摘要
[目的]观察机械牵张力(SS)是否通过激活PKCα诱导小鼠血管平滑肌细胞(VSMC)增殖和凋亡,并探讨盐酸小檗碱(BBR)对它的作用及机制。[方法]实验共分6组:正常对照组、BBR组、PKCα抑制剂(Go6976)组、SS组、SS+BBR组、SS+Go6976组。静息培养的细胞给予BBR或Go6976预处理1 h,继而予10%强度的SS刺激15 min后收集细胞,Western blot检测PKCα和MAPK(ERK、JNK、p38)磷酸化水平。BBR或Go6976预处理1 h,牵拉时间为1 h,继续培养23 h,免疫荧光法检测细胞增殖(Ki67)和凋亡(TUNEL)。[结果]Western blot与免疫荧光结果显示,与正常对照组相比较,SS可升高PKCα和MAPK(ERK、JNK和p38)磷酸化水平(P<0.05),并增加VSMC的增殖和凋亡水平(P<0.05)。BBR可抑制PKCα和ERK、JNK、p38磷酸化水平(P<0.05),同时抑制VSMC增殖和凋亡(P<0.05);Go6976可抑制PKCα和ERK、JNK磷酸化水平(P<0.05),但对p38磷酸化的增加没有影响,同时抑制VSMC增殖和凋亡(P<0.05)。[结论]BBR通过抑制SS诱导的VSMC的PKCα和MAPK磷酸化,抑制VSMC增殖和凋亡。
Aim To observe whether the proliferation and apoptosis of mouse vascular smooth muscle cells(VSMC)were induced by stretch stress(SS)via PKCαphosphorylation and to investigate the role of berberine(BBR)and related mechanism on the process.Methods The experiment was divided into six groups:normal control(NC)group,BBR group,PKCαinhibitor(Go6976)group,SS group,SS+BBR group and SS+Go6976 group.VSMC were pretreated with BBR or Go6976 for 1 h,and stimulated for 15 min with SS at 10%amplitude.PKCαand MAPK(ERK,JNK,p38)phosphorylation were measured by Western blot.After pretreated by BBR or Go6976,and SS stimulation for 1 h,the proliferation(Ki67)and apoptosis(TUNEL)of VSMC were measured by immunofluorescence.Results Compared with the NC group,SS could increase the phosphorylation levels of PKCαand MAPK(ERK,JNK,p38)(P<0.05),and increase the levels of VSMC proliferation and apoptosis(P<0.05).BBR could inhibit the phosphorylation levels of PKCαand MAPK(P<0.05),while inhibiting VSMC proliferation and apoptosis(P<0.05);Go6976 could inhibit PKCα,ERK and JNK phosphorylation(P<0.05),but had no effect on the phosphorylation increase of p38,while inhibiting VSMC proliferation and apoptosis(P<0.05).Conclusion BBR inhibits the phosphorylation of PKCαand MAPK(ERK,JNK,p38)induced by SS,and further inhibits the proliferation and apoptosis of VSMC.
作者
蔡晓东
刘树迎
林宁
杨浩澜
李朝红
CAI Xiaodong;LIU Shuying;LIN Ning;YANG Haolan;LI Chaohong(Department of Histology and Embryology,Zhongshan Medical College,Sun Yat-sen University,Guangzhou,Guangdong 510080,China)
出处
《中国动脉硬化杂志》
CAS
2023年第6期466-472,共7页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金项目(81870219、81500337)
广东省自然科学基金项目(2017A030313574)
中央高校基本科研业务费专项资金项目(19ykpy171)。
作者简介
蔡晓东,硕士研究生,研究方向为心血管重构分子机制与防治,E-mail:514104360@qq.com;通信作者:李朝红,博士,教授,博士研究生导师,研究方向为心血管重构分子机制与防治,E-mail:lichaoh@mail.sysu.edu.cn。
