Study On The Microecological Mechanism Of Weifuchun In The Treatment Of Gastric Intestinal Metaplasia

Objective:Weifuchun is an effective Chinese patent medicine for the treatment of gastric intestinal metaplasia(GIM),but its mechanism of action is still unclear.Based on the tongue coating and gastric mucosa flora,this study explored the microecological mechanism of Weifuchun in the treatment of GIM,and combined with network pharmacology to explore its pharmacological mechanism.Methods:39 GIM patients were treated with Weifuchun for 4 weeks,while 26 patients with chronic superficial gastritis(CSG)served as controls.16S rDNA gene high-throughput sequencing was used to analyze tongue coating bacteria;the levels of serum pepsinogen I and II were detected using the latex immunoturbidimetric assay(LIA);an enzyme-linked immunosorbent assay(ELISA)was employed to detect the levels of serum IL-17,trefoil factor I,and TGF-α;a microplate-based quantification kit was utilized to detect serum total bile acid(TBA).37 GIM patients were recruited with Weifuchun for 4 weeks,while 26 patients with CSG served as controls.16S rDNA gene high-throughput sequencing was used to analyze gastric mucosa bacteria.Network pharmacology was used to analyze the active chemical components and targets of Weifuchun.Combined with the target of GIM,the interaction network of "Weifuchun-GIM" was constructed and topological analysis was carried out to screen core compounds and targets,and then KEGG pathway enrichment analysis was carried out.Results:(1)Weifuchun can reduce the diversity of tongue coating flora.Following the treatment,the relative abundances of tongue coating flora Granulicatella,Gemella,lachnoanerobaculum and Neisseria increased significantly,while the relative abundances of Alloprevotella,Prevotella,[Eeucterium]nodatum group and Ruminococceae UCG-014 increased significantly.Linear discriminant analysis(LDA)results showed that Alloprevotella and three rare tongue coating genera(Lautropia,Treponema 2,and Aliihoeflea)might be potential markers or target flora for treatment of GIM.KEGG function prediction analysis showed that Weifuchun might regulate bile secretion and folate biosynthesis in GIM patients by inhibiting flagellar assembly,peptidoglycan biosynthesis,and bacterial chemotaxis.The level of serum trefoil factor I decreased significantly in response to Weifuchun treatment,which is consistent with the decrease of folate biosynthesis predicted by KEGG.(2)Weifuchun can increase the richness and diversity of gastric mucosa flora in patients with GIM.Following the treatment,the relative abundances of gastric mucosa bacteria Helicobacter;Prevotella7,Lachnoanerobaculum,Neisseria,Acinetobacte,Streptococcu,Prevotella,Veillonella,Porphyromonas,Haemophilus,Fusobacterium,Alloprevotella and Granulicatella increased significantly,while the relative abundances of Ralstonia,Mesorhizobium,Pseudonocardia,Bradyrhizobium,Hydrobacter,Pseudolabrys,Variovorax,Reyranella,Aliidiomarina and Acidovorax decreased significantly(P<0.05).LDA results showed that Acinetobacter,Fusobacterium and thriteen rare tongue coating genera(Leptotrichia,Candidatus Saccharimonas,Actinomyces,Corynebacterium,Corynebacterium,Rothia,Aliihoeflea,Atopobium,Sphingobium,Sediminibacterium,Phycicoccus,Selenomonas 3 and Pseudarthrobacter)might be potential markers or target flora for treatment of GIM.KEGG function prediction analysis showed that Weifuchun might be related to the improvement of ABC transporters and transporters pathway,and the inhibition of DNA repair and recombination proteins,ribosome and DNA replication Proteins,which are involved in DNA damage,repair and translation pathway,as well as the inhibition of enzymes and amino sugar and nucleotide sugar metabolism.(3)Network pharmacology analysis showed that 7-hydroxycoumarin,amethystoicacid,epoxyclimonin,quercetin,naringenin and protopanaxatriol were the key compounds of Weifuchun treatment of GIM.These compounds through 16 key targets(MAP2k1,JAK2,VEGFA,PIK3R1,et al).KEGG enrichment analysis indicated that the target of Weifuchun-GIM involved 156 pathways.Weifuchun GIM target pathway has 8 intersections with KEGG function prediction analysis of tongue coating flora,and 1 intersection with KEGG function prediction analysis of gastric mucosa flora pathway-ABC transporter.Conclusion:This study found that the changes of tongue coating and gastric mucosa flora are related to the effective treatment of GIM by Weifuchun,suggesting that Weifuchun may achieve the therapeutic effect by regulating the balance of gastrointestinal flora.At the same time,network pharmacology analysis suggests that Weifuchun can treat GIM through multi-component,multi-target and multi-channel treatment.It provides a new idea for the follow-up animal experiments to reveal the molecular biological mechanism of Weifuchun in the treatment of GIM.