基于网络药理学和实验验证探讨胃复方对胃癌的效应及作用机制

Experimental verification of the effect and mechanism of Weifufang on gastric cancer based on network pharmacology

目的 基于网络药理学探究胃复方治疗胃癌的作用机制。方法 应用中药系统药理学数据库及分析平台、DrugBank、GeneCards等数据库检索胃复方所含12味中药的有效化学成分及治疗胃癌的潜在靶点,将潜在靶点导入DAVID数据库进行基因本体论(GO)及京都基因与基因组百科全书(KEGG)分析,筛选出潜在的生物过程和涉及的信号通路。采用CCK-8及Western blotting对KEGG分析富集程度最大的信号通路进行验证。结果 胃复方治疗胃癌可能涉及以槲皮素、木樨草素为代表的88种有效活性成分,可能作用于以TP53、STAT3、Akt为代表的194个靶点。GO分析涉及148个生物过程,KEGG分析筛选出PI3K/Akt、TNF、HIF-1等信号通路。体外实验证实胃复方提取物可抑制人胃癌细胞HGC-27增殖,降低其p-PI3K、PI3K、p-Akt、Akt蛋白表达水平;而对于人正常胃黏膜上皮细胞GES-1,胃复方提取物可促进其增殖,上调其p-PI3K、PI3K、p-Akt、Akt蛋白表达。结论 本研究初步预测了胃复方治疗胃癌“中药-多成分-多靶点-多通路”的调控网络,并验证了胃复方提取物可能通过抑制PI3K/Akt信号通路相关蛋白的表达发挥抑制胃癌细胞增殖的作用,证明了网络药理学技术挖掘胃复方具有一定的可信度,但同时兼有不全面性,为进一步探索胃复方的作用机制奠定了基础。

Objective To explore the mechanism of Weifufang(an experienced effective prescription for the treatment of gastric cancer developed by Hunan Cancer Hospital)in treating gastric cancer on the base of network pharmacology.Meth-ods The effective active ingredients and potential targets for treating gastric cancer of 12 traditional Chinese medicines con-tained in Weifufang were searched by using the Chinese Medicine System Pharmacology Database and Analysis Platform,DrugBank,GeneCards and other databases.The potential targets were imported into DAVID database for gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis,and the potential biological processes and involved signal pathways were screened out.CCK-8 and Western blotting were used to verify the signal pathway with the highest enrichment degree in KEGG analysis.Results Weifufang therapy for gastric cancer may involve 88 active ingredi-ents represented by quercetin and luteolin,and may act on 194 targets represented by TP53,STAT3 and Akt.GO analysis involved 148 biological processes,and KEGG analysis screened PI3K/Akt,TNF,HIF-1 and other signal pathways.In vitro experiments proved that the Weifufang extract could inhibit the proliferation of human gastric cancer HGC-27 cells,and decrease the protein expression of p-PI3K,PI3K,p-Akt and Akt.However,Weifufang extract could promote the prolifera-tion of human normal gastric epithelial cells GES-1 and up-regulate the protein expression of p-PI3K,PI3K,p-Akt and Akt.Conclusion This study preliminarily predicted the regulatory network of"traditional Chinese medicine-multicompo-nents-multitargets-multipathways"in the treatment of gastric cancer,and verified that the Weifufang extract could inhibit the proliferation of gastric cancer cells by inhibiting the expression of PI3K/Akt signal pathway related proteins.It proved that the network pharmacology technology was credible in mining Weifufang,but it was incomplete at the same time,provid-ing a basis for further exploring the mechanism of Weifufang.