Research On The Therapeutic Effect And Mechanism Of Qingwei Zhitong Pills Induced By Acetic Acid Gastric Ulcer In Rats

Gastric ulcer(GU),a common digestive system disease,is a type of peptic ulcer occurring on the lining mucosa of the stomach or at a deeper level.GU has the characteristics of high incidence and long course of disease.Stomach burning pain is the typical clinical manifestation of GU,often accompanied by loss of appetite,postprandial abdominal distension and other gastrointestinal symptoms.In severe cases,it can be accompanied by bleeding,perforation,pyloric obstruction,gastric cancer and other serious complications.The current clinical drugs for GU have shortcomings such as drug resistance and side effects,So it is very important to develop new therapeutic drugs.Traditional Chinese medicine and its compound have the advantages of multiple targets and few side effects.Qingwei Zhitong Pellet is a traditional Chinese medicine compound for the treatment of GU,which is composed of coptidis,Paeoniae alba,sanguinae ulmus,Baihe and Radix japonicus.Although Qingwei Zhitong pills have good therapeutic effect in clinic,the mechanism of improving GU is still unclear.Therefore,this study intends to verify and evaluate the pharmacodynamic effects of Qingweitong pills on GU at the animal level through the animal model of acetic acid GU rats.The key target of Qingwei Zhitong pills in the treatment of acetic acid type GU rats was predicted by network pharmacology.Based on microbiome and metabolomics analysis,the protective mechanism of Qingweitong pills against GU was revealed from a new perspective of microbiota and metabolism,and drug targets predicted by network pharmacology and metabolic pathways screened by metabolomics were combined for analysis,which could provide more scientific data for clinical application of Qingweitong pills and promote the secondary development process of traditional Chinese patent medicine.Objective:To study the therapeutic effects of Gastroparesis Micro Pills on acetic acid gastric ulcer in rats,to analyze and identify the small molecule chemical components in Gastroparesis Micro Pills,and to predict the key components and targets of Gastroparesis Micro Pills and their involved signaling pathways based on network pharmacology,and to provide scientific data for the elucidation of its chemical composition and mechanism of action.To provide scientific data to elucidate the chemical composition and mechanism of action.Microbiomics and metabolomics techniques were used to analyze the specific metabolites and explore their possible mechanisms of action.Methods:Forty-eight male WISTAR rats were divided into blank group,gastric ulcer model group,clear stomach pain relief micro-pill Yang high,medium and low(250,125 and 62.5 mg/ml)dose groups and omeprazole positive treatment control group(0.26 mg/ml),8 rats in each group.The WISTAR murine GU model was prepared by acetic acid injection,and the normal group was left untreated.The other groups were anesthetized by intraperitoneal injection with 10%chloral hydrate(3 ml/kg)on the day before and the day of surgery with 24 hours of fasting and water abstinence.The upper abdominal part of the rats was prepared,disinfected,and surgical cavity towels were laid,and the stomach was exposed by a longitudinal incision from 1.5-2.0 cm above the glabella,and the stomach was injected with 0.3 ml of 30%acetic acid solution under the plasma membrane layer in the pyloric region of the anterior wall of the stomach(gastric-body junction)until there was no obvious bleeding,and then the anterior wall of the stomach was rinsed with saline twice,and then the stomach was delivered into the abdomen,and the peritoneum,muscle layer,and skin were sutured layer by layer,disinfected with iodine volt 3 times,and coated with Malone musk hemorrhoid cream to reduce inflammation.The gastric ulcer model group and the blank group were administered with saline gavage at a volume of 10 ml/kg,and the gastric pain relief micro-pill and omeprazole positive control group were administered continuously for 7 d.The chemical composition of gastric pain relief micro-pill and its targets were screened through the database,and the related targets of gastric ulcer were screened at the same time to obtain the intersection targets,so as to construct a "chemical composition-intersection target-gastric ulcer"interplay.-Gastric ulcer" interactions network,and screened the key chemical components that play an important role in the treatment of gastric ulcer in Gastroparesis Micro Pills;constructed protein interactions network,screened the key targets;used GO and KEGG enrichment analysis,and combined with related literature,screened the important signaling pathways.Finally,microbiomics 16S rDNA and metabolomics were used to search for its possible differential flora and metabolites,and also to search for relevant metabolic pathways that may affect the occurrence of acetic acid type GU,and to preliminarily predict the possible pathogenesis of GU.Results:1.Based on acetic acid type GU rat model,the pharmacodynamics of Qingwei Zhitong pills in the treatment of GU were evaluated,and the results showed that the weight of rats after Qingwei Zhitong pills was increased;Ulcer area decreased;The ulcer inhibition rates of low,medium and high dose groups were 22.72%,37.12%and 29.54%,respectively.HE results showed that the pathology of rats treated with Qingwei Zhitong pellets was significantly improved in each dose group,especially in the medium dose group.2.The possible targets of Qingwei Zhitong pellets in the treatment of acetic acid GU were analyzed by network pharmacology,including TNF,STAT3,HRAS,ESR1 and AKT1.Possible pathways of action include PI3K-Akt,MAPK and AGE/RAGE.3.Through the study of microbiome,it was found that the abundance of Cyanobacteria,Proteobacteria and Bacteroidota was higher in GU rats through a variety of statistical analysis.The abundance of GU rats in Firmicutes,Acidobacteria and Desulfobacterota is low;The abundance of Firmicutes,Bacteroidota and other bacteria increased in GU rats after being treated with Qingwei Zhitong pills.The abundance of Proteobacteria,Actinobacteriota and Acidobacteria decreased.Therefore,Cyanobacteria,Proteobacteria and Bacteroidota may be the potential pathogenic bacteria groups of GU,while Firmicutes may be beneficial to GU treatment and may be a potential therapeutic target.4.Metabolomics analysis showed that the serum metabolites level of acetic acid GU model rats(group M)was significantly different from that of blank group(group K).A total of 37 different metabolites were screened.A total of 33 different metabolites were screened in the treatment group(group H)compared with group M.Subsequent KEGG enrichment analysis showed that 12 metabolic pathways were screened out in group K compared with group M,and significant changes occurred in Arachidonic acid metabolism and Linoleic acid metabolism.A total of 10 metabolic pathways were screened out in group H compared with group M,among which Sphingolipid metabolism changed significantly.Finally,through the combined analysis of network pharmacology and metabolomics,three key metabolites were screened out in group H compared with group M:12(13)-EpOME,1-stearyl-Sn-gly cero-3-phosphocholine(1-acyl-Sn-Glycaro-3-phosphocholine),1,2-dihydroxynaphthalene(Naphehalene-1,2-diol);Two key targets:PTGS2 and CYP3A4;12 key metabolic pathways:Exogenous substance metabolism,tyrosine metabolism,tryptophan metabolism,anti-inflammatory metabolites possibly formed by EPA,prostaglandin formed by 2h-gamma-linoleic acid,prostaglandin formed by arachidonic acid,linoleate metabolism,leukotriene metabolism,glycerol phospholipid metabolism,C21-Biosynthesis and metabolism of steroid hormones,arachidonic acid metabolism,androgen and estrogen biosynthesis and metabolism.Conclusion:This study completed the pharmacodynamic evaluation of QingGastra pain relief micro-pills based on the acetic acid-induced gastric ulcer model;predicted the targets and their signaling pathways for the drug treatment of GU by network pharmacology technique;and then preliminarily elucidated that QingGastra pain relief micro-pills can play a role in the treatment of GU by restoring the structural dysbiosis of bacterial flora and metabolic disorders of the body based on microbiome and metabolome study techniques.This study provides further scientific data for the secondary development and clinical application of traditional Chinese medicines.