| Purpose:To investigate the mechanism of action of Jianpi Huayu Formula in the treatment of chronic atrophic gastritis(CAG)based on network pharmacology and molecular docking,to predict the drug targets of Jianpi Huayu Formula,to further conduct in vivo validation experiments in rats,to explore the therapeutic effects of Jianpi Huayu Formula on CAG rats,and to provide theoretical and experimental bases for the clinical promotion of Jianpi Huayu Formula.Material and method:Paper 1: Exploring the mechanism of action of Jian spleen and Huayu formula for the treatment of CAG based on network pharmacology and molecular dockingA "drug-component-target protein-disease" protein interaction network was constructed by predicting the action targets of Jianpi Huayu Formula and CAG through a network database,and the intersecting targets of disease and drug were subjected to gene ontology analysis(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The main biological functions and signalling pathways involved in CAG were identified,and the components with high network neutrality values were molecularly docked to the core targets.Paper 2: Effects of Jianpi Huayu Formula on p-AKT/AKT ratio,p-PTEN,PTEN and serum IL-17 and TNF-α in the gastric mucosa of CAG ratsSixty SD rats were randomly distributed into 4 groups(blank group,model group,Jianpi Huayu Formula group and Vitamin tablets group).Except for the blank group,all the groups were established as a rat CAG model according to the MNNG compound method[1] with a 12-week cycle.After the successful preparation of the model was determined by histopathological examination,the model group,the Jianpi Huayu Fang group and the Vitamin tablets group were started to be treated with double-distilled water,Jianpi Huayu Fang and Vitamin tablets,respectively,for a 12-week treatment period.After the completion of treatment,the general condition of the rats in each experimental group was observed;the gastrointestinal mucosa pathological changes of the rats were observed;the expression levels of AKT,p-AKT,PTEN and p-PTEN in the gastric mucosa tissues of the rats in each group were detected by Western blot;the serum IL-17 and TNF-α contents of the rats were detected by ELISA.SPSS26.0 was applied to analyze the data results,and Graph Pad prism 9.3.1 was applied to make graphs.Results:Paper 1: Exploring the mechanism of action of Jianpi Huayu Formula in the treatment of CAG based on network pharmacology and molecular docking1.A "drug-component-target protein-disease" protein interaction network was constructed and the core targets were identified as AKT1 and BCL2L1,and the core active ingredients were identified as quercetin,lignan and kaempferol.The GO analysis showed that the biological functions mainly involve oxidation reaction,and the molecular functions mainly involve ubiquitin-like protein ligase binding,etc.The KEGG analysis showed that the treatment of CAG by Jianpi Huayu Formula mainly involves PI3K/AKT signaling pathway,IL-17 signaling pathway and TNF signaling pathway,etc.3.Molecular docking verified that the top 3 compounds quercetin,lignan and kaempferol all bound better to the core targets AKT1 and BCL2L1.Paper 2: Effects of Jianpi Huayu Formula on p-PTEN,PTEN,p-AKT/AKT and serum IL-17 and TNF-α in gastric mucosa of CAG rats1.General condition of rats: The rats in the blank group were in normal general condition and in good mental condition.The rats in the model group showed slow weight gain,reduced food intake and water consumption compared with other groups,mental atrophy,reduced activity and lusterless fur.2.Gastric mucosa pathological changes: compared with the blank group,the model group rats showed atrophy of gastric mucosa(P<0.05);compared with the model group,the incidence of gastric mucosa atrophy in the Jianpi Huayu Fang group(P<0.05)and the Vitamin tablets group(P<0.05)was significantly reduced;in terms of gastric mucosa atrophy score,compared with the blank group,the model group rats had a higher atrophy score(P<0.05).),compared with the model group,the atrophy scores of the Jianpi Huayu Formula group(P<0.05)and the Vimycin group(P<0.05)were lower,but the difference between the two groups was not significant and the difference was not statistically significant.3.Gastric mucosal protein expression: there was no statistical difference in AKT and p-PTEN in the gastric mucosal tissues of rats in each group compared with each other;p-AKT/AKT expression: when it was compared to the blank group,the level of p-AKT/AKT(P<0.01)expression increased in the model group,and when it was divided into two groups,the level of p-AKT/AKT(P<0.05)expression decreased in the Jianpi Huayu Fang group and the differences were The expression PTEN(P<0.001)expression increased in the model group and PTEN(P<0.05)expression increased in the Vitamin tablets group,the discrepancies were statistically significant;compared with the Vitamin tablets group,the difference in expression in the Jianpi Huayu Fang group(p>0.05)was small and not statistically significant.4.Serum IL-17 and TNF-α levels: compared with the blank group,the levels of IL-17(P<0.001)and TNF-α(P<0.001)were increased in the model group,compared with the model group,the levels of IL-17(P<0.01)and TNF-α(P<0.05)were decreased in the Jianpi Huayu Fang group,and the levels of IL-17(P<0.01)and TNF-α(p <0.05)levels were all reduced,and the discrepancies were statistically significant;compared with the Vitamin tablets group,IL-17(P > 0.05)and TNF-α(P > 0.05)levels were slightly lower in the cancerblocking gastric tide group,but the differences were not statistically significant.Conclusion:1.The Jianpi Huayu Formula can inhibit the inactivation of PTEN in the gastric mucosa of CAG model rats and down-regulate the phosphorylation level of AKT,thus inhibiting the over-activation of PTEN/AKT pathway and playing an anti-apoptotic effect,which in turn reduces the atrophy of gastric mucosa.2.The Jianpi Huayu Formula can reduce the secretion of inflammatory factors IL-17 and TNF-α in the serum of CAG model rats,thus exerting an anti-inflammatory effect. |
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