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A Novel RhoA/ROCK-CPI-17-MEF2C Signaling Pathway Regulates Vascular Smooth Muscle Cell Gene Expression

机译:新型RhoA / ROCK-CPI-17-MEF2C信号通路调节血管平滑肌细胞基因表达

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摘要

Differentiation of vascular smooth muscle cells (VSMC) is a fundamental aspect of normal development and vascular disease. During contraction, VSMCs modulate calcium sensitivity through RhoA/ROCK-mediated inhibition of the myosin light chain phosphatase complex (MLCP). Previous studies have demonstrated that this signaling pathway functions in parallel to increase the expression of smooth muscle genes through the myocardin-family of co-activators. MEF2C fulfills a critical role in VSMC differentiation and regulates myocardin expression, leading us to investigate whether the RhoA/ROCK signaling cascade might regulate MEF2 activity. Depolarization-induced calcium signaling increased the expression of myocardin, which was sensitive to ROCK and p38 MAPK inhibition. We previously identified protein phosphatase 1α (PP1α), a known catalytic subunit of the MLCP in VSMCs, as a potent repressor of MEF2 activity. PP1α inhibition resulted in increased expression of myocardin, while ectopic expression of PP1α inhibited the induction of myocardin by MEF2C. Consistent with these data, shRNA-mediated suppression of a PP1α inhibitor, CPI-17, reduced myocardin expression and inhibited VSMC differentiation, suggesting a pivotal role for CPI-17 in regulating MEF2 activity. These data constitute evidence of a novel signaling cascade that links RhoA-mediated calcium sensitivity to MEF2-dependent myocardin expression in VSMCs through a mechanism involving p38 MAPK, PP1α, and CPI-17.
机译:血管平滑肌细胞(VSMC)的分化是正常发育和血管疾病的基本方面。在收缩过程中,VSMC通过RhoA / ROCK介导的肌球蛋白轻链磷酸酶复合物(MLCP)抑制作用来调节钙敏感性。先前的研究表明,该信号传导途径平行发挥作用,以通过共激活因子的心肌家族增加平滑肌基因的表达。 MEF2C在VSMC分化中起关键作用,并调节心肌表达,从而使我们研究RhoA / ROCK信号级联反应是否可能调节MEF2活性。去极化诱导的钙信号传导增加了心肌蛋白的表达,其对ROCK和p38 MAPK抑制敏感。我们之前鉴定出蛋白磷酸酶1α(PP1α)是VSMC中MLCP的已知催化亚基,可作为MEF2活性的有效阻遏物。 PP1α抑制导致心肌蛋白表达增加,而异位表达PP1α抑制MEF2C诱导心肌蛋白。与这些数据一致,shRNA介导的PP1α抑制剂CPI-17的抑制可降低心肌素表达并抑制VSMC分化,提示CPI-17在调节MEF2活性中起关键作用。这些数据构成了一种新颖的信号级联的证据,该信号级联通过涉及p38 MAPK,PP1α和CPI-17的机制将RhoA介导的钙敏感性与VSMC中MEF2依赖的心肌表达联系起来。

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