基于TLR4/MyD88/NF-κB信号通路探讨豆甾醇抗肝细胞癌的作用机制  

Study on mechanism of stigmasterol inhibiting hepatocellular carcinoma based on TLR4/MyD88/NF-κB signaling pathway

作　　者：于春 马燕花[1] YU Chun;MA Yan-hua(The First Clinical Medicine School,Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学第一临床医学院,甘肃兰州730000

出　　处：《中国药理学通报》2025年第5期867-873,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金地区基金项目(No 81860821);甘肃省联合科研基金一般项目(No 23JRRA1523)。

摘　　要：目的采用分子对接和体外实验,探讨豆甾醇(stigmasterol)对肝细胞癌(hepatocellular carcinoma,HCC)中TLR4/MyD88/NF-κB信号通路的调节机制。方法使用AutoDock程序进行分子对接,研究豆甾醇与Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(myeloid differentiation primary response gene 88,MyD88)和核因子-κB(nuclear factor-κB,NF-κB)的结合亲和力。通过CCK-8、愈合实验和Transwell实验评估其对细胞增殖和迁移的影响,同时通过克隆形成实验测定克隆形成能力。利用流式细胞术检查细胞凋亡和细胞周期。Western blot分析用于评估豆甾醇处理后的TLR4、MyD88和NF-κB蛋白表达情况。结果分子对接显示豆甾醇与TLR4、MyD88和NF-κB具有良好的结合构象。豆甾醇在体外对肝细胞癌细胞的增殖、迁移、侵袭和克隆形成能力表现出抑制作用。此外,它促进了细胞凋亡,抑制了细胞周期,并减少了TLR4/MyD88/NF-κB信号通路的蛋白表达(P<0.05)。结论豆甾醇通过调节TLR4/MyD88/NF-κB信号通路抑制了肝细胞癌细胞的活性。Aim To utilize molecular docking and in vitro experiments to investigate the regulatory mechanism of stigmasterol on the TLR4/MyD88/NF-κB signaling pathway in hepatocellular carcinoma.Methods Molecular docking was performed using AutoDock to study the binding affinity of stigmasterol with TLR4,MyD88,and NF-κB.The effects on cell proliferation and migration were assessed through CCK-8,wound healing assays,and Transwell experiments,while colony formation ability was measured using a colony formation assay.Flow cytometry was employed to examine apoptosis and cell cycle.Western blot analysis was used to evaluate the expression levels of TLR4,MyD88,and NF-κB proteins after stigmasterol treatment.Results Molecular docking indicated that stigmasterol exhibited favorable binding conformations with TLR4,MyD88,and NF-κB.In vitro,stigmasterol demonstrated inhibitory effects on proliferation,migration,invasion,and colony formation in hepatocellular carcinoma cells.Additionally,it promoted apoptosis,inhibited cell cycle progression,and reduced protein expression of the TLR4/MyD88/NF-κB signaling pathway(P<0.05).Conclusion Stigmasterol suppresses the activity of hepatocellular carcinoma cells by modulating the TLR4/MyD88/NF-κB signaling pathway.

关 键 词：豆甾醇 TOLL样受体4 髓样分化因子88 核因子-ΚB 肝细胞癌 凋亡 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学] R329.28R392.11R735.702.2