基于网络药理学和血清药理学探讨海牡方治疗非小细胞肺癌的作用机制

Exploring the mechanism of Haimu Decoction in the treatment of non-small cell lung cancer based on network pharmacology and serum pharmacology

关键词:  网络药理学  海牡方  含药血清  细胞周期  细胞凋亡

Key words:  network pharmacology  Haimufang decoction (HMF)  medicated serum  cell cycle  cell apoptosis

摘要:

摘要：目的 通过运用网络药理学和血清药理学方法探讨海牡方（HMF）治疗非小细胞肺癌（NSCLC）的主要成分、核心靶点、关键信号通路以阐明其可能的作用机制。方法 分别采用TCMSP和DisGeNET等据库检索、汇总HMF的化合物成分、作用靶点及非小细胞肺癌疾病相关靶点；将药物的作用靶点与疾病的靶点取交集获得共有靶点，并上传至 STRING 数据平台构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络筛选核心靶点；运用DAVID数据库对核心靶点进行GO及KEGG富集分析，并采用Cytoscape 3.9.1软件构建HMF“药味-成分-靶点-通路”网络，预测HMF可能的作用机制；最后通过HMF含药血清对非小细胞肺癌NCI-H1975细胞的细胞周期阻滞和细胞凋亡诱导实验验证HMF对肺癌的作用及其潜在机制。结果 从HMF中鉴定了65个化合物成分，筛选出24-Keto-Cholesterol，quercetin，Diisobutyl phthalate等47个主要化合物成分，PPI网络分析结果显示，肿瘤蛋白p53（tumor protein p53,TP53）、肿瘤坏死因子（tumor necrosis factor，TNF）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）等共55个可能是HMF治疗NSCLC的核心靶蛋白，涉及癌症通路（Pathways in cancer）等多条信号通路。在随后的验证实验中，Pathways in cancer通路中的细胞周期阻滞和凋亡诱导已被证实参与抑制非小细胞肺癌。结论 HMF通过多成分、多靶点和多途径信号通路的方式发挥治疗非小细胞肺癌的作用，为HMF的作用机制和临床应用提供了理论基础。

Abstract:

ABSTRACT: OBJECTIVE To elucidate the possible mechanism, the network pharmacology and serum pharmacology approaches were performed to explore the main components, core targets, and key signaling pathways of Haimu Decoction (HMF) in treating non-small cell lung cancer (NSCLC). METHODS TCMSP and DisGeNET databases were used to search and summarize the components, targets and non-small cell lung cancer-related targets of HMF, respectively. The common targets were obtained by intersection of drug targets and disease targets, and uploaded to the STRING data platform to construct a protein-protein interaction (PPI) network to screen core targets. GO and KEGG enrichment analysis of core targets were performed using DAVID database. Cytoscape 3.9.1 software was used to construct the network of“HMF flavor-components-targets-pathways”. Finally, the cell cycle arrest and apoptosis induction experiments of HMF-containing serum on non-small cell lung cancer NCI-H1975 cells were used to verify the effect of HMF on lung cancer and its potential mechanism. RESULTS A total of 65 compounds were identified from HMF, and 47 main compounds such as 24-Keto-Cholesterol, quercetin and Diisobutyl phthalate were screened out. PPI network analysis shown that tumor protein p53 (TP53), tumor necrosis factor (TNF), cystein-asparate protease-3 (Caspase-3) and other 55 proteins might be the core target proteins of HMF in the treatment of NSCLC, involving multiple signaling pathways such as pathways in cancer. In subsequent validation experiments, cell cycle arrest and apoptosis induction in the Pathways in cancer pathway have been shown to be involved in the inhibition of non-small cell lung cancer. CONCLUSION HMF plays a role in the treatment of non-small cell lung cancer through multi-component, multi-target and multi-channel signaling pathways, which provides a theoretical basis for the mechanism and clinical application of HMF.