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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >Acetylcholine-induced phosphorylation of CPI-17 in rat bronchial smooth muscle: the roles of Rho-kinase and protein kinase C.
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Acetylcholine-induced phosphorylation of CPI-17 in rat bronchial smooth muscle: the roles of Rho-kinase and protein kinase C.

机译:乙酰胆碱诱导的大鼠支气管平滑肌CPI-17磷酸化:Rho激酶和蛋白激酶C的作用。

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摘要

It has been demonstrated that CPI-17 provokes an inhibition of myosin light chain phosphatase to increase myosin light chain phosphorylaton and Ca(2+) sensitivity during contraction of vascular smooth muscle. However, expression and agonist-mediated regulation of CPI-17 in bronchial smooth muscle have not been documented. Thus, expression and phosphorylation of CPI-17 mediated by PKC and ROCK were investigated using rat bronchial preparations. Acetylcholine (ACh)-induced contraction and Ca(2+) sensitization were both attenuated by 10(-6) mol Y-27632 /L, a ROCK inhibitor, 10(-6) mol calphostin C/L, a PKC inhibitor, and their combination. A PKC activator, PDBu, induced a Ca(2+) sensitization in alpha-toxin-permeabilized bronchial smooth muscle. In this case, the Ca(2+) sensitizing effect was significantly inhibited by caphostin C but not by Y-27632. An immunoblot study demonstrated CPI-17 expression in the rat bronchial smooth muscle. Acetylcholine induced a phosphorylation of CPI-17 in a concentration-dependent manner, which was significantly inhibited by Y-27632 and calphostin C. In conclusion, these data suggest that both PKC and ROCK are involved in force development, Ca(2+) sensitization, and CPI-17 phosphorylation induced by ACh stimulation in rat bronchial smooth muscle. As such, RhoA/ROCK, PKC/CPI-17, and RhoA/ROCK/CPI pathways may play important roles in the ACh-induced Ca(2+) sensitization of bronchial smooth muscle contraction.
机译:已经证明,CPI-17会抑制肌球蛋白轻链磷酸酶,从而增加血管平滑肌收缩过程中的肌球蛋白轻链磷酸化和Ca(2+)敏感性。但是,尚未证明支气管平滑肌中CPI-17的表达和激动剂介导的调节。因此,使用大鼠支气管制剂研究了由PKC和ROCK介导的CPI-17的表达和磷酸化。乙酰胆碱(ACh)诱导的收缩和Ca(2+)致敏性都被10(-6)mol Y-27632 / L,ROCK抑制剂,10(-6)mol calphostin C / L,PKC抑制剂和他们的组合。一个PKC激活剂,PDBu,在α-毒素透化的支气管平滑肌中诱导Ca(2+)致敏。在这种情况下,Ca(2+)致敏作用被caphostin C显着抑制,但不受Y-27632抑制。一项免疫印迹研究表明,CPI-17在大鼠支气管平滑肌中表达。乙酰胆碱以浓度依赖性的方式诱导CPI-17的磷酸化,该磷酸化被Y-27632和钙磷蛋白C显着抑制。总之,这些数据表明PKC和ROCK均参与力发展,Ca(2+)致敏,以及ACh刺激在大鼠支气管平滑肌中诱导的CPI-17磷酸化。因此,RhoA / ROCK,PKC / CPI-17和RhoA / ROCK / CPI途径可能在ACh诱导支气管平滑肌收缩的Ca(2+)致敏中起重要作用。

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