基于网络药理学探讨芪珠消癥方抗肝细胞癌的机制及PI3K/Akt途径验证  

Mechanism of Qizhu Xiaozheng Formula Against Hepatocellular Carcinoma Based on Network Pharmacology and the Verification of PI3K/Akt Pathway

作　　者：吴驻林 魏春山[1] 谭婉君 刘林华 王李安安 何力[1] 彭立生[1] WU Zhu-lin;WEI Chun-shan;TAN Wan-jun;LIU Lin-hua;WANG li-an-an;HE Li;PENG li-sheng(The Fourth Clinical Medical Cdllege of Guangzhou University of Chinese Medicine(Shenzhen Traditional Chinese Medicine Hospital),Shenzhen Guangdong 518000;Shenzhen Futian Center for Chronie Disease Contrdl,Shenzhen Guangdong 518000;The University of Hong Kong-Shenzhen Hospital,Shenzhen Guangdong 518000)

机构地区：[1]广州中医药大学第四临床医学院(深圳市中医院),广东深圳518000 [2]深圳市福田区慢性病防治院,广东深圳518000 [3]香港大学深圳医院,广东深圳518000

出　　处：《世界中西医结合杂志》2023年第1期1-11,67,共12页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(82074384);深圳市科技计划项目(JCYJ20210324111207020,JCYJ20190812164211151)。

摘　　要：目的探讨芪珠消癥方治疗肝细胞癌(Hepatocellular Carcinoma,HCC)的潜在作用机制。方法分别通过5个中药数据库与2个疾病数据库筛选芪珠消癥方活性成分、作用靶点与HCC疾病靶点,运用网络药理学技术构建芪珠消癥方与HCC关系网络,并通过蛋白互作网络、通路富集分析、分子对接预测机制。采用实验验证芪珠消癥方含药血清及其关键成分槲皮素(Quercetin,QCT)对HCC干预作用,并检测QCT对磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)途径的影响。结果芪珠消癥方与HCC关系网络提示芪珠消癥方主要通过QCT、汉黄芩素等关键活性成分调控HCC的相关靶点。蛋白互作网络、通路富集分析显示芪珠消癥方可能通过AKT1、丝裂原活化蛋白激酶3(MAPK3)等关键靶点与PI3K/Akt、乙型肝炎、凋亡等重要信号途径发挥抗HCC作用。分子对接显示AKT1蛋白与QCT亲和力较好;实验结果表明,芪珠消癥方含药血清可有效地抑制HepG2细胞增殖并促进其凋亡,关键活性成分QCT以浓度相关的方式抑制HepG2细胞的增殖、克隆形成、迁移能力,促使细胞凋亡,并可降低磷酸化蛋白p-PI3K、p-Akt的表达。结论芪珠消癥方治疗HCC的主要机制包括以QCT、汉黄芩素为代表的多活性成分,以PI3K/Akt、乙型肝炎、凋亡代表的多途径的协同调节作用。Objective To explore the potential mechanism of Qizhu Xiaozheng(QZXZ)formula in the treatment of hepatocellular carcinoma(HCC).Methods The bioactive ingredients and targets of QZXZ formula and HCC targets were screened out by five Chinese medicine databases and two disease databases,respectively.The network pharmacology was utilized to construct the relationship network between QZXZ formula and HCC,and the mechanism was predicted by protein-protein interaction(PPI)network,pathway enrichment analysis,and molecular docking.Experiments were performed to verify the intervention effect of QZXZ formula-containing serum and its key ingredient[quercetin(QCT)]on HCC,and to detect the effect of QCT on phosphatidylinositol 3-kinases/protein kinase B(PI3K/Akt)pathway.Results The relationship network between QZXZ formula and HCC suggested that QZXZ formula mainly regulated the related targets of HCC through key bioactive ingredients such as QCT and wogonin.PPI network and pathway analyses showed that QZXZ formula may play an anti-HCC effect through key targets such as protein kinase B1(AKT1),mitogen-activated protein kinase 3(MAPK3)and important signaling pathways such as PI3K-Akt,hepatitis B,and apoptosis.Molecular docking displayed that AKT1 protein had a good affinity with QCT.The experimental results showed that QZXZ formula-containing serum could effectively inhibit the proliferation and promote the apoptosis of HepG2 cells.QCT,the key bioactive ingredients,could inhibit the proliferation,clone formation and migration of HepG2 cells in a concentration-dependent behavior,promote HepG2 cells apoptosis,and decrease the expression of phosphorylated proteins p-PI3K and p-Akt.Conclusion The main mechanism of QZXZ formula in the treatment of HCC includes multiple bioactive ingredients representedby QCT and wogonin and multi-pathways represented by PI3K-Akt,hepatitis B,and apoptosis.

关 键 词：芪珠消癥方 叶下珠 肝癌 网络药理学 槲皮素 

分 类 号：R285.6[医药卫生—中药学]