Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, 835 S. Wolcott Ave, M/C 901, Chicago, IL 60612-7342, USA.
J Mol Cell Cardiol. 2011 Dec;51(6):988-96. doi: 10.1016/j.yjmcc.2011.09.016. Epub 2011 Sep 24.
Earlier investigations in our lab indicated an anti-adrenergic effect induced by activation of p21-activated kinase (Pak-1) and protein phosphatase 2A (PP2A). Our objective was to test the hypothesis that Pak-1/PP2A is a signaling cascade controlling stress-induced cardiac growth. We determined the effects of ablation of the Pak-1 gene on the response of the myocardium to chronic stress of isoproterenol (ISO) administration. Wild-type (WT) and Pak-1-knockout (Pak-1-KO) mice were randomized into six groups to receive either ISO, saline (CTRL), or ISO and FR180204, a selective inhibitor of Erk1/2. Echocardiography revealed that hearts of the Pak-1-KO/ISO group had increased LV fractional shortening, reduced LV chamber volume in diastole and systole, increased cardiac hypertrophy, and enhanced transmitral early filling deceleration time, compared to all other groups. The changes were associated with an increase in relative Erk1/2 activation in Pak-1-KO/ISO mice versus all other groups. ISO-induced cardiac hypertrophy and Erk1/2 activation in Pak-1-KO/ISO were attenuated when the selective Erk1/2 inhibitor FR180204 was administered. Immunoprecipitation showed an association between Pak-1, PP2A, and Erk1/2. Cardiac myocytes infected with an adenoviral vector expressing constitutively active Pak-1 showed a repression of Erk1/2 activation. p38 MAPK phosphorylation was decreased in Pak-1-KO/ISO and Pak-1-KO/CTRL mice compared to WT. Levels of phosphorylated PP2A were increased in ISO-treated Pak-1-KO mice, indicating reduced phosphatase activity. Maximum Ca(2+)-activated tension in detergent-extracted bundles of papillary fibers from ISO-treated Pak-1-KO mice was higher than in all other groups. Analysis of cTnI phosphorylation indicated that compared to WT, ISO-induced phosphorylation of cTnI was blunted in Pak-1-KO mice. Active Pak-1 is a natural inhibitor of Erk1/2 and a novel anti-hypertrophic signaling molecule upstream of PP2A.
早期我们实验室的研究表明,p21 激活激酶(Pak-1)和蛋白磷酸酶 2A(PP2A)的激活可诱导抗肾上腺素能作用。我们的目的是验证 Pak-1/PP2A 是否是控制应激诱导的心脏生长的信号级联假说。我们确定了 Pak-1 基因缺失对异丙肾上腺素(ISO)给药引起的心肌对慢性应激反应的影响。野生型(WT)和 Pak-1 敲除(Pak-1-KO)小鼠被随机分为六组,分别接受 ISO、生理盐水(CTRL)或 ISO 和选择性 Erk1/2 抑制剂 FR180204。超声心动图显示,与所有其他组相比,Pak-1-KO/ISO 组的左心室(LV)分数缩短增加,LV 腔舒张和收缩容积减小,心脏肥大增加,E 峰减速时间加快。这些变化与 Pak-1-KO/ISO 小鼠中相对 Erk1/2 激活增加有关。当给予选择性 Erk1/2 抑制剂 FR180204 时,ISO 诱导的 Pak-1-KO/ISO 心脏肥大和 Erk1/2 激活减弱。免疫沉淀显示 Pak-1、PP2A 和 Erk1/2 之间存在关联。用表达组成型激活的 Pak-1 的腺病毒载体感染心肌细胞后,Erk1/2 激活受到抑制。与 WT 相比,Pak-1-KO/ISO 和 Pak-1-KO/CTRL 小鼠的 p38 MAPK 磷酸化减少。ISO 处理的 Pak-1-KO 小鼠中磷酸化的 PP2A 水平增加,表明磷酸酶活性降低。从 ISO 处理的 Pak-1-KO 小鼠的心肌细胞中提取的肌球蛋白纤维束中,最大钙激活张力高于所有其他组。肌钙蛋白 I 磷酸化分析表明,与 WT 相比,ISO 诱导的 Pak-1-KO 小鼠 cTnI 磷酸化减弱。活性 Pak-1 是 Erk1/2 的天然抑制剂,也是 PP2A 上游的新型抗肥大信号分子。