Effect of increased expression of cytoskeletal protein vinculin on ischemia-reperfusion injury in ventricular myocytes.
Effect of increased expression of cytoskeletal protein vinculin on ischemia-reperfusion injury in ventricular myocytes.
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DOI：
10.1152/ajpheart.00525.2002
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发表时间：
2003-03
期刊：
影响因子：
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通讯作者：
Hongguang Wei;T. L'ecuyer;R. V. Vander Heide
中科院分区：
文献类型：
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作者：
Hongguang Wei;T. L'ecuyer;R. V. Vander Heide
从可逆性到不可逆性缺血性损伤（缺血-再灌注，I/R）的转变与粘着斑蛋白的损失同时发生，粘着斑蛋白是一种参与肌原纤维附着到肌膜的细胞骨架蛋白。如果黏着斑蛋白的丢失对I/R的发展至关重要，那么假设黏着斑蛋白是功能性的并且定位于适当的亚细胞位点，则可以预测黏着斑蛋白水平的增加会延迟不可逆损伤的发生。本研究确定是否增加表达黏着斑蛋白，特别是在细胞骨架室，将提供保护，从I/R损伤。培养新生大鼠心肌细胞，并感染一种新创建的复制缺陷型腺病毒驱动黏着斑蛋白的表达。糖酵解和线粒体呼吸的化学抑制剂诱导I/R。用乳酸脱氢酶（LDH）释放评估不可逆的细胞损伤。病毒感染的心肌细胞在细胞骨架部分中表达了显著更多的黏着斑蛋白，并增加了桩蛋白的表达，但在对模拟I/R的反应中与对照细胞维持了相同量的损伤（n = 4; P =不显著，配对t检验）。低温I/R（25 ℃缺血）导致LDH释放显著减少（P </= 0.02; n = 4）。病毒介导的黏着斑蛋白过表达似乎并不代表克服I/R损伤的合理方法。
The transition from reversible to irreversible ischemic injury (ischemia-reperfusion, I/R) occurs coincident with the loss of vinculin, a cytoskeletal protein involved in the attachment of the myofibrils to the sarcolemmal membrane. If the loss of vinculin were critical to the development of I/R, then increased levels of vinculin would be predicted to delay the onset of irreversible injury assuming that the protein is functional and localized to the proper subcellular site. The present study determined whether increased expression of vinculin, specifically in the cytoskeletal compartment, would provide protection from I/R injury. Neonatal rat myocytes were cultured and infected with a newly created replication-deficient adenovirus driving the expression of vinculin. I/R was induced with chemical inhibitors of glycolysis and mitochondrial respiration. Irreversible cell injury was assessed with lactate dehydrogenase (LDH) release. Virus-infected myocytes expressed significantly more vinculin in the cytoskeletal fraction and increased the expression of paxillin but sustained the same amount of injury in response to simulated I/R as control cells (n = 4; P = not significant, paired t-test). Hypothermic I/R (ischemia at 25 degrees C) resulted in a significant reduction in LDH release (P </= 0.02; n = 4). Virus-mediated overexpression of vinculin does not appear to represent a rational approach to overcoming I/R injury.