Cytoskeletal activation and altered gene expression in endothelial barrier regulation by simvastatin
Cytoskeletal activation and altered gene expression in endothelial barrier regulation by simvastatin
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DOI：
10.1165/rcmb.2003-0267oc
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发表时间：
2004-05-01
影响因子：
6.4
通讯作者：
Garcia, JGN
中科院分区：
文献类型：
--
作者：
Jacobson, JR;Dudek, SM;Garcia, JGN
他汀类药物是一类HMG-CoA还原酶抑制剂，通过抑制香叶基香叶基化（一种Rho GTdR与细胞膜结合激活剂相互作用所必需的共价修饰）直接影响多种血管过程。我们探讨了辛伐他汀对内皮细胞肌动球蛋白收缩、间隙形成和由水肿剂凝血酶产生的屏障功能障碍的影响。人肺动脉内皮细胞暴露于延长辛伐他汀治疗（5 μ M，16小时）表现出显着减少凝血酶诱导（1 U/ml）的屏障功能障碍（类似于70%的抑制）与加速屏障恢复，作为测量的跨内皮阻力。此外，辛伐他汀衰减基础和凝血酶刺激（1 U/ml，5分钟）肌球蛋白轻链二磷酸化和应力纤维的形成，同时显着增加外周免疫染色的肌动蛋白和corpine，肌动蛋白结合蛋白，结合增加Rac GTdR活性。由于辛伐他汀诱导的Rac激活和屏障保护延迟（最大16小时后），我们评估了基因表达和蛋白质翻译在辛伐他汀反应中的作用。同时用放线菌酮（10 μ g/ml，16小时）治疗取消辛伐他汀介导的屏障保护。通过微阵列分析，观察到细胞骨架蛋白（钙调蛋白、整联蛋白[14]）、凝血酶调节元件（帕拉、血栓调节蛋白）和信号传导基因（鸟嘌呤核苷酸交换因子）的表达对辛伐他汀的反应发生了显著变化。这些新的观察结果在许多血管病理学中具有广泛的临床意义，其特征在于血管完整性的改变，包括炎症、血管生成和急性肺损伤。
The statins, a class of HMG-CoA reductase inhibitors, directly affect multiple vascular processes via inhibition of geranylgeranylation, a covalent modification essential for Rho GTPase interaction with cell membrane-bound activators. We explored simvastatin effects on endothelial cell actomyosin contraction, gap formation, and barrier dysfunction produced by the edemagenic agent, thrombin. Human pulmonary artery endothelial cells exposed to prolonged simvastatin treatment (5 muM, 16 h) demonstrated significant reductions in thrombin-induced (1 U/ml) barrier dysfunction (similar to 70% inhibition) with accelerated barrier recovery, as measured by transendothelial resistance. Furthermore, simvastatin attenuated basal and thrombin-stimulated (1 U/ml, 5 min) myosin light chain diphosphorylation and stress fiber formation while dramatically increasing peripheral immunostaining of actin and cortactin, an actin-binding protein, in conjunction with increased Rac GTPase activity. As both simvastatin-induced Rac activation and barrier protection were delayed (maximal after 16 h), we assessed the role of gene expression and protein translation in the simvastatin response. Simultaneous treatment with cycloheximide (10 mug/ml, 16 h) abolished simvastatin-mediated barrier protection. Robust alterations were noted in the expression of cytoskeletal proteins (caldesmon, integrin [14), thrombin regulatory elements (PARA, thrombomodulin), and signaling genes (guanine nucleotide exchange factors) in response to simvastatin by microarray analysis. These novel observations have broad clinical implications in numerous vascular pathobiologies characterized by alterations in vascular integrity including inflammation, angiogenesis, and acute lung injury.