Long-term inhibition of RhoA attenuates vascular contractility by enhancing endothelial NO production in an intact rabbit mesenteric artery
Long-term inhibition of RhoA attenuates vascular contractility by enhancing endothelial NO production in an intact rabbit mesenteric artery
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DOI：
10.1161/01.res.0000165483.34603.91
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发表时间：
2005-05-13
影响因子：
20.1
通讯作者：
Kanaide, H
中科院分区：
文献类型：
--
作者：
Shiga, N;Hirano, K;Kanaide, H
RhoA在调节培养的内皮细胞中的NO产生中起关键作用。为了确定其在原位内皮细胞中的作用，我们研究了3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂和Rho激酶（RB）的RhoA结合结构域对离体兔肠系膜动脉血管收缩力的影响。用3 × 10（-5）mol/ L辛伐他汀和氟伐他汀离体处理条约24至30小时，显著减弱了内皮存在时对苯肾上腺素和高K+的收缩反应。加入N-ω-硝基-L-精氨酸甲酯和去除内皮可消除收缩反应的衰减。与香叶基香叶基焦磷酸盐共同处理阻止了他汀引起的收缩反应的衰减，而香叶基香叶基转移酶抑制剂模拟了辛伐他汀的作用。辛伐他汀治疗增强了缓激肽诱导的肠系膜动脉内皮依赖性舒张，而对缓激肽诱导的主动脉瓣内皮细胞[Ca 2 +]（i）升高没有影响。用达特蛋白的细胞穿透肽（TATHA-RB）将RB引入条中以NO依赖的方式减弱收缩反应。然而，Rac 1/Cdc 42结合片段的p21激活的蛋白激酶，RB没有达特肽或TATHA蛋白A没有影响。辛伐他汀和TATHA-RB在体内处理兔以NO依赖的方式减弱收缩力。辛伐他汀和TATHA-RB显着上调eNOS在兔肠系膜动脉。本研究提供了第一个证据表明，RhoA在抑制原位内皮细胞NO产生中起着生理作用。
RhoA plays a critical role in regulating NO production in cultured endothelial cells. To determine its role in in situ endothelial cells, we investigated the effects of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase inhibitors and a RhoA-binding domain of Rho-kinase (RB) on vascular contractility in the isolated rabbit mesenteric artery. Ex vivo treatment of the strips with 3 x 10(-5) mol/ L simvastatin and fluvastatin for approximate to 24 to 30 hours significantly attenuated the contractile response to phenylephrine and high K+ in the presence of endothelium. The addition of N-omega-nitro-L-arginine methyl ester and the removal of endothelium abolished the attenuation of the contractile response. The cotreatment with geranylgeranyl pyrophosphate prevented thestatin-induced attenuation of the contractile response, whereas geranylgeranyl transferase inhibitor mimicked the effect of simvastatin. Treatment with simvastatin enhanced the bradykinin-induced endothelium-dependent relaxation in the mesenteric artery, whereas it had no effect on the bradykinin-induced [Ca2+](i) elevation in endothelial cells of the aortic valves. Introduction of RB to the strips using a cell-penetrating peptide of Tat protein (TATHA-RB) attenuated the contractile responses in a NO-dependent manner. However, a Rac1/Cdc42-binding fragment of p21-activated protein kinase, RB without Tat peptide or TATHA-protein A had no effect. The in vivo treatment of rabbit with simvastatin and TATHA-RB attenuated the contractility in a NO-dependent manner. Simvastatin and TATHA-RB significantly upregulated eNOS in the rabbit mesenteric artery. The present study provides the first evidence that RhoA plays a physiological role in suppressing NO production in in situ endothelial cells.