黄连素抑制机械牵张力诱导的血管平滑肌细胞MAPK磷酸化以及细胞增殖和迁移  

Berberine Inhibits Mechanical Stretch Stress induced MAPK Phosphorylation,Proliferation and Migration of Vascular Smooth Muscle Cells

作　　者：邓烈 王林丽 刘树迎[1] 李朝红[1] DENG Lie;WANG Lin-li;LIU Shu-ying;LI Chao-hong(Department of Histology and Embryology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学中山医学院组织学与胚胎学教研室

出　　处：《中山大学学报（医学版）》2020年第1期44-52,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81070124;81870219;81500337);广东省自然科学基金(2017A030313574;2014A020212109)

摘　　要：[目的]观察黄连素能否阻断机械牵张力引起的血管平滑肌细胞(VSMC)增殖,并探讨MAPK信号通路在其中的作用。[方法]实验分4组:NC组、SS组、BBR+SS组以及BBR组。NC组给予PBS处理;SS组为牵拉组,给予牵张力刺激;BBR+SS组为黄连素干预组,先用一定浓度的黄连素对VSMC预处理1h后再给予牵张力刺激;BBR组的VSMC只用黄连素刺激1h,之后换无血清培养继续培养。然后分别收集VSMC,分别采用Westernblot方法分析VSMC的MAPK磷酸化水平变化,采用免疫荧光检测VSMC的增殖情况,以及采用划痕实验方法检测VSMC的迁移情况。[结果]与NS组相比,SS可明显引起VSMC增殖与迁移增加,黄连素则可明显抑制SS引起的VSMC增殖与迁移增加。机制上,机械牵张力作用可引起VSMC的MAPK(ERK,JNK,P38)磷酸化明显增加,而黄连素呈浓度依赖性抑制SS诱导的MAPK磷酸化增加。[结论]黄连素可通过MAPK通路抑制高血压机械力诱导的血管平滑肌细胞增殖、迁移。该研究结果揭示了黄连素旧药新用途、新机制,为临床上深入研究高血压机械力异常增加引起的血管重构防治机制提供重要资料。【Objective】To observe whether berberine can inhibit vascular smooth muscle cells(VSMC)proliferation induced by mechanical strength stress and to investigate the role of MAPK pathway in it.【Methods】The cultured VSMC were divided into 4 groups:negative control group(NC group),stretch stress group(SS group),berberine pretreated and stretch stress stimulation group(BBR+SS group),and breberinegroup.In NC group,phosphate buffer saline was used as a negative control;in SS group,stretch stress was given to VSMC;in BBR+SS group,VSMC were pretreated with berber⁃ine for 1 hour and then exposed to stretch stress;in BBR group,VSMC were treated only with berberine for 1 hour and cultured in serum-free DMEM afterwards.We collected VSMC in each group,detected and analyzed their MAPK phos⁃phorylation,proliferation and migration by using Western blotting,immunofluorescence and wound-healing assay respec⁃tively.【Results】Compare with NC group,stretch stress markedly induced VSMC proliferation and migration,which could be inhibited significantly by berberine.Stretch stress obviously increased phosphorylation of MAPK(ERK,JNK,p38),which could be inhibited by berberine in a concentration dependent manner.【Conclusion】Berberine inhibits hypertension-induced proliferation and migration of VSMC through MAPK pathway.The results revealed the new use and mechanism of berberine,and provided important data for further study on the prevention and treatment of vascular remod⁃eling caused by abnormal increase of mechanical stress in hypertension.

关 键 词：血管平滑肌细胞 机械牵张力 黄连素 细胞增殖/迁移 MAPK磷酸化 高血压 

分 类 号：R63[医药卫生—外科学]