Heat stress activates AKT via focal adhesion kinase-mediated pathway in neonatal rat ventricular myocytes.
Heat stress activates AKT via focal adhesion kinase-mediated pathway in neonatal rat ventricular myocytes.
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热应激通过新生大鼠心室肌细胞中粘着斑激酶介导的途径激活 AKT。
DOI：
10.1152/ajpheart.00401.2008
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
VanderHeide,RichardS
中科院分区：
文献类型：
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作者：
Wei,Hongguang;VanderHeide,RichardS
热应激（HS）诱导的心脏保护作用与桩蛋白在新生大鼠心室肌细胞（NRVM）膜部分的定位增加有关。本研究的目的是1）检测HS激活的亚细胞信号通路;2）确定心肌应激是否组织和激活整合的存活通路; 3）研究HS激活的潜在下游细胞保护蛋白。在HS之后，NRVM经受化学抑制剂（CI），该化学抑制剂被设计成通过抑制糖酵解和线粒体呼吸来模拟缺血。用腺病毒载体选择性地增加蛋白激酶B（AKT）表达（野生型）。用Western印迹分析分析细胞信号传导，同时分别通过测量台盼蓝排斥和/或末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色来测定胀亡/凋亡。HS增加了粘着斑激酶（FAK）在酪氨酸397的磷酸化，但没有不利地影响CI前NRVM的活力。HS增加FAK和磷脂酰肌醇3-激酶之间的关联，以及引起AKT活性的显着增加。野生型AKT表达的增加保护了心肌细胞免于细胞凋亡和细胞凋亡。与表达空病毒的心肌细胞相比，FAK抑制剂FRNK的表达增加，在时间0和CI 10分钟后对HS的反应中减少AKT磷酸化。我们的结论是，心肌应激激活与保护致死性细胞损伤相关的基于细胞因子的信号通路。
Heat stress (HS)-induced cardioprotection is associated with increased paxillin localization to the membrane fraction of neonatal rat ventricular myocytes (NRVM). The purpose of this study was1) to examine the subcellular signaling pathways activated by HS;2) to determine whether myocardial stress organizes and activates an integrated survival pathway; and3) to investigate potential downstream cytoprotective proteins activated by HS. After HS, NRVM were subjected to chemical inhibitors (CI) designed to simulate ischemia by inhibiting both glycolysis and mitochondrial respiration. Protein kinase B (AKT) expression (wild type) was increased selectively with an adenoviral vector. Cell signaling was analyzed with Western blot analysis, while oncosis/apoptosis was assayed by measuring Trypan blue exclusion and/or terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining, respectively. HS increased phosphorylation of focal adhesion kinase (FAK) at tyrosine 397 but did not adversely affect the viability of NRVM before CI. HS increased association between FAK and phosphatidylinositol 3-kinase as well as causing a significant increase in AKT activity. Increased expression of wild-type AKT protected myocytes from both oncotic and apoptotic cell death. Increased expression of a FAK inhibitor, FRNK, reduced AKT phosphorylation in response to HS both attime 0and after 10 min of CI compared with myocytes expressing empty virus. We conclude that myocardial stress activates cytoskeleton-based signaling pathways that are associated with protection from lethal cell injury.