Use of digestive aid tablets in preparation of drug for treating irritable bowel syndrome

Abstract

Use of digestive aid tablets in the preparation of a drug for treating irritable bowel syndrome. The digestive aid tablets consist of Citri Reticulatae Pericarpium, Pseudostellariae Radix, stir-fried Hordei Fructus Germinatus, Crataegi Fructus, and Dioscoreae Rhizoma, and exhibit significant efficacy in the treatment of irritable bowel syndrome with diarrhea and liver qi stagnation and spleen deficiency syndrome.

Description

Application of Jianwei Xiaoshi Tablets in preparation of a drug for treating irritable bowel syndrome TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine, and particularly relates to application of Jianwei Xiaoshi Tablets in preparation of a drug for treating irritable bowel syndrome. BACKGROUND

[0002] Irritable bowel syndrome (IBS) is a relatively common functional gastrointestinal disease, and the main clinical manifestations are recurrent abdominal pain, abdominal distension, bowel habits and changes in stool form. Due to individual differences, different symptoms are exhibited, and thus IBS can be divided into four subtypes, i.e., diarrhea-predominant IBS (IBS-D), constipation-predominant IBS (IBS-C), mixed-type IBS (IBS-M) and unsub-typed IBS (IBS-U). IBS is also considered as a typical psychosomatic disease, and has the pathogenic characteristics of multi-dimension and multi-target. The pathogenesis of IBS is complex and multi-dimensional, and is not formed by a single factor. Domestic and foreign researches show that the pathogenesis of IBS is mainly related to intestinal flora imbalance, gastrointestinal motility disorder, intestinal low-grade inflammation, visceral hypersensitivity, intestinal-brain interaction change, increased intestinal permeability, food intolerance, heredity and psychosocial factors. The disease has a long course and is difficult to cure, and seriously affects the quality of life of patients and their families, and brings intangible spiritual pressure to human beings. At present, there is still no unified standard for the treatment scheme of IBS with definite curative effect of western medicine, and the main treatment is symptomatic treatment, and the purpose of improving clinical symptoms, eliminating the concerns of patients and improving the quality of life can be achieved in the clinic.

[0003] There is no traditional medicine on irritable bowel syndrome, which is divided into "diarrhea" and "abdominal pain" and other categories according to clinical manifestations, and the disease is located in the intestine and involves the liver and spleen. According to the 2017 expert consensus on Chinese medicine diagnosis and treatment of irritable bowel syndrome, IBS-D is divided into liver stagnation and spleen deficiency, spleen deficiency and dampness, spleen and kidney yang deficiency, spleen and stomach dampness, and cold and heat mixed syndrome. Among them, the liver stagnation and spleen deficiency IBS-D has the highest prevalence. Liver stagnation and spleen deficiency IBS-D is mainly caused by emotional disorder, improper diet and invasion of external pathogens. The spleen is the basis of acquired constitution, and the spleen transports water and dampness, and is the source of blood and blood. Improper diet leads to insufficient source of blood and blood, and the spleen loses the function of transportation. At the same time, the liver and spleen are not adjusted, the liver loses the function of dredging, and the two are caused by this disease. In treatment, the main principle is to soothe the liver and invigorate the spleen, dry dampness and stop diarrhea. Good effect has been achieved in the treatment of IBS in clinic, and the method of soothing the liver and invigorating the spleen for treating liver stagnation and spleen deficiency IBS-D has become the main research type of traditional Chinese medicine. More people explore its pathogenesis, prevention and treatment methods from the perspective of traditional Chinese medicine, and provide new ideas and directions for clinical treatment and research.

[0004] Jianwei Xiaoshi Tablets are composed of five Chinese medicines, including Pericarpium Citri Reticulatae, Radix Pseudostellariae, Fried Hordeum Vulgare, Crataegus Pinnatifida and Dioscorea Opposita. All of them have the effect of invigorating the stomach and promoting digestion. Jianwei Xiaoshi Tablets, which are recorded in Chinese Pharmacopoeia 2010, have the effect of invigorating the spleen and promoting digestion, and are commonly used for treating indigestion in China. They can also be used for treating spleen and stomach weakness.

[0005] At present, there is no relevant research on the use of Jianwei Xiaoshi Tablets in the treatment of irritable bowel syndrome. SUMMARY

[0006] Therefore, the purpose of the present application is to provide the use of Jianwei Xiaoshi Tablets in the preparation of a medicine for treating irritable bowel syndrome. From the aspects of intestinal mucosal barrier and intestinal flora, the present application explores the action pathway and target of Jianwei Xiaoshi Tablets in the treatment of liver stagnation and spleen deficiency IBS-D rats, and analyzes the intestinal microecological diversity of rats after treatment, and explores the influence of Jianwei Xiaoshi Tablets on the intestinal flora changes of liver stagnation and spleen deficiency IBS-D rat models.

[0007] In order to achieve the above-mentioned purpose of the application, the technical scheme of the present application is as follows:

[0008] The present application provides the use of Jianwei Xiaoshi Tablets in the preparation of a medicine for treating irritable bowel syndrome.

[0009] That is, the present application provides Jianwei Xiaoshi Tablets for treating irritable bowel syndrome.

[0010] Preferably, the irritable bowel syndrome is liver stagnation and spleen deficiency diarrhea type irritable bowel syndrome.

[0011] Further preferably, the treatment of irritable bowel syndrome includes improving body weight, food intake, diarrhea and / or visceral sensitivity.

[0012] Further preferably, the treatment of irritable bowel syndrome comprises modulating the intestinal flora structure, inhibiting harmful bacteria and / or promoting the growth of beneficial bacteria.

[0013] Further preferably, the treatment of irritable bowel syndrome comprises promoting the expression of intestinal tight junction proteins Claudin-1, ZO-1 and / or MUC2.

[0014] Further preferably, the treatment of irritable bowel syndrome comprises improving the intestinal mucosal barrier damage.

[0015] Further preferably, the treatment of irritable bowel syndrome simultaneously comprises improving body weight, food intake, diarrhea condition and visceral sensitivity, modulating the intestinal flora structure, inhibiting harmful bacteria, promoting the growth of beneficial bacteria, promoting the expression of intestinal tight junction proteins Claudin-1, ZO-1 and MUC2 and improving the intestinal mucosal barrier damage.

[0016] Preferably, the stomachic and digestion-aiding tablet consists of Pericarpium Citri Reticulatae, Radix Pseudostellariae, Fried Wheat Germ, Fructus Crataegi and Rhizoma Dioscoreae.

[0017] In some specific embodiments, the stomachic and digestion-aiding tablet consists of, by weight parts, 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Wheat Germ, 100-120 parts of Fructus Crataegi and 100-190 parts of Rhizoma Dioscoreae.

[0018] Further preferably, the stomachic and digestion-aiding tablet consists of, by weight parts, 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Wheat Germ, 100-120 parts of Fructus Crataegi and 100-120 parts of Rhizoma Dioscoreae.

[0019] Further preferably, the stomachic and digestion-aiding tablet consists of, by weight parts, 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Wheat Germ, 100-120 parts of Fructus Crataegi and 160-190 parts of Rhizoma Dioscoreae.

[0020] Further preferably, the stomachic and digestion-aiding tablet consists of, by weight parts, 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Wheat Germ, 100-120 parts of Fructus Crataegi and 170-180 parts of Rhizoma Dioscoreae.

[0021] Further preferably, the stomachic and digestion-aiding tablet consists of, by weight parts, 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Wheat Germ, 100-120 parts of Fructus Crataegi and 170-180 parts of Rhizoma Dioscoreae.

[0022] The invention is: the prince ginseng is the monarch drug, and it is tonifying and nourishing; the dried tangerine or orange peel and the Chinese yam are the minister drugs, and they are tonifying and drying; the malt and the hawthorn are the auxiliary drugs, and they are digesting and tonifying the spleen; the combination of the above-mentioned traditional Chinese medicines can play the functions of tonifying qi, clearing and tonifying, soothing the liver and relieving depression, and invigorating the spleen and drying dampness, and the curative effect is remarkable for treating the irritable bowel syndrome of the diarrhea type of the liver depression and the spleen deficiency syndrome.

[0023] It should be noted that the "stomachic and digestant tablets" of the present invention are Chinese patent medicine, and those skilled in the art can understand that the stomachic and digestant tablets described in the present invention are not limited to tablets, and can be any dosage form of pharmaceutical composition with the same raw material components as the stomachic and digestant tablets of the present invention.

[0024] The present invention has the following beneficial effects:

[0025] The present invention provides the application of the stomachic and digestant tablets in the preparation of the medicine for treating the irritable bowel syndrome, expands the medical use of the stomachic and digestant tablets, and provides a new treatment idea for the irritable bowel syndrome, especially the irritable bowel syndrome of the diarrhea type of the liver depression and the spleen deficiency syndrome. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 is a schematic diagram of the general situation of rats in each group, note: A is the body weight change of rats in each group; B is the food intake change of rats in each group; compared with the NC group; #: P<0.05, ##: P<0.01, ###: P<0.001; compared with the MC group, *: P<0.05, **: P<0.01, ***: P<0.001.

[0027] Figure 2 is a schematic diagram of the fecal character change of rats in each group; note: A is the Bristol fecal character score; B is the water content of feces; compared with the NC group; #: P<0.05, ##: P<0.01, ###: P<0.001; compared with the MC group, *: P<0.05, **: P<0.01, ***: P<0.001.

[0028] Figure 3 is a behavioral observation diagram of rats in each group, note: A is the comparison of pain threshold; B is the comparison of sugar water preference; compared with the NC group; #: P<0.05, ##: P<0.01, ###: P<0.001; compared with the MC group, *: P<0.05, **: P<0.01, ***: P<0.001.

[0029] Figure 4 is the expression diagram of colon tissue HE staining, colon, Claudin-1, ZO-1 and MUC2 of rats in each group.

[0030] Figure 5 is a PCoA diagram of intestinal flora analysis of rats in each group, note: A is a PCoA diagram of three groups; B is a PCoA comparison diagram of the MC group and the XSP group.

[0031] Figure 6 is a cluster heat map analysis of the intestinal flora richness of each group of rats. Note: A is a cluster heat map analysis of species richness at the door level. Among them, the NC group from bottom to top is other species, Proteobacteria, Patescibacteria, Verrucomicrobia, Bacteroidetes, Firmicutes; the MC group and XSP from bottom to top are others, Proteobacteria, Verrucomicrobia, Bacteroidetes, Firmicutes; B is a cluster heat map analysis of species richness at the genus level. Among them, the NC group from bottom to top is others, Candidatus_Saccharimonas, Lachnoclostridium, norank_f_Lachnospiraceae, Akkermansia, norank_f_Muribaculaceae, Lachnospiraceae_NK4A136_group, unclassified_f_Lachnospiraceae, Bacteroides, Lactobacillus; the MC group and XSP from bottom to top are other species, Lachnoclostridium, norank_f_Lachnospiraceae, Akkermansia, norank_f_Muribaculaceae, Lachnospiraceae_NK4A136_group, unclassified_f_Lachnospiraceae, Bacteroides, Lactobacillus.

[0032] Figure 7 is an LDA Score (linear discriminant) analysis diagram of the intestinal flora of each group of rats. DETAILED DESCRIPTION

[0033] The following non-limiting examples can enable those of ordinary skill in the art to more fully understand the present application, but in no way limit the present application. The following is only an exemplary description of the scope of the present application, and those skilled in the art can make various changes and modifications to the present application according to the disclosed content, and it should also belong to the scope of the present application.

[0034] The present application will be further described below in the form of specific examples. The various chemical reagents used in the embodiments of the present application are obtained through conventional commercial channels unless otherwise specified. If not specified, the content described below is the mass content. If not specified, it is understood to be carried out at room temperature.

[0035] 1. Materials and methods

[0036] 1.1 Materials and reagents

[0037] Experimental animals: 30 SPF level 6-8 week old SD male rats, body weight about 200±20g, purchased from Zhejiang Vito Lihua Experimental Animal Technology Co., Ltd [production license number: SCXK(Zhe)2022-0001], animal feeding temperature (20±2) ℃, normal day and night change (12h / 12h). Animals were allowed to eat and drink freely during the experiment.

[0038] Stomach tablet from Jiangzhong Pharmaceutical Co., Ltd. is composed of Pericarpium Citri Reticulatae, Radix Pseudoginseng, Fried Malt, Fructus Crataegi and Rhizoma Dioscoreae. As an example, the stomach tablet is composed of 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudoginseng, 160-190 parts of Fried Malt, 100-120 parts of Fructus Crataegi and 100-120 parts of Rhizoma Dioscoreae by weight. As a specific example of the following examples, it is composed of 22.9 parts of Pericarpium Citri Reticulatae, 228.6 parts of Radix Pseudoginseng, 171.4 parts of Fried Malt, 114.3 parts of Fructus Crataegi and 171.4 parts of Rhizoma Dioscoreae by weight.

[0039] Spleen and liver Chinese medicine composition: Radix Codonopsis 15g, Poria cocos 20g, Radix Saposhnikoviae 10g, Fried Atractylodes 15g, Radix Bupleuri 10g, Rhizoma Cyperi 15g, Radix Aucklandiae 10g, Radix Curcumae 10g, Pericarpium Citri Reticulatae 15g, Radix Paeoniae Alba 10g, Fried Malt 10g, Fructus Aurantii 10g, Radix Glycyrrhizae 10g, purchased from Jiangxi University of Traditional Chinese Medicine Hospital of Traditional Chinese Medicine, the medicinal materials were washed and soaked in warm water for 30 min, decocted 3 times, concentrated to 1g of raw medicine / 1ml of medicinal liquid, filtered, sterilized and packaged, stored at 4℃ for standby. Senna leaf extract: Senna leaf decoction pieces (purchased from Jiangxi University of Traditional Chinese Medicine Hospital of Traditional Chinese Medicine) were soaked in 6 times the amount of boiling water for 30 min, repeated 3 times, then the mixed extract was filtered, concentrated at 50℃ under low pressure with a rotary evaporator, and the resulting liquid with a raw drug concentration of 0.45g / mL was transferred to a 4℃ refrigerator for use.

[0040] 1.2 Instruments and equipment

[0041] Multifunctional enzyme marker (PerkinElmer, USA), low-temperature high-speed centrifuge (ThermoFisher scientific, USA), small animal gas anesthesia machine (Shanghai Yurui Scientific Instrument Co., Ltd.), electronic balance (Beijing Sartorius Instrument System Co., Ltd.).

[0042] 1.3 Experimental methods

[0043] 1.3.1 Animal model establishment and drug administration

[0044] After 1 week of acclimation, the experimental animals were randomly divided into normal control (NC) group, model control (MC) group, Shugan Jianpi Chinese medicine (ZY) group and Xiangyansu tablet (XSP) group, 10 rats in each group. In this study, the rats were subjected to compound modeling: senna solution gavage + chronic restraint + tail clip stimulation, and the modeling time lasted for 14 days. Except for the NC group, the rest of the rats were given senna solution (0.45 g / mL) by gavage every day, with a gavage volume of 10 mL / kg / d, lasting for 14 days; 1 hour after gavage of senna solution, the rest of the rats were placed in the restraint device for 2 hours, once in the morning and once in the afternoon, lasting for 14 days; after the restraint, the tail clip was clamped at the 1 / 3 of the rat's tail, for 0.5 hours every day, lasting for 14 days. After modeling, the ZY group and the XSP group were given 0.84 g / kg of Shugan Jianpi Chinese medicine and Xiangyansu tablet raw material powder by gavage every day, and the NC group and the MC group were given the same amount of pure water, lasting for 14 days. Before modeling (day 0), after modeling (day 14), and after treatment (day 28), the body weight of rats in each group was measured, and the food intake of rats was recorded every day.

[0045] 1.3.2 Fecal consistency score

[0046] During the experiment, the rat feces were collected and observed, and their properties and appearance were evaluated according to the Bristol Stool Scale scoring standard to compare the fecal consistency scores of rats in each group. The fecal score of rats is as follows: 1 point: scattered hard lumps, like nuts; 2 points: sausage-like, but in chunks; 3 points: sausage-like, but with surface cracks; 4 points: like sausage or snake, smooth and soft; 5 points: soft mass, clear edge; 6 points: fluffy, unclear edge, paste-like stool; 7 points: water-like, no solid material.

[0047] 1.3.3 Fecal water content

[0048] The feces produced by each rat in an empty container for 2 hours were collected, weighed with aluminum foil shortly after collection, and then re-measured after drying overnight at 80°C. The percentage of water was obtained according to the ratio of dry weight / pre-baking weight.

[0049] Fecal water content = (wet feces weight - dry feces weight) / wet feces weight x 100%.

[0050] 1.3.4 Visceral sensitivity evaluation

[0051] Each group was subjected to colorectal distension according to the AWR scoring criteria, and the pain value was measured. The rats were given water and fasted within 12 hours before the experiment, and were given abdominal massage to promote defecation. After ether anesthesia, a small amount of paraffin oil was applied to the balloon in front of the 8Fr (diameter 3mm) catheter, the catheter was slowly inserted into the rat anus (about 6cm in length), and then the catheter was fixed on the rat tail with medical tape. The rats were placed in a transparent plastic box and kept still. After the rats were fully awake and adapted to the environment, the balloon was inflated, the first time was slowly inflated at a pressure of 10mmHg, and the second time was slowly inflated at a pressure of 20mmHg. Increase by 20mmHg / each time until the rat abdominal wall is observed to retract and lift from the bottom of the box. Each rat was measured 3 times, with an interval of 10min. The average pressure value of the three AWR scores was taken as the pain threshold of the rat. The lower the abdominal wall retraction reflex pain threshold, the higher the sensitivity of the rat to rectal balloon stimulation, and vice versa. The specific criteria for AWR score are as follows: 0 points, the rat has no behavioral response to colorectal distension; 1 point, the rat's head movement is reduced but the body is still stationary; 2 points, the rat's abdominal muscle contracts but the abdomen does not leave the table; 3 points, the rat's abdominal muscle contracts and the abdomen leaves the table; 4 points, the rat's pelvis is lifted and accompanied by a body arch.

[0052] 1.3.5 Sugar water preference test

[0053] The sugar water preference test was performed on the 14th day of administration. During the test, the rats were housed individually and each rat was given 2 bottles of 1% sucrose solution. After 24 hours, one of the bottles of sucrose solution was replaced with drinking water for another 24 hours, and then the rats were deprived of water for 23 hours. Thereafter, each rat was given a certain amount of 1 bottle of drinking water and 1 bottle of 1% sucrose solution, and the masses of the drinking water and 1% sucrose solution bottles were measured after 1 hour, respectively. The corresponding intake was calculated. Finally, the percentage of sugar water preference of the rats was calculated according to the formula.

[0054] Sugar water preference percentage = sugar water intake / (sugar water intake + drinking water intake) x 100%.

[0055] 1.3.6 HE staining

[0056] The colon fixed in 4% paraformaldehyde was dehydrated with ethanol from low concentration to high concentration, then placed in xylene for transparency, and then the transparent tissue was placed in paraffin. After the paraffin completely embedded the tissue, it was cooled. Then the embedded paraffin block was sectioned. The prepared section was placed in a 65°C oven and baked for 2h; then it was deparaffinated and washed with deionized water. First, it was stained with appropriate amount of hematoxylin for 5min, washed with water for 5s to ensure no residual dye on the section; then it was stained with appropriate amount of eosin for about 25s; finally, it was washed with color enhancer for 2 times, dried the excess liquid, and ensured the section was dry; it was mounted with neutral resin, photographed, and observed the morphological changes of the rat colon mucosa.

[0057] 1.3.7 Immunohistochemical method for detecting the expression of ZO-1, MUC2 and Claudin1 in colon tissue

[0058] After the above prepared section was deparaffinized, it was subjected to high temperature antigen retrieval, serum blocking, primary antibody incubation at 4°C overnight, and secondary antibody incubation at 37°C. After DAB color development, the section was placed in hematoxylin staining solution for restaining, gradient alcohol dehydration, xylene transparency, neutral resin mounting, and photographing.

[0059] Tight junction (TJ) is composed of multiple tight junction protein molecules, among which Claudin-1, ZO-1, Occludin, and MUC2 protein play a crucial role in maintaining its normal structure and function. The characteristic of intestinal barrier disruption is the reduced expression of ZO-1, MUC2, and Claudin1. Inhibiting the expression of Claudin, ZO-1, and other genes can effectively reverse the hyperalgesia of rats and the intestinal nervous system has a large number of neurons with a tight structure. The destruction of TJ may lead to repeated stimulation of the intestinal nervous system by food antigens, microorganisms, and their metabolites in the rat intestinal lumen.

[0060] 1.3.8 High-throughput sequencing of intestinal microorganism 16S rRNA gene

[0061] The 0.2 g rat fecal sample was taken for total microbial community DNA extraction and PCR amplification of the V3-V4 region of the 16S rRNA gene. The Miseq PE300 platform of Illumina was used for sequencing. The raw data was quality controlled and the sequences were optimized. The non-redundant sequences were clustered into OTUs according to 97% similarity in the clustering process, and the chimeras were removed to obtain the representative sequences of OTUs. All optimized sequences were mapped to the representative sequences of OTUs, and sequences with similarity above 97% to the representative sequences were selected to generate an OTU table. Then, based on the OTU table, species classification was performed to generate a phylogenetic tree, and the relative abundance at each level was divided; the diversity of each library was analyzed according to the OTU table data.

[0062] 1.3.9 Statistical analysis of data

[0063] The data were analyzed using GraphPad Prism 9 statistical software, and the results of each group were expressed as mean ± standard deviation, P<0.05 indicating significant difference. Intergroup comparison of intestinal flora was performed using one-way ANOVA, and pairwise comparison was performed using Student's t-test value test.

[0064] 2 Results

[0065] 2.1 General condition of rats The results are shown in Figure 1.

[0066] Before modeling, the rats were in normal general condition, with flexible reaction and good activity state, and the hair was soft and lustrous. After modeling, compared with the NC group, the rats in the modeling group had significantly reduced activity, mental fatigue, dryness and irritability, perianal moisture with attached loose stools, and preferred to huddle together, with dry and dull hair color. After the end of administration, the rats in the XSP and ZY groups had improved mental and activity conditions, with softer hair than before, and the feces gradually formed, with cleaner perianal region than the MC group.

[0067] There was no statistically significant difference in body weight and food intake between the rats in each group before modeling, and the body weight and food intake of the rats were significantly lower than those in the NC group after 2 weeks of modeling (P<0.01, P<0.001). After 2 weeks of treatment, compared with the MC group, the body weight and food intake of the rats in the XSP group were significantly increased (P<0.05, P<0.01), and the food intake of the rats in the ZY group was also significantly increased (P<0.05). The results showed that the modeling method in this experiment could cause spleen deficiency in rats, which in turn affected their food intake and body weight, and the XSP group could significantly improve the food intake and body weight of rats.

[0068] 2.2 Changes in fecal characteristics of rats The results are shown in Figure 2.

[0069] Compared with the NC group, the rats in the MC group had obvious paste and water-like stool, mostly type 6 or 7, and the Bristol stool form score and fecal water content were significantly increased (P<0.01, P<0.001); compared with the MC group, the Bristol score of the rats in the XSP group was significantly improved (P<0.05), and the fecal water content was also significantly reduced (P<0.05); the Bristol score and fecal water content of the rats in the ZY group were slightly improved, but there was no significant difference compared with the MC group.

[0070] 2.3 The results of the behavioral observation of rats are shown in Fig. 3.

[0071] The pain threshold of the rats in the MC group was significantly lower than that in the NC group (P<0.01), and after treatment with Weikang Xiaoshi Tablets, the pain threshold of the rats in the XSP group was up-regulated (P<0.05), and the pain threshold of the rats in the ZY group was slightly up-regulated, but there was no significant difference compared with the MC group. The results showed that the modeling method used in this experiment could reduce the pain threshold of rats and increase the visceral sensitivity, and Weikang Xiaoshi Tablets could significantly increase the pain threshold of rats and reduce the visceral sensitivity. The results of the sugar water preference test showed that compared with the NC group, the percentage of sugar water preference of the rats in the MC group was reduced (P<0.001); compared with the MC group, the percentage of sugar water preference of the rats in the XSP and ZY groups was increased (P<0.01, P<0.05). The results showed that the modeling method made the rats present a depressive state, thereby reducing the sugar water preference rate, and Weikang Xiaoshi Tablets and traditional Chinese medicine could significantly improve the depressive state of rats.

[0072] 2.4 The results of HE staining and immunohistochemistry of rat colon tissue are shown in Fig. 4.

[0073] The results of HE staining showed that the colon tissue structure of the rats in the NC group was complete, the morphology was normal, the glands were arranged in order, the crypts were closely aggregated, the goblet cells were uniformly distributed, and no damage was observed. The intestinal tissue integrity of the rats in the MC group was destroyed, the boundaries of each layer were not obvious, the intestinal mucosa was necrotic and shed, the intestinal wall was significantly thickened, accompanied by different degrees of edema and inflammatory cell infiltration, and the intestinal tissue integrity of the rats in the XSP group was restored to a certain extent, the intestinal wall thickening and immune infiltration were reduced. The results of immunofluorescence showed that ZO-1, Claudin-1 and MUC2 were mainly expressed in the epithelial layer of the intestinal mucosa. Compared with the NC group, the protein expression of ZO-1 and MUC2 in the MC group was significantly reduced, and the protein expression in the XSP group was slightly increased.

[0074] 2.5 The results of rat intestinal flora analysis are shown in Figs. 5-7.

[0075] 16S rRNA sequencing results are shown in Figures 5-7, PCoA analysis showed that there was a large difference between the NC group and the MC, XSP group samples, and there was also a certain difference between the MC and XSP group samples (P = 0.04). At the door level, Firmicutes had the highest relative abundance and was the dominant genus. Compared with the NC group, the relative abundance of Firmicutes in the MC and XSP groups decreased, while the relative abundance of Bacteroidetes and Verrucomicrobia increased significantly. At the genus level, there was a large difference between the NC group and the MC and XSP groups. The abundance of Lachnoclostridium, Candidatus_Saccharimonas and other genera in the NC group was significantly reduced in the MC and XSP groups. LEfSe multi-level species difference discriminant analysis of intestinal flora showed that XSP could restore the decrease in the relative abundance of c_Gammaproteobacteria, o_Betaproteobacteriales, f_Burkholderiaceae and g_Parasutterella caused by modeling.

[0076] In summary, during the experiment, the NC group rats were in good mental state, while the modeling group rats had sparse and messy hair, dark color, decreased appetite, loose and pale stool, severe perianal fecal contamination, listless spirit, and liked to huddle. After being given restraint and tail clamping stress stimulation, they became irritable and emotional.

[0077] Compared with the MC group rats, the XSP group rats had obvious improvement in mental state and activity, and the food intake and body weight also increased significantly. The diarrhea symptoms were significantly improved, the water-like stool or unformed stool condition was reduced, the Bristol score and fecal water content were significantly down-regulated, and the visceral sensitivity and sugar water preference were improved. The above experimental results showed that Jianwei Xiaoshi Tablets treatment could improve the liver stagnation and spleen deficiency syndrome of IBS-D rats.

[0078] Jianwei Xiaoshi Tablets treatment improved the expression of ZO-1 and MUC2 in intestinal epithelial tissue, indicating that it was involved in the protection of intestinal barrier. The results of this study confirmed that Jianwei Xiaoshi Tablets could effectively protect the intestinal mucosal barrier, improve the intestinal cavity environment, and thus reduce visceral hypersensitivity.

[0079] Jianwei Xiaoshi Tablets could increase the structural composition of intestinal flora in IBS-D rats with liver stagnation and spleen deficiency. Further analysis of the fecal intestinal flora of rats with richness difference found that Jianwei Xiaoshi Tablets could restore the decrease in the relative abundance of some colonies caused by modeling.

[0080] Therefore, Jianwei Xiaoshi Tablets can improve the general condition, body weight, food intake, diarrhea condition and visceral sensitivity of IBS-D rats with liver stagnation and spleen deficiency, regulate the intestinal flora structure of IBS-D rats with liver stagnation and spleen deficiency, inhibit harmful bacteria, promote the growth of beneficial bacteria, promote the expression of intestinal tight junction proteins Claudin-1, ZO-1 and MUC2, improve the damage of intestinal mucosal barrier, and improve IBS-D with liver stagnation and spleen deficiency.

Claims

1. Use of Weixie Xiaoshi Tablets in the preparation of a medicament for treating irritable bowel syndrome.

2. The use according to claim 1, wherein, The irritable bowel syndrome is diarrhea type irritable bowel syndrome of liver stagnation and spleen deficiency syndrome.

3. Use according to claim 1 or 2, wherein, The Weixie Xiaoshi Tablets are composed of Pericarpium Citri Reticulatae, Radix Pseudostellariae, Fried Hordeum Vulgare, Fructus Crataegi and Rhizoma Dioscoreae.

4. The use according to claim 1 or 2, wherein, The Weixie Xiaoshi Tablets are composed of 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Hordeum Vulgare, 100-120 parts of Fructus Crataegi and 100-190 parts of Rhizoma Dioscoreae.

5. The use according to claim 1 or 2, wherein, The Weixie Xiaoshi Tablets are composed of 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Hordeum Vulgare, 100-120 parts of Fructus Crataegi and 100-120 parts of Rhizoma Dioscoreae.

6. The use according to claim 1 or 2, wherein, The Weixie Xiaoshi Tablets are composed of 15-25 parts of Pericarpium Citri Reticulatae, 210-240 parts of Radix Pseudostellariae, 160-190 parts of Fried Hordeum Vulgare, 100-120 parts of Fructus Crataegi and 160-190 parts of Rhizoma Dioscoreae.

7. The use according to claim 1 or 2, wherein, The Weixie Xiaoshi Tablets are composed of 22.9 parts of Pericarpium Citri Reticulatae, 228.6 parts of Radix Pseudostellariae, 171.4 parts of Fried Hordeum Vulgare, 114.3 parts of Fructus Crataegi and 171.4 parts of Rhizoma Dioscoreae.

8. The use according to any one of claims 1 to 7, wherein, The treatment of irritable bowel syndrome includes improving body weight, food intake, diarrhea condition and / or visceral sensitivity.

9. The use according to any one of claims 1 to 7, wherein, The treatment of irritable bowel syndrome includes regulating intestinal flora structure, inhibiting harmful bacteria and / or promoting the growth of beneficial bacteria.

10. The use according to any one of claims 1 to 7, wherein, The treatment of irritable bowel syndrome includes promoting the expression of intestinal tight junction proteins Claudin-1, ZO-1 and MUC2.

11. The use according to any one of claims 1 to 7, wherein, The treatment of irritable bowel syndrome includes improving intestinal mucosal barrier damage.

12. The use according to any one of claims 1 to 11, wherein, The treatment of irritable bowel syndrome simultaneously includes improving body weight, food intake, diarrhea condition and visceral sensitivity, regulating intestinal flora structure, inhibiting harmful bacteria, promoting the growth of beneficial bacteria, promoting the expression of intestinal tight junction proteins Claudin-1, ZO-1 and MUC2 and improving intestinal mucosal barrier damage.

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