葛花作用于Casp3调控细胞自噬干预肝癌的机制研究  

Mechanism of Pueraria lobata in regulating liver cancer via Casp3mediated cellular autophagy

作　　者：李军 成果 楼迪栋 张楠楠 LI Jun;CHENG Guo;LOU Didong;ZHANG Nannan(School of Basic Medical Sciences,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;The First Clinical Medical College,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]贵州中医药大学基础医学院,贵州贵阳550025 [2]贵州中医药大学第一临床医学院,贵州贵阳550025

出　　处：《海南医科大学学报》2025年第3期200-210,共11页Journal of Hainan Medical University

基　　金：贵州省基础研究计划自然科学项目[黔科合基础ZK(2024)一般382];2024年贵州省中管局项目(QZYY-2024-001);2023年贵州中医药大学研究生教育创新计划项目(YCXKYB2023001)。

摘　　要：目的:基于网络药理学和实验验证探讨葛花通过调控细胞自噬干预肝癌的作用机制。方法:通过细胞增殖、划痕实验探索葛花含药血清对HepG2细胞的影响。使用TCMSP数据库获取葛花靶点,从OMIM等数据库获得肝癌疾病靶点,从HADb数据库获得细胞自噬基因,三者取交集后进行蛋白互作(PPI)、GO、KEGG富集分析,并筛选出Degree前15靶点。对筛选出的15个靶点进行差异表达、ROC曲线、临床相关性、生存预后、分子对接和分子动力学模拟分析。对最终筛选出来的靶点进行Western Blot验证,并检测自噬标志物LC3B的表达变化。结果:葛花含药血清可以显著抑制HepG2细胞的增殖和迁移。葛花抗肝癌的最重要靶点是Casp3,Casp3可以和葛花活性成分槲皮素(quercetin)稳定结合。与HepG2对照组相比,葛花含药血清组可显著下调Casp3蛋白表达,上调自噬标志物LC3BⅡ/LC3BⅠ。结论:葛花可通过作用于Casp3调控细胞自噬而干预肝癌。Objective:To investigate the mechanism of intervention of Pueraria lobata in hepatocellular carcinoma by cellular autophagy based on network pharmacology and experimental validation.Methods:The effects of medicated serum of Pueraria lobata on HepG2 cells were explored by cell proliferation and scratch experiments.The TCMSP database was used to obtain the target of Pueraria lobata,the target of liver cancer disease from OMIM and other databases,and the autophagy gene of cells from the HADb database,and the protein interaction,GO and KEGG enrichment analysis were carried out after the intersection of the three targets,and the top 15 targets of Degree were screened.The 15 screened targets were analyzed by differential expression,ROC curves,clinical relevance,survival prognosis,molecular docking and molecular dynamics simulation.The final screened targets were verified by Western blot,and the expression level of LC3B in different doses was detected.Results:Pueraria lobata medicated serum could significantly inhibit the proliferation and migration of HepG2 cells.The most important target of Pueraria lobata against liver cancer was Casp3,which can be stably bound to quercetin,the active ingredient of Pueraria lobata.Compared with the HepG2 control group,the expression of Casp3 protein in the Pueraria lobata sera group was significantly down-regulated,and LC3BII/LC3BI was significantly increased.Conclusion:Pueraria lobata can intervene in liver cancer via Casp3 mediated cellular autophagy.

关 键 词：肝癌 葛花 细胞自噬 Casp3 

分 类 号：R285[医药卫生—中药学]