基于生物信息学技术和实验验证探讨肝胃百合汤治疗胃溃疡作用机制

Discussion on the mechanism of Ganwei Baihe Decoction in treating gastric ulcer based on bioinformatics and experimental validation

目的基于生物信息学研究肝胃百合汤治疗胃溃疡的潜在作用机制,并通过动物实验进行验证。方法利用TCMSP、DisGeNET、GeneCards数据库检索肝胃百合汤活性成分、作用靶点及胃溃疡作用靶点,取交集后通过STRING数据库获取交集靶点蛋白相互作用数据,并采用Cytoscape 3.10.0软件构建PPI网络,获取关键靶点和关键成分,利用DAVID在线分析数据库对关键靶点进行GO功能和KEGG通路富集分析。通过动物实验进行验证。将36只SD大鼠按随机数字表分为空白组、模型组、奥美拉唑组、肝胃百合汤组,每组9只。各组大鼠灌胃相应药物7 d后,禁食不禁水24 h,复灌胃1次,给药1 h后灌胃吲哚美辛80 mg/kg制备胃溃疡大鼠模型,给药3 h后麻醉取材,采用Western blot法测定大鼠胃组织Caspase-3蛋白表达。结果获得肝胃百合汤活性成分234个,靶点290个,治疗胃溃疡作用靶点6496个,筛选出肝胃百合汤治疗胃溃疡潜在靶点213个。GO功能富集条目437条,KEGG通路富集条目153条。与模型组比较,肝胃百合汤组、奥美拉唑组Caspase-3蛋白表达降低(P<0.05)。结论肝胃百合汤治疗胃溃疡作用机制可能是通过槲皮素、β-谷甾醇等关键成分,作用于AKT1、CASP3等关键靶点,调控Apoptosis通路、PI3K-Akt信号通路、MAPK信号通路等发挥抑制凋亡作用。

Objective To study the potential mechanism of Ganwei Baihe Decoction in the treatment of gastric ulcer(GU)based on bioinformatics and validate it through animal experiments.Methods TCMSP,DisGeNET,and GeneCards databases were used to retrieved active components and action targets of Ganwei Baihe Decoction.After obtaining the intersection,protein interaction data of the intersection genes were obtained through the STRING database.A PPI network was constructed by Cytoscape 3.10.0 software and the key genes and key components were obtained.DAVID online analysis database was used for GO functional enrichment and KEGG pathway enrichment analysis of key targets.Animal experiments were used for verification.Totally 36 SD rats were divided into blank group,model group,Omeprazole group and Ganwei Baihe Decoction group according to the random number table method,with 9 rats in each group.After 7 days of gavage of the corresponding drugs to each group of rats,they fasted and but with water for 24 hours,and then re-gavaged once.After 1 hour of administration,a gastric ulcer rat model was prepared by gavage of 80 mg/kg of indomethacin.After 3 hours of administration,anesthesia was used to extract the sample.The expression level of Caspase-3 protein in the gastric tissue of rats was to be determined by Western blot method.Results There were 234 effective active components with 290 targets in Ganwei Baihe Decoction,and 6496 therapeutic targets for GU.213 potential targets for GU were screened out.There were 437 GO function and 153 KEGG pathway enriched entries.Compared with the model group,the protein expression of Caspase-3 in the Ganwei Baihe Decoction group and Omeprazole group decreased(P<0.05).Conclusion The mechanism of Ganwei Baihe Decoction in treating GU may be through key components such as quercetin andβ-sitosterol acting on key targets such as AKT1 and CASP3,regulating the Apoptosis pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,etc.to exert inhibitory effects on apoptosis.