Effect Of Yiqi Huoxue Intervention On Chronic Atrophic Gastritis And Gastric Precancerous Lesions

Objective:To investigate the effect of Yiqi Huoxue intervention on chronic atrophic gastritis(CAG)and gastric precancerous lesions(GPL)by collecting clinical cases.Based on the clinical collection of Chinese herbs prescriptions,we explored the drug rules of Yiqi Huoxue in treating CAG and GPL through R language data mining,to mine the core drug combination and optimize prescription compositions.Based on the core drug combination,the network pharmacology and molecular docking technology were used to explore the potential mechanism of Yiqi Huoxue against CAG and GPL,including the core active components,key target proteins,and main pathways.The target proteins were verified in gene chips and experimental study.Methods:1.Clinical study on the effect of Yiqi Huoxue intervention on CAG and GPLCollecting patients with CAG and GPL treated with Yiqi Huoxue decoction in Gastroenterology of Jiangsu Province Hospital of Chinese Medicine from 2016 to 2021.According to the endoscopic pathological changes of CAG and GPL before and after treatment,the effect of Yiqi Huoxue intervention on the long-term efficacy of CAG and GPL was evaluated from the aspects of overall pathohiological histology,pathological histological types,pathological histologic score,gastroscopic time,anatomical position and OLGA/OLGIM grading and staging,and the risk factors of CAG and GPL were explored.2.The medication rules of Yiqi Huoxue against CAG and GPL based on R language data miningBased on the case datasets collected from clinical research,a database of Chinese herbs prescriptions for the treatment of CAG and GPL with Yiqi Huoxue was established,and the prescriptions of Chinese herbs were extracted and processed in a standardized manner.Based on the functions of R language and its related packages,the frequency statistics of Chinese herbs,the statistics of properties and flavors,the statistics of main treatment efficacy,the analysis of association rules and cluster analysis were performed on the obtained Chinese herbs prescriptions.3.Exploring the Molecular Mechanism of Yiqi Huoxue against CAG and GPL by Network Pharmacology and Experimental StudyBaizhu-Dangshen-shijianchuan(BDS)active ingredients and the corresponding target proteins were acquired from the TCMSP database.Disease target proteins were collected from GeneCards,OMIM,DisGeNET,TTD and Drugbank databases.Venn diagrams were drawn and intersecting target proteins were obtained.The intersecting proteins were submitted to STRING v11.5 to construct the protein-protein interaction(PPI)network.Cytoscape 3.7.2 was used to visualize the results of the PPI network and construct the network diagram of drug-potential active ingredient-potential target protein-KEGG pathways.GO enrichment analysis and KEGG pathway analysis were conducted with R 4.0.3.Autodock 4.2.6 was used for molecular docking,and Pymol software was used to visualize the molecular docking results.Relevant tissue microarray data were obtained from the GEO database to verify key target proteins at the mRNA level.The gastric mucosal intestinal metaplasia model was constructed to verify the main active components and core target proteins.Results:1.A total of 300 patients were included in the efficacy evaluation.(1)Overall histopathology:The improvement rate,stability rate and progression rate of overall histopathology were 57.67%,34.67%and 7.67%,respectively.(2)Different histopathological types:In terms of improvement rate,the overall improvement rates of gastric atrophy(GA),intestinal metaplasia(IM)and low-grade intraepithelial neoplasia(LGIN)were 51.71%,48.10%and 76.09%,respectively,and the difference among the three types was statistically significant(P=0.002).There were statistically significant differences between LGIN improvement rate and GA improvement rate(P=0.002).The difference between the LGIN improvement rate and IM the improvement rate was also statistically significant(P=0.0004).In terms of stabilization rates,the overall stabilization rates of GA,IM,and LGIN were 39.38%,42.90%,and 23.91%,respectively,and the difference was statistically significant(P=0.049).There were statistically significant differences between LGIN stabilization rate and GA stabilization rate(P=0.044).The difference between the LGIN stabilization rate and IM the stabilization rate was also statistically significant(P=0.015).In terms of progression rate,the overall progression rates of GA,IM and LGIN were 8.90%,9.00%and 0.00%,respectively,and there was no significant difference(P>0.05).(3)Histopathological score:The pathological scores of GA,IM and LGIN were significantly decreased after treatment(P<0.05).(4)Different endoscopy time:≤12 months,the overall histopathological improvement,stability and progression rates were 61.59%,35.10%and 3.31%,respectively.12～36 months,the overall histopathological improvement,stability,and progression rates were 51.88%,36.09%,and 12.03%,respectively.36～60 months,the overall histopathological improvement,stability,and progression rates were respectively were 49.18%,47.54%and 3.28%,respectively.The improvement,stabilization and progression of different gastroscopy time were compared and analyzed,and there was no significant difference among the three time periods in terms of improvement rate and stabilization rate(P>0.05).In terms of progression rate,there was statistically significant difference between 12～36 months and ≤12 months(P=0.003).Meanwhile,statistically significant differences existed between 12～36 months and 36～60 months(P=0.043).(5)Different anatomical locations:There was no significant difference in the histopathological changes of antrum and corpus(P>0.05).(6)OLGA/OLGIM:The overall improvement rate,stability rate and progression rate of OLGA were 56.92%,26.15%and 16.92%,respectively,and the improvement rate of OLGA III-IV was significantly higher than that of OLGA stage I-II(P=0.018).The overall improvement rate,stability rate and progression rate of OLGIM were 50.00%,31.25%and 18.75%,respectively.There was no statistically significant difference between the two groups of patients in OLGIMⅠ-Ⅱ and OLGIM Ⅲ-Ⅳ(P>0.05).The two grading and staging methods were compared and analyzed,and there was no significant difference in OLGA/OLGIM in terms of improvement,stability and progression(P>0.05).(7)Risk factors:Univariate analysis showed that there were significant differences between the two groups in Helicobacter pylori infection,bile reflux and gastric mucosal erosion(P<0.05).Multivariate analysis showed that Helicobacter pylori infection,bile reflux and erosion of gastric mucosa were independent risk factors for CAG and GPL(P<0.05).(8)Stratified analysis:Stratified by Hp infection,the difference among the groups was statistically significant(P<0.05).2.A total of 468 Chinese herbs prescriptions for Yiqi Huoxue were obtained,including 142 kinds of CMH,and the total frequency was 8,005,of which Baizhu(465 times),Chenpi(460 times),Fuling(457 times),Gancao(412 times)and Yiyiren(405 times)were the top 5 most frequently used Chinese herbs.The properties of the medicine are mainly warm,the tastes of the medicine are mainly sweet,the main meridian of the medicine is the spleen meridian of foot taiyin,and the main therapeutic effects are the deficiency-nourishing herbs,the herbs for promoting blood circulation and removing blood stasis.Whole prescription association rule analysis showed that the combination of Baizhu,Dangshen,Chenpi,Fuling,Gancao,Ezhu,Shijianchuan,Huangqi,Yujin,Yiyiren,and Zhiqiao was the core drug combination.Whole prescription cluster analysis confirmed that the above 11 Chinese herbs could change the clustering relationship.The association rule analysis of Yiqi Huoxue Chinese herbs showed that the combination of Baizhu,Dangshen,Shijianchuan,and Huangqi was the core drug combination.The cluster analysis of Yiqi Huoxue Chinese herbs showed that Baizhu,Dangshen,and Shijianchuan could change the clustering relationship.3.A total of 30 active ingredients,353 Chinese herbs target proteins,3750 disease target proteins,and 228 intersection targets were obtained.Biological processes of Go enrichment analysis mainly involved regulation of protein serine/threonine kinase activity,peptidyl-serine phosphorylation,peptidyl-serine modification,epithelial cell proliferation,and positive regulation of anion transport.Molecular function of Go enrichment analysis mainly involved protein serine/threonine kinase activity,DNA-binding transcription factor binding,ubiquitin-like protein ligase binding,phosphatase binding,and ubiquitin protein ligase binding.Cellular component of Go enrichment analysis mainly involved transferase complex,transferring phosphorus-containing groups,transcription regulator complex,serine/threonine protein kinase complex,protein kinase complex,and membrane raft.KEGG pathway analysis mainly involved PI3K/Akt signaling pathway,virus infection,cancer,cell senescence and endocrine resistance.PI3K/Akt signaling pathway was the pathway with the largest number of gene enrichment.Finally,5 core active components and 26 key target proteins were obtained,of which quercetin and luteolin were the active components with the highest degree value,MAPK1,MAPK3,CCND1,AKT1 and TP53 were the key target proteins with the highest Degree values.Molecular docking showed that quercetin and luteolin had good docking activity with the corresponding key target proteins.The results of microarray analysis showed that twenty-one key target proteins were verified in various pathological tissues of different microarray expression profiling datasets.Experimental studies showed that the protein expression level of CASP3 in chenodeoxycholic acid(CDCA)-induced gastric mucosal intestinal metaplasia model was significantly decreased after quercetin intervention(P<0.05),and the protein expression levels of the intestinal metaplasia marker CDX2 and its downstream markers(KLF4,VIL1)were also significantly decreased(P<0.05).Conclusions:1.Yiqi Huoxue intervention improved the pathology of CAG and GPL,keeping lesions steady,and reducing disease progression.The clinical efficacy of Yiqi Huoxue intervention may not decrease significantly over time.Yiqi Huoxue intervention could produce better therapeutic effect on high-risk groups of gastric cancer(OLGA Ⅲ-Ⅳ).H.pylori infection,gastric mucosal erosion and bile reflux were independent risk factors for CAG and GPL.2.The combination of Baizhu,Dangshen,Chenpi,Fuling,Gancao,Ezhu,Shijianchuan,Huangqi,Yujin,Yiyiren,and Zhiqiao was the core drug combination of Yiqi Huoxue therapy in treating CAG and GPL and the combination of Baizhu,Dangshen and Shijianchuan was the core drug combination of Yiqi Huoxue Chinese herbs.3.BDS may act on multiple disease target proteins,such as CASP3,CCND1 and RB1,through various active components,such as quercetin and luteolin,and regulate PI3K/Akt and other signaling pathways to improve the pathological state of CAG and GPL,and prevent inflammatory cancer transformation.Quercetin may inhibit CDCA-induced gastric mucosa intestinal metaplasia by regulating CASP3.