作　　者： ; ; ; ; ;

机构地区： 北京林业大学

出　　处： 《中国生物化学与分子生物学报》 2011年第11期1039-1043,共5页

摘　　要： Erk1/2活性在血管许多细胞功能中具有重要影响,而Notch3主要表达在动脉平滑肌细胞中,并且是发育过程中动脉成熟所必需的.为了探讨Notch3在血管平滑肌细胞中对Erk1/2信号通路的调控作用,采用siRNA基因敲除Notch3,γ-分泌酶抑制剂DAPT抑制Notch信号通路,质粒转染过表达Notch3活性区等方法,用Western印迹检测Notch3对血管平滑肌细胞中Erk1/2磷酸化水平,即Erk1/2活性的影响.同时,利用活性氧自由基(ROS)诱导激活Erk1/2;siRNA敲除Notch3表达致使血管平滑肌细胞中Erk1/2的磷酸化水平显著降低,并且抑制了ROS诱导的Erk1/2激活;同样,Notch通路抑制剂DAPT也抑制了ROS诱导的Erk1/2激活;而Notch3活性区NICD的过表达并没有改变血管平滑肌细胞中Erk1/2的磷酸化水平,但其延缓了ROS激活后Erk1/2活性的衰减.上述结果表明,Notch3可在血管平滑肌细胞中调控Erk1/2活性以及ROS诱导的Erk1/2信号激活. Activation of Erk1/2 affects a broad range of cellular functions in blood vessels.Notch3 is mainly expressed in vascular smooth muscle cells of arteries,and is necessary for arterial maturity during the development.In order to investigate the impact of Notch3 on Erk1/2 activation in vascular smooth muscle cells,Erk1/2 activation was induced by reactive oxygen species（ROS） and the level of Erk1/2 phosphorylation was analyzed by Western blotting after transfection with Notch3 siRNA or construct with Notch3 intracellular domain（NICD）,or blocking Notch signaling by γ-secretase inhibitor DAPT.Erk1/2 phosphorylation was decreased in vascular smooth muscle cells after siRNA knockout of Notch3 expression.Both Notch3 siRNA knockdown and DAPT inhibited ROS-induced Erk1/2 activation.No change in Erk1/2 phosphorylation was detected in NICD overexpressed vascular smooth muscle cells.However,NICD overexpression postponed the attenuation of ROS-induced Erk1/2 activation.In conclusion,Notch3 can regulate Erk1/2 activity and ROS-induced Erk1/2 activation in vascular smooth muscle cells.

关 键 词： 血管平滑肌细胞 活性氧自由基

领　　域： [生物学]