 10 mL 5% L-cysteine ​​hydrochloride solution, 1 g Tween 80, 15 g agar, 50 mL defibrinated sheep blood, and 825 mL distilled water.

8. The method for preparing a Chinese medicine composition for treating liver disease according to claim 5, characterized in that: In step S5, the water addition ratio during water extraction is 7-9 times; The water extraction temperature in step S5 is 90-92°C.

9. The method for preparing a Chinese medicine composition for treating liver disease according to claim 5, characterized in that: The traditional Chinese medicine composition is prepared into granules, tablets and capsules.

10. The Chinese medicine composition according to claims 1 to 4 or the method for preparing a Chinese medicine composition for treating liver diseases according to claim 5 is used in the preparation of a drug for treating or improving alcoholic fatty liver, hepatitis B, and gastric mucosal damage.

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