Traditional Chinese medicine composition for protecting gastric mucosa and application

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for protecting gastric mucosa and application. The traditional Chinese medicine composition comprises the following raw materials: rhizoma corydalis and szechwan chinaberry fruit, and the preparation method comprises the following steps: adding water into the rhizoma corydalis, extracting, and filtering to obtain a rhizoma corydalis extracting solution and rhizoma corydalis dregs; then adding water into the dregs of the szechwan chinaberry fruit and the corydalis tuber, mixing, and adding enzyme for enzymolysis to obtain enzymatic hydrolysate; and finally, mixing the rhizoma corydalis extracting solution with the enzymatic hydrolysate, sterilizing, adding a probiotic liquid for fermentation, and filtering to obtain the rhizoma corydalis probiotic liquid. According to the composition prepared by the preparation method disclosed by the invention, through reasonable compatibility of the toosendan fruit and the rhizoma corydalis, the protective effect on gastric mucosa is remarkably superior to that of other methods, the lesion degree of the gastric mucosa of experimental gastric mucosal lesion model animals can be remarkably relieved, and the repairing capacity of the gastric mucosa is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine composition for protecting gastric mucosa and application thereof. Background Art

[0002] Gastric mucosa is an important part of the stomach wall and plays a vital role in protecting the stomach. However, due to factors such as irregular diet, high mental stress, Helicobacter pylori infection and long-term use of certain drugs, the incidence of gastric mucosal damage is increasing, which in turn causes a series of gastric diseases such as gastritis and gastric ulcer, seriously affecting the quality of life.

[0003] At present, there are many kinds of drugs on the market for protecting gastric mucosa, including chemical drugs such as bismuth and aluminum, as well as some traditional Chinese medicine preparations. However, chemical drugs often have side effects to varying degrees. For example, long-term use of bismuth may cause bismuth to accumulate in the body, causing adverse reactions such as the nervous system; aluminum may affect the intestinal absorption of phosphorus, etc.

[0004] Traditional Chinese medicine treatment has many advantages, but some products also have toxic ingredients, multiple medicinal flavors, complex ingredients that are difficult to control quality, and unclear efficacy advantages.

[0005] Toosendan is the dried mature fruit of Toosendan of Meliaceae, which has the effects of soothing liver and relieving heat, promoting qi and relieving pain, and killing insects. Corydalis is the dried tuber of Corydalis of Poppyaceae, which has the effects of promoting blood circulation, promoting qi and relieving pain. In traditional Chinese medicine theory, the two are often used to treat liver depression and qi stagnation, stomach pain and other symptoms. However, there are few in-depth studies on the combination of Toosendan and Corydalis in protecting gastric mucosa, and Toosendan contains toxic components such as azadirachtin. How to obtain a composition with stable quality, significant protection of gastric mucosal activity and removal of toxic and side effects through a specific preparation process has not been reported. Summary of the invention

[0006] In view of the problems existing in the prior art, the present invention provides a traditional Chinese medicine composition for protecting gastric mucosa and its application.

[0007] In order to achieve the purpose of the invention, the technical solution adopted is as follows:

[0008] A Chinese medicine composition for protecting gastric mucosa, characterized in that the preparation method of the Chinese medicine composition comprises the following steps:

[0009] (1) extracting Corydalis yanhusuo with water, filtering, and obtaining Corydalis yanhusuo extract and Corydalis yanhusuo residue;

[0010] (2) mixing the residues of Toosendan Fructus and Corydalis yanhusuo with water, and adding enzymes for enzymolysis to obtain enzymolysis solution;

[0011] (3) Mix the Corydalis yanhusuo extract and the enzymatic hydrolysate, sterilize, add probiotics solution for fermentation, and filter to obtain the product.

[0012] Preferably, the mass ratio of Corydalis yanhusuo to Toosendan fruit is 1:0.5-2, the mass ratio of Corydalis yanhusuo to water in step (1) is 1:5-12, the extraction is heating reflux extraction, the number of extractions is 1-3 times, and the time of each extraction is 0.5-1h.

[0013] Preferably, the enzymes in step (2) are pectinase, xylanase and tannase.

[0014] Preferably, the mass ratio of pectinase, xylanase and tannase is 1:0.2-0.5:0.8-1.5.

[0015] Preferably, the pH of the enzymatic hydrolysis in step (2) is 3.5-6.5, the enzymatic hydrolysis time is 3-5 hours, and the enzymatic hydrolysis temperature is 40-60°C.

[0016] Preferably, the mass of the water added in step (2) is 1-5 times the total mass of Toosendan Fructus and Corydalis yanhusuo residues, and the amount of enzyme added is 0.1-0.5% of the mass of the water added.

[0017] Preferably, the probiotic liquid in step (3) comprises inoculating Lactobacillus casei LC-12 into a seed culture medium and fermenting at 30-38° C. for 12-24 hours to obtain a probiotic liquid.

[0018] Preferably, the inoculation amount of the probiotic solution in step (3) is 0.5-1.5% of the total volume of the Corydalis extract and the enzymatic hydrolyzate.

[0019] Preferably, the fermentation temperature in step (3) is 26-30° C., and the fermentation time is 12-72 h.

[0020] Another object of the present invention is to provide the use of the above-mentioned traditional Chinese medicine composition in the preparation of a product for protecting gastric mucosa.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) The composition prepared by the preparation method of the present invention has a significantly better protective effect on gastric mucosa than other processes. The experimental results show that the composition can significantly reduce the degree of gastric mucosal lesions in experimental gastric mucosal injury model animals, improve the antioxidant capacity and repair capacity of gastric mucosa, and its efficacy is better than the commercially available Sanjiu Weitai group and first-line drugs such as cimetidine, omeprazole, etc.

[0023] (2) The preparation method of the present invention can effectively enrich active ingredients and remove impurities. Through strict selection of raw materials, optimization of extraction and fermentation processes, the content of azadirachtin is reduced, good safety data is accumulated, and the potential hepatotoxicity of the prescription is reduced.

[0024] (3) The composition of the present invention has high safety after preparation, provides reliable guarantee for clinical application, and has broad application prospects: Due to the high incidence of diseases related to gastric mucosal damage, the market demand for drugs to protect gastric mucosa is huge. The composition of the present invention has significant gastric mucosal protective activity, good quality stability and safety, and can be used to prepare drugs in various dosage forms, and new pH-dependent preparations can be given priority, with targeted release in the stomach to improve its bioavailability. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 These are example results of the gross morphological observation of gastric tissue in each group of rats.

[0026] Figure 2 This is a graph showing changes in the levels of inflammatory factors IL-6, TNF-α, PGE2, and CRP in rat serum. DETAILED DESCRIPTION

[0027] The following raw materials were all commercially available conventional materials, among which Lactobacillus casei LC-12 was purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd., and the supplier of pectinase, cellulase, xylanase, saccharifying enzyme and tannase was Nanning Dongheng Huadao Biotechnology Co., Ltd.

[0028] Example 1

[0029] A Chinese medicine composition for protecting gastric mucosa, the raw materials are Corydalis yanhusuo and Toosendan fruit in a mass ratio of 1:1, and the preparation method comprises the following steps:

[0030] (1) Add 8 times the amount of water to Corydalis yanhusuo and heat it under reflux for extraction. The extraction times are 2 times, and the extraction time for each time is 0.5 h. Filter and combine the filtrate to obtain Corydalis yanhusuo extract and Corydalis yanhusuo residue;

[0031] (2) Mix the residues of Toosendan Fructus and Corydalis yanhusuo with 5 times the amount of water, and add 0.5% enzyme by weight of water (mass ratio 1:0.2:

[0032] 1 of pectinase, xylanase and tannase), enzymolysis at pH 4.5 and 50°C for 4h to obtain enzymolysis solution;

[0033] (3) The Corydalis extract and the enzymatic hydrolysate are mixed, sterilized at 120° C. for 20 min, and a probiotic solution is added at 1% of the total volume of the Corydalis extract and the enzymatic hydrolysate, and fermented at 30° C. for 24 h, filtered, concentrated under reduced pressure at 65° C., and dried at 105° C.

[0034] The probiotic bacterial liquid is prepared by inoculating Lactobacillus casei LC-12 into a seed culture medium (composed of 1.0 g of glucose, 1.0 g of peptone, 0.5 g of yeast powder, 0.5 g of sodium chloride and 100 mL of pure water), and fermenting at 35° C. for 24 hours to obtain the probiotic bacterial liquid.

[0035] Example 2

[0036] A Chinese medicine composition for protecting gastric mucosa, the raw materials are Corydalis yanhusuo and Toosendan fruit in a mass ratio of 1:1, and the preparation method comprises the following steps:

[0037] (1) Add 5 times the amount of water to Corydalis yanhusuo and heat it under reflux for extraction. The number of extractions is 3 times, and the time of each extraction is 0.5 h. Filter and combine the filtrate to obtain Corydalis yanhusuo extract and Corydalis yanhusuo residue;

[0038] (2) Mix the residues of Toosendan and Corydalis with 1 times the amount of water, add a mixture of pectinase, xylanase and tannase in a mass ratio of 1:0.2:0.8 (0.5% by mass of water), and perform enzymolysis at pH 3.5 and 60°C for 3 hours to obtain an enzymolysis solution;

[0039] (3) Mix the Corydalis extract and the enzymatic hydrolysate, sterilize at 120° C. for 20 min, add 0.5% probiotic solution of the total volume of the Corydalis extract and the enzymatic hydrolysate, ferment at 26° C. for 12 h, filter, concentrate under reduced pressure at 65° C., and dry at 105° C.

[0040] The probiotic solution is prepared by inoculating Lactobacillus casei LC-12 into a seed culture medium (composed of 1.0 g glucose, 1.0 g peptone, 0.5 g yeast powder, 0.5 g sodium chloride and 100 mL pure water), and fermenting at 38° C. for 12 h to obtain the probiotic solution.

[0041] Example 3

[0042] A Chinese medicine composition for protecting gastric mucosa, the raw materials are Corydalis yanhusuo and Toosendan fruit in a mass ratio of 1:1, and the preparation method comprises the following steps:

[0043] (1) Add 12 times the amount of water to Corydalis yanhusuo and heat under reflux for extraction for 1 hour, filter, and obtain Corydalis yanhusuo extract and Corydalis yanhusuo residue;

[0044] (2) Mix the residues of Toosendan and Corydalis with twice the amount of water, add a mixture of pectinase, xylanase and tannase in a mass ratio of 1:0.5:1.5 (0.5% of the mass of water), and perform enzymolysis at pH 6.5 and 40° C. for 5 h to obtain an enzymolysis solution;

[0045] (3) Mix the Corydalis extract and the enzymatic hydrolysate, sterilize at 120° C. for 20 min, add 1.5% probiotic solution of the total volume of the Corydalis extract and the enzymatic hydrolysate, ferment at 26° C. for 12 h, filter, concentrate under reduced pressure at 65° C., and dry at 105° C. to obtain the product.

[0046] The probiotic liquid is prepared by inoculating Lactobacillus casei LC-12 into a seed culture medium (composed of 1.0 g glucose, 1.0 g peptone, 0.5 g yeast powder, 0.5 g sodium chloride, and 100 mL pure water), and fermenting at 30° C. for 12 h to obtain the probiotic liquid.

[0047] Example 4

[0048] The difference between this embodiment and embodiment 1 is that the probiotics are different, specifically Lactobacillus casei CGMCC 1.29. The bacterial solution is cultured by inoculating Lactobacillus casei CGMCC 1.29 into a seed culture medium (composed of 1.0 g glucose, 1.0 g peptone, 0.5 g yeast powder, 0.5 g sodium chloride, and 100 mL pure water), and fermenting at 28° C. for 24 h to obtain a probiotic bacterial solution.

[0049] Example 5

[0050] The difference between this embodiment and embodiment 1 is that the enzymes are different, specifically a mixture of pectinase, cellulase and saccharifying enzyme in a ratio of 1:0.2:1 is used.

[0051] Example 6

[0052] The difference between this embodiment and embodiment 1 is that there is no fermentation, specifically:

[0053] A Chinese medicine composition for protecting gastric mucosa, the raw materials are Corydalis yanhusuo and Toosendan fruit in a mass ratio of 1:1, and the preparation method comprises the following steps:

[0054] (1) Add 8 times the amount of water to Corydalis yanhusuo and heat under reflux for extraction. The extraction times are 2 times, and the extraction time for each time is 0.5 h. Filter and combine the filtrates to obtain Corydalis yanhusuo extract and Corydalis yanhusuo residue;

[0055] (2) Mix the residues of Toosendan Fructus and Corydalis yanhusuo with 5 times the amount of water, and add 0.5% enzyme by weight of water (mass ratio 1:0.2:

[0056] 1 pectinase, xylanase and tannase), enzymolysis at pH 4.5 and 50°C for 4 hours to obtain enzymolysis solution;

[0057] (3) The Corydalis extract and the enzymatic hydrolyzate are mixed, concentrated under reduced pressure at 65°C, and dried at 105°C.

[0058] Example 7

[0059] The difference between this embodiment and embodiment 1 is that the probiotics are different, specifically Lactobacillus plantarum JYLP-002 (purchased from Shandong Zhongke Jiayi). The bacterial liquid is cultured by inoculating Lactobacillus plantarum JYLP-002 into a seed culture medium (composed of 1.0 g glucose, 1.0 g peptone, 0.5 g yeast powder, 0.5 g sodium chloride, and 100 mL pure water), and fermenting at 37° C. for 24 h to obtain a probiotic bacterial liquid.

[0060] Drug efficacy experiments, animal drug efficacy experiments on the treatment of gastric mucosal damage

[0061] 1.1 Experimental animals

[0062] Male SD rats (200 ± 20 g), 36-56 days old. Purchased from Zhuhai Baishitong Biotechnology Co., Ltd., experimental unit use license number: SYXK (Guangdong) 2022-0125, license number: SCXK (Guangdong) 2020-0051. Ethics number: gdpulacspf2022552. SD rats were housed in the Experimental Animal Center of Guangdong Pharmaceutical University under natural light, temperature of 25 °C, and relative humidity of 65-70%. Free drinking water. All operations during the experiment complied with the experimental ethics principles of the Experimental Animal Center of Guangdong Pharmaceutical University.

[0063] 1.2 Drugs and reagents

[0064] Samples prepared in Examples 1-7, cimetidine (batch number: C13095070, Shanghai McLean Biochemical Technology Co., Ltd.), Sanjiu Weitai Granules (batch number 2304031F, China Resources Sanjiu Pharmaceutical Co., Ltd.), ultrapure water (self-made in the laboratory), rat interleukin-6 (IL-6) ELISA detection kit, rat tumor necrosis factor α (TNF-α) ELISA detection kit, rat prostaglandin E2 (PGE2) ELISA detection kit, rat C-reactive protein (CRP) ELISA detection kit, rat superoxide dismutase (SOD) ELISA detection kit, and rat malondialdehyde (MDA) ELISA detection kit.

[0065] 1.3 Animal grouping and drug administration

[0066] Healthy SPF male SD rats, 36-56 days old (200 ± 20g), free diet, adaptive feeding for three days, were randomly divided into 11 groups according to body weight, namely control group (CON), model group (MOD), Sanjiu Weitai group (SJWT), cimetidine group (CTD), and Example 1-7 group, 10 in each group. The control group and the model group were given appropriate amount of distilled water by gavage; Sanjiu Weitai group was given Sanjiu Weitai liquid by gavage at 4.2g / kg / d; Cimetidine group was given cimetidine liquid by gavage at 42mg / kg / d; Example 1-7 group was given liquid prepared by 0.375g crude drug amount / kg / d corresponding group. The drug was administered at a fixed time every day, and each group was continuously administered for 1 week. Rats were fasted and water was not allowed 24h before the last administration; 1.5h after the last administration, except for the control group, the remaining rats were given 1mL of anhydrous ethanol, and the rats in the control group were given distilled water by gavage. One hour later, the SD rats were anesthetized by inhalation of isoflurane and samples were collected.

[0067] 1.4 Detection of gastric tissue biochemical indicators

[0068] Cut the stomach tissue of about 1cm*1cm in size at the obvious damaged part, put it in a cryopreservation tube, and store it in a -80℃ refrigerator. The biochemical indicators of stomach tissue include malondialdehyde (MDA) and superoxide dismutase (SOD). The operation was carried out according to the instructions of the kit.

[0069] 1.5 Gross observation and scoring of gastric tissue morphology

[0070] After the experimental rats were anesthetized by inhalation of isoflurane, the whole stomach was removed, the pylorus was ligated, and an appropriate amount of 10% paraformaldehyde solution was perfused. After fixation for 20 minutes, the stomach was cut open at the greater curvature, rinsed with PBS, and placed under a stereoscope for observation and photography to record the damage of the gastric mucosa. At the same time, the damage of the gastric mucosa was observed, the bleeding points and bleeding cords were counted, and the length and width of the bleeding cords were measured with a ruler. The degree of gastric injury of the rats in each group was scored according to the standards in Table 1 and Table 2.

[0071] Table 1 Gastric injury degree scoring table

[0072]

[0073] Table 2 Scoring criteria for bleeding points and bleeding lines in gastric injury

[0074]

[0075] Observation indicators: The degree of gastric mucosal injury in rats of each experimental group was expressed by injury incidence, injury integral index and injury inhibition rate.

[0076] Total score = bleeding point score + length score + width score × 2;

[0077] Injury incidence (%) = number of rats with bleeding or ulcers / number of rats in the group × 100;

[0078] Injury score index = total injury score of each group / number of animals in each group;

[0079] Damage inhibition rate (%) = (AB) / A×100;

[0080] Wherein, A-damage score of model group; B-damage score of dose group.

[0081] 2 Statistical methods

[0082] The experimental data were statistically analyzed using Graph Pad Prism 9.0.0, and the experimental results were expressed as mean ± standard deviation (x ± s). The data between the two groups were compared using the t test, and the mean values ​​between multiple groups were compared using one-way analysis of variance (One-Way ANOVA). P < 0.05 indicated that the difference was statistically significant, and P < 0.01 indicated that the difference was statistically significant.

[0083] 3 Experimental results and analysis

[0084] 3.1 Changes in SOD activity and MDA content of antioxidant-related factors in gastric tissue of rats in each group

[0085] The results showed that compared with the control group, the SOD level of the gastric tissue of the rats in the model group was significantly reduced (P < 0.01), the MDA level was significantly increased (P < 0.01), and the SOD / MDA ratio was significantly reduced (P < 0.01); compared with the model group, the SOD level of each drug administration group was significantly increased (P < 0.01), and the MDA level was significantly reduced (P < 0.01), and it was dose-related. Only the SOD and MDA levels of the Example 1-3 groups were close to the positive drug, especially the Example 1 group, and there was no statistical difference. The results are shown in Table 3.

[0086] Table 3

[0087]

[0088]

[0089] Note: Compared with the control group, the model group *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01; compared with the Example 1 group, &P<0.05, &&P<0.01.

[0090] 3.2 Gross observation of gastric tissue

[0091] The results showed that the gastric mucosa of the rats in the control group was smooth, without congestion, edema, bleeding or erosion; the gastric mucosa of the model group showed punctate or stripe-like bleeding, and large-area erosion. Compared with the model group, punctate or stripe-like bleeding was still observed in each drug-treated group, but the degree of gastric damage was improved. There was only a small amount of bleeding in the Sanjiu Weitai group and the cimetidine group, and the gastric surface was smoother than the model group, without erosion; there was improvement in Example 6; there was little bleeding in Examples 4-5 and 7, and the inflammation was reduced, with significant improvement; the protective effect of Example 1-3 on the gastric mucosa was greatly increased, with only tiny bleeding spots and no diffuse inflammatory changes in the entire stomach. The results are shown below Figure 1 .

[0092] 3.3 Results of the study on gastric injury incidence, injury score index, and injury inhibition rate

[0093] The results are shown in Table 4 below.

[0094] Table 4

[0095]

[0096]

[0097] Note: Compared with the control group, the model group *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01; compared with the Example 1 group, &P<0.05, &&P<0.01.

[0098] The results showed that compared with the control group, the gastric mucosa of the rats in the model group was damaged; compared with the model group, the incidence of gastritis in the positive drug group (CTD, SJWT) and Examples 1-7 was reduced. Among them, Example 1-3 had the best effect. Compared with the model group, the total score of gastric injury in the positive drug group (CTD, SJWT) and the total score of gastric injury in the Example 1-7 group was significantly reduced.

[0099] The present invention further studies and verifies the contents of IL-6, TNF-α, PGE2 and CRP in the serum of rats in the preferred embodiments 1-3. The results show that compared with the control group, the corresponding indicators of the rats in the model group were significantly increased (P < 0.01). Compared with the model group, the contents of IL-6, TNF-α, PGE2 and CRP in the serum of rats in the embodiment 1-3 groups were significantly reduced (P < 0.01). The specific results are shown in Figure 2 .

[0100] Summary: In the early stage of this project, the content of toosendanin was determined by the Toosendan detection method of the Chinese Pharmacopoeia. The results showed that the content of toosendanin in the composition prepared by the method of the present invention was lower than that of the composition prepared by other processes, indicating that the extraction and fermentation process of the present invention transformed the toosendanin and reduced the content of toosendanin. Combined with the above-mentioned efficacy experimental study of the present invention, the extract obtained effectively improved the efficacy of protecting the gastric mucosa, indicating that more effective active components were enriched or the stimulation to the gastric mucosa was reduced, and gastric damage was effectively inhibited. The specific efficacy material basis needs to be further confirmed.

Claims

1. A Chinese medicine composition for protecting gastric mucosa, characterized in that: The raw materials of the Chinese medicine composition are Corydalis yanhusuo and Toosendan fruit, and the preparation method comprises the following steps: (1) extracting Corydalis yanhusuo with water, filtering, and obtaining Corydalis yanhusuo extract and Corydalis yanhusuo residue; (2) mixing the residues of Toosendan Fructus and Corydalis yanhusuo with water, and adding enzymes for enzymolysis to obtain enzymolysis solution; (3) Mix the Corydalis yanhusuo extract and the enzymatic hydrolysate, sterilize, add probiotics solution for fermentation, and filter to obtain the product.

2. The Chinese medicine composition according to claim 1, characterized in that: The mass ratio of Corydalis yanhusuo to Toosendan fruit is 1:0.5-2, the mass ratio of Corydalis yanhusuo to water in step (1) is 1:5-12, the extraction is heating reflux extraction, the number of extractions is 1-3 times, and the time of each extraction is 0.5-1h.

3. The Chinese medicine composition according to claim 1, characterized in that The enzymes in step (2) are pectinase, xylanase and tannase.

4. The Chinese medicine composition according to claim 3, characterized in that: The mass ratio of pectinase, xylanase and tannase is 1:0.2-0.5:0.8-1.

5.

5. The Chinese medicine composition according to claim 1, characterized in that: The pH of the enzymatic hydrolysis in step (2) is 3.5-6.5, the enzymatic hydrolysis time is 3-5 hours, and the enzymatic hydrolysis temperature is 40-60°C.

6. The Chinese medicine composition according to claim 1, characterized in that: The mass of the water added in step (2) is 1-5 times the total mass of the residues of Toosendan Fructus and Corydalis yanhusuo, and the amount of the enzyme added is 0.1-0.5% of the mass of the water added.

7. The Chinese medicine composition according to claim 1, characterized in that: The probiotic liquid in step (3) comprises inoculating Lactobacillus casei LC-12 into a seed culture medium and fermenting at 30-38° C. for 12-24 hours to obtain a probiotic liquid.

8. The Chinese medicine composition according to claim 7, characterized in that: The inoculation amount of the probiotic solution in step (3) is 0.5-1.5% of the total volume of the Corydalis extract and the enzymatic hydrolyzate.

9. The Chinese medicine composition according to claim 7, characterized in that: The fermentation temperature in step (3) is 26-30° C., and the fermentation time is 12-72 hours.

10. Use of the traditional Chinese medicine composition according to any one of claims 1 to 9 in preparing a product for protecting gastric mucosa.

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