基于网络药理学和实验验证探究莪术醇对肝星状细胞衰老的作用机制  

Exploring the Mechanism of Action of Curcumol on Hepatic Stellate Cell Senescence Based on Network Pharmacology and Experimental Validation

作　　者：郑洋 王佳慧 赵铁建 王鸿红 汪磊[1,2] ZHENG Yang;WANG Jiahui;ZHAO Tiejian;WANG Honghong;WANG Lei(Department of Medicine,Faculty of Chinese Medicine Science,Guangxi University of Chinese Medicine,Nanning 530222,China;Graduate School,Guangxi University of Chinese Medicine,Nanning 530222,China)

机构地区：[1]广西中医药大学赛恩斯新医药学院医学系,南宁530222 [2]广西中医药大学研究生学院,南宁530222

出　　处：《中国现代应用药学》2024年第23期3355-3365,共11页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金项目(82204755);广西自然科学基金项目(2024GXNSFAA010235、2023GXNSFBA026274);广西壮瑶药重点实验室基金资助项目(GXZYYKF2023-05);广西研究生教育创新计划项目(YCBZ2024150);广西中医药大学赛恩斯新医药学院科研项目(2024ZZA003、2024ZZB007、2024ZZB010、2022MS010)。

摘　　要：目的基于网络药理学和实验验证探究莪术醇对肝星状细胞衰老的分子机制。方法采用Pharm Mapper数据库获取莪术醇作用靶点;在GeneCard、OMIM、PharmGKB数据库收集细胞衰老相关靶点。通过对莪术醇作用靶点和细胞衰老相关靶点进行交互处理得到关键靶点。基于String数据库构建关键靶点互作网络并通过Cytoscape软件进行可视化。对关键靶点进行途径和功能的富集分析,同时将关键靶点与莪术醇进行分子对接分析。借助分子生物学试验检测莪术醇对肝星状细胞衰老的作用及机制。结果共获取莪术醇作用靶点152个,细胞衰老相关靶点2937个,莪术醇调控细胞衰老关键靶点71个。富集分析发现关键靶点主要参与代谢过程、转录因子活性、抗氧化活性等,主要富集在IL-17、VEGF、Ras、自噬、MAPK等信号通路。分子对接结果表明莪术醇与关键靶点具有较好的结合能力。CCK8试验发现莪术醇显著抑制了细胞增殖(P<0.01);分子生物学试验发现莪术醇阻滞细胞周期在G0/G1期(P<0.05);提高细胞衰老与自噬相关蛋白的表达(P<0.05);抑制MAP8和MAPK14 mRNA以及细胞外基质表达(P<0.01);透射电镜检测发现莪术醇显著增加自噬小体数量(P<0.01);β-半乳糖苷酶染色检测发现莪术醇诱导细胞衰老的作用被3-MA挽救。结论莪术醇通过靶向MAPK8和MAPK14调控细胞自噬,诱导肝星状细胞衰老可能是莪术醇防治肝纤维化的关键机制之一。OBJECTIVE To investigate the molecular mechanism of curcumol on hepatic stellate cell senescence based on network pharmacology and experimental validation.METHODS Curcumol targets were obtained from Pharm Mapper database;cellular senescence-related targets were collected from GeneCard,OMIM and PharmGKB databases.The key targets were obtained by interacting curcumol targets and cellular senescence-related targets.The key target interaction network was constructed based on the String database and visualized by Cytoscape software.The key targets were enriched for pathways and functions,and molecular docking analysis was performed between the key targets and curcumol.The effect and mechanism of curcumol on hepatic stellate cell senescence was examined by molecular biology experiments.RESULTS A total of 152 curcumol action targets,2937 cellular senescence-related targets and 71 key targets of curcumol regulation of cellular senescence were obtained.The enrichment analysis revealed that the key targets were mainly involved in metabolic process,transcription factor activity,antioxidant activity,etc.,mainly enriched in IL-17,VEGF,Ras,autophagy,MAPK and other signaling pathways;and the molecular docking results showed that curcumol has better binding ability with the key targets;CCK8 experiment found that curcumol significantly inhibited cell proliferation(P<0.01);molecular biology experiment found that curcumol blocked cell cycle in G0/G1 phase(P<0.05);increased the expression of cellular senescence and autophagy-related proteins(P<0.05);inhibited MAP8 and MAPK14 mRNA and extracellular matrix expression(P<0.01);transmission electron microscopy detected that curcumol significantly increased the number of autophagic vesicles(P<0.01);β-Galactosidase staining assay revealed that curcumol-induced cellular senescence was rescued by 3-MA.CONCLUSION Curcumol regulates cellular autophagy by targeting MAPK8 and MAPK14,and induces hepatic stellate cell senescence may be one of the key mechanisms of curcumol against liver fibrosis.

关 键 词：莪术醇 肝星状细胞 衰老 肝纤维化 网络药理学 自噬 

分 类 号：R285.5[医药卫生—中药学]