Inhibition of Rho-kinase leads to rapid activation of phosphatidylinositol 3-kinase protein kinase Akt and cardiovascular protection
Inhibition of Rho-kinase leads to rapid activation of phosphatidylinositol 3-kinase protein kinase Akt and cardiovascular protection
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DOI：
10.1161/01.atv.0000142813.33538.82
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发表时间：
2004-10-01
影响因子：
8.7
通讯作者：
Liao, JK
中科院分区：
文献类型：
--
作者：
Wolfrum, S;Dendorfer, A;Liao, JK
目的-Rho-Kinase活性在心血管疾病和有心血管危险因素的患者中升高。方法与结果：在人内皮细胞中，抑制Rho激酶抑制剂羟基法舒地尔(HF)(1至100mumol/L)可在15分钟内增加Akt丝氨酸473的磷酸化，导致Akt激酶活性和一氧化氮(NO)释放分别增加2.2倍和4.0倍。磷脂酰肌醇3-激酶(PI3-K)抑制剂LY294002(10 mumol/L)可完全阻断HF对Akt和eNOS的激活。为了确定这一通路的生理学相关性，我们使用了两种缺血-再灌注(I/R)损伤模型。急性给予法舒地尔(10 mg/kg，腹腔注射，缺血前1小时)可减少野生型小鼠I/R损伤后的白细胞募集和与肠系膜内皮细胞的黏附，但对eNOS(-/-)小鼠无影响。同样，法舒地尔治疗可使冠状动脉短暂闭塞的大鼠的心肌梗死面积减少38%。联合应用2种PI3-激酶抑制剂Wortmannin和LY294002或eNOS抑制剂L-NAME可阻断法沙地尼的心血管保护作用。结论：抑制Rho-Kinase可激活PI3-K/Akt/eNOS通路，从而发挥心血管保护作用。提示Rho-Kinase可能在介导I/R损伤的炎症反应中发挥重要作用。
Objective - Rho-Kinase activity is increased in cardiovascular diseases and in patients with cardiovascular risk factors. However, it is not known whether inhibition of Rho-kinase could lead to cardiovascular protection and, if so, by what mechanism.Methods and Results - In human endothelial cells, the Rho-kinase inhibitor, hydroxyfasudil (HF) ( 1 to 100 mumol/L), increased Akt serine-473 phosphorylation within 15 minutes, leading to a 2.2-fold and 4.0-fold increase in Akt kinase activity and nitric oxide ( NO) release, respectively. Activation of Akt and eNOS by HF was completely blocked by the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, LY294002 ( 10 mumol/L). To determine the physiological relevance of this pathway, we used 2 models of ischemia-reperfusion (I/R) injury. Acute administration of fasudil ( 10 mg/kg, intraperitoneal, 1 hour before ischemia) decreased leukocyte recruitment and adhesion to the mesenteric endothelium after I/R injury in wild-type but not eNOS(-/-) mice. Similarly, treatment with fasudil decreased myocardial infarct size by 38% in rats subjected to transient coronary artery occlusion. Cotreatment with 2 PI3-kinase inhibitors, wortmannin and LY294002, or the eNOS inhibitor, L-NAME, blocked the cardiovascular protective effects of fasudil.Conclusions - Inhibition of Rho-kinase leads to the activation of the PI3-kinase/Akt/eNOS pathway and cardiovascular protection. These findings suggest that Rho-kinase may play an important role in mediating the inflammatory response to I/R injury.