Role of Rho-kinase in cerebral vasospasm and effect of inhalational anesthetics

为了研究PKC在脑动脉收缩机制中的作用，利用猪大脑动脉，进行了以下实验：1)在SDS-PAGE后，用Western blotting方法研究了11个PKC的异构体。2)已知小的GTP结合蛋白RhoA及其下游效应Rho-Kinase(Rock II)参与了钙增敏。3)用免疫印迹法检测肌球蛋白轻链和磷酸化肌球蛋白轻链。4)用双向电泳法、银染法检测和定量p-肌球蛋白轻链/肌球蛋白轻链比值，揭示了Rho-Rho-K途径对钙增敏的机制。另一种钙增敏机制是由于传统的PKCs、α，βI、βII的存在。

To study the role of PKC in the mechanism of contraction in cerebral artery, using porcine cerebral artery, following experiments were planned :1)The eleven PKC isofomis were explored by Western blotting method following SDS-PAGE.2)It is known that one of small GTP binding protein, RhoA, and its downstream effecor, Rho-kinase (Rock II) participate in the calcium sensitization. The presence of these proteins, distribution were examined by Western blotting.3)Myosin light chain (MLC) and phosphorylated myosin light chain (p-MLC) were subjected to Western Blotting.4)Two dimentional electrophoresis, silver staining were applied to detect and quantify the p-MLC/MLC ratio.The presence of the mechasnism of calcium sensitization through Rho-Rho-kinase pathway was indicated.The other mechanism of calcium sensitization, the PKC -mediated pathway was also indicated due to the presence of the conventional PKCs, α, βI, βII.