基于网络药理学及实验验证探讨雷公藤红素抗脑胶质瘤的作用机制  

Study on anti-glioma mechanism of celastrol based on network pharmacology and experimental verification

作　　者：杜晓丹 范孟杨 徐柳清 赵培源[1] 杜志欣 刘征 侯俊林[1] 刘喜红[1] 杨丽萍[1] DU Xiao-dan;FAN Meng-yang;XU Liu-qing;ZHAO Pei-yuan;DU Zhi-xin;LIU Zheng;HOU Jun-lin;LIU Xi-hong;YANG Li-ping(Traditional Chinese Medicine(Zhong Jing)School,Henan University of Chinese Medicine,Zhengzhou HENAN 450046,China)

机构地区：[1]河南中医药大学中医学院(仲景学院),河南郑州450046

出　　处：《中国新药与临床杂志》2025年第3期213-220,共8页Chinese Journal of New Drugs and Clinical Remedies

基　　金：国家自然科学基金青年科学基金项目(82004108);河南省科技攻关项目(202102310385);2023年度河南省博士后科研资助项目(HN2024094);河南省高等学校青年骨干教师培养计划(2023GGJS082)。

摘　　要：目的通过网络药理学结合细胞实验探究雷公藤红素(Cel)抗脑胶质瘤细胞生长的分子机制。方法利用TCMSP和Swiss Target Prediction数据库获取Cel药物靶点,GeneCards和TTD数据库获取脑胶质瘤疾病靶点,将两者共有靶点导入STRING数据库构建蛋白质相互作用(PPI)网络,利用Cytoscape 3.7.2软件进行可视化,并进行GO和KEGG富集分析。采用GEPIA软件筛选出与脑胶质瘤生存密切相关的靶点,通过Autodock软件进行分子对接和可视化。采用CCK-8法和克隆形成实验评估Cel对脑胶质瘤细胞增殖能力,Western blot法检测核心靶点及信号通路相关蛋白表达。结果筛选出Cel与脑胶质瘤的共有靶点87个,主要涉及PI3K/Akt、VEGF等信号通路。其中VEGFA和FN1在脑胶质瘤组织中的表达明显高于正常脑组织(P<0.01),且两者的表达水平与患者的生存期呈负相关,Cel与FN1、VEGFA的结合能为-8.3、-7.3 kcal·mol^(-1)。体外细胞实验结果显示,与空白组相比,Cel各浓度组细胞增殖能力显著降低(P<0.01),VEGFA、FN1、p-PI3K和p-Akt蛋白表达显著下调(P<0.05)。结论Cel可能通过抑制PI3K/Akt信号通路,降低VEGFA和FN1的表达,抑制脑胶质细胞的增殖。AIM To investigate the molecular mechanism of celastrol(Cel)anti-glioma cell growth by combining network pharmacology and cellular experiment.METHODS The targets for the Cel drug were obtained from the TCMSP and Swiss Target Prediction databases.Glioma disease targets were collected from the GeneCards and TTD databases.The common targets were then inputted into the STRING database to construct a protein-protein interaction(PPI)network,visualized via Cytoscape 3.7.2 software.Subsequently,GO and KEGG analysis of intersected genes.GEPIA software was used to screen out the targets that were closely related to the survival of gliomas,and molecular docking and visualization were performed by Autodock software.The CCK-8 assay and clone formation were used to assess the cell proliferation ability of Cel on giloma cells,and Western blot was used to verify the expressions of core targets and signaling pathway-related proteins.RESULTS Eighty-seven targets shared by Cel and glioma were screened,mainly involving PI3K/Akt,VEGF and other signaling pathways.The expressions of VEGFA and FN1 in glioma tissues were significantly higher than that in normal brain tissues(P<0.01),and the expressions of the two were negatively correlated with the survival of the patients,and the binding energies of Cel with FN1 and VEGFA were-8.3 and-7.3 kcal·mol^(-1).In vitro cellular experiments demonstrated a notable reduction in cell proliferation ability in the Cel group compared to the blank group(P<0.01).Furthermore,protein expressions of VEGFA,FN1,p-PI3K,and p-Akt were significantly down-regulated(P<0.05).CONCLUSION Cel may inhibit the proliferation of U251 cells by inhibiting the PI3K/Akt signaling pathway and reducing the expressions of VEGFA and FN1.

关 键 词：雷公藤红素 脑胶质瘤 网络药理学 分子对接 生存分析 

分 类 号：R273[医药卫生—中西医结合]