首页> 外文期刊>American Journal of Physiology >Mechanism of RhoA/Rho kinase activation in endothelin-1- induced contraction in rabbit basilar artery.
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Mechanism of RhoA/Rho kinase activation in endothelin-1- induced contraction in rabbit basilar artery.

机译:RhoA / Rho激酶活化在内皮素-1诱导的兔基底动脉收缩中的机制。

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摘要

This study was undertaken to demonstrate the role of the RhoA/Rho kinase pathway in endothelin-1 (ET-1)-induced contraction of the rabbit basilar artery. Isometric tension and Western blot were used to examine ET-1-induced contraction and RhoA activation. The upstream effect on ET-1-induced RhoA activity was determined by using ET(A) and ET(B) receptor antagonists, protein kinase C (PKC), tyrosine kinase, and phosphatidylinositol-3 kinase inhibitors. The downstream effect of ET-1-induced contraction and RhoA activity was studied in the presence of the Rho kinase inhibitor Y-27632. The effect of Rho kinase inhibitor on ET-1-induced myosin light chain (MLC) phosphorylation was investigated by using urea-glycerol-PAGE immunoblotting. We found 1) ET-1 increased RhoA activity (membrane binding RhoA) in a concentration-dependent manner; 2) ET(A), but not ET(B), receptor antagonist abolished the effect of ET-1 on RhoA activation; 3) phosphodylinositol-3 kinase inhibitor, but not PKC and tyrosine kinase inhibitors, reduced ET-1-induced RhoA activation; 4) Rho kinase inhibitor Y-27632 (10 microM) inhibited ET-1-induced contraction; and 5) ET-1 increased the level of MLC phosphorylation. Rho kinase inhibitor Y-27632 reduced the effect of ET-1 on MLC phosphorylation. This study demonstrated that RhoA/Rho kinase activation is involved in ET-1-induced contraction in the rabbit basilar artery. Phosphodylinositol-3 kinase and MLC might be the upstream and downstream factors of RhoA activation.
机译:进行这项研究以证明RhoA / Rho激酶途径在内皮素1(ET-1)诱导的兔基底动脉收缩中的作用。使用等轴测张力和蛋白质印迹法检查ET-1诱导的收缩和RhoA激活。通过使用ET(A)和ET(B)受体拮抗剂,蛋白激酶C(PKC),酪氨酸激酶和磷脂酰肌醇3激酶抑制剂确定对ET-1诱导的RhoA活性的上游影响。在Rho激酶抑制剂Y-27632存在下研究了ET-1诱导的收缩和RhoA活性的下游作用。通过使用尿素-甘油-PAGE免疫印迹研究了Rho激酶抑制剂对ET-1诱导的肌球蛋白轻链(MLC)磷酸化的影响。我们发现1)ET-1以浓度依赖性方式增加RhoA活性(膜结合RhoA); 2)ET(A)而不是ET(B)受体拮抗剂消除了ET-1对RhoA激活的影响; 3)磷酸化肌醇3激酶抑制剂,而不是PKC和酪氨酸激酶抑制剂,降低了ET-1诱导的RhoA活化; 4)Rho激酶抑制剂Y-27632(10 microM)抑制ET-1诱导的收缩; 5)ET-1增加了MLC磷酸化水平。 Rho激酶抑制剂Y-27632降低了ET-1对MLC磷酸化的作用。这项研究表明RhoA / Rho激酶激活与ET-1诱导的兔基底动脉收缩有关。磷酸肌醇-3激酶和MLC可能是RhoA激活的上游和下游因素。

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