蛋白激酶C活性在脑血管痉挛过程中变化.docVIP

蛋白激酶C活性在脑血管痉挛过程中变化 　 作者：周任　柏鲁宁　吕健　崔刚　巩守平　师蔚 【摘要】 　　目的 探讨蛋白激酶C(PKC)活性在脑血管痉挛(CVS)过程中的变化及其意义。方法 日本大耳白兔25只，随机分为对照组(5只)与模型组(CVS组，20只)，CVS组又分为1、3、5、8d组，每组5只。采用枕大池2次注血法建立脑血管痉挛动物模型，血管造影观察基底动脉(BA)痉挛，放射免疫法和放射性同位素标记蛋白激酶测定法动态测定血浆内皮素和兔BA管壁PKC活性。结果 枕大池2次注血法可以成功构建脑血管痉挛模型，是一种简便、可靠的方法；模型组血浆中内皮素含量和BA壁内PKC的活性均明显升高，且在5d组达到峰值，与对照组相比差异有统计学意义(Plt;0.05)。结论 PKC活性与CVS的严重程度呈正相关。 【关键词】 脑血管痉挛；蛋白激酶C；内皮素 　　PKC activity change in the process of cerebral vasospasm 　　ABSTRACT: Objective To investigate the change of protein kinase C in the process of cerebral vasospasm (CVS) and evaluate its significance. Methods The subjects were divided into control group (n=5) and CVS group (n=20). CVS group was divided into day 1, day 3, day 5, and day 8 subgroups. Twicehemorrhage method was used to establish the rabbit model, angiography was performed to observe the cerebral vasospasm of the basilar artery, and ET level and PKC activity were measured with radioimmunoassay and radioisotopic protein kinase assay kit. Results The twicehemorrhage rabbit model was a simple and reliable one. ET level in the blood plasma and PKC activity of the basilar artery vessel wall was increased in the CVS group, and reached peak in the day 5 group. The difference was significant (Plt;0.05) compared with that in the control group. Conclusion There is a positive correlation between PKC activity and CVS severity. 　　KEY WORDS: cerebral vasospasm (CVS); protein kinase C (PKC); endothelin (ET) 　　脑血管痉挛(cerebral vasospasm, CVS)是影响蛛网膜下腔出血(subarachnoid hemorrhage, SAH)预后的重要因素[12],其病理生理机制目前尚不十分清楚。近年来研究表明，细胞信号传递系统中的重要信号分子蛋白激酶C(protein kinase C, PKC)可能在SAH后CVS发生发展中起重要作用[3]。PKC是一类Ca2+ 磷脂依赖性蛋白激酶，通过催化多种蛋白质上的Ser/Thr磷酸化，调节细胞的代谢、生长、增殖和分化。在本研究中，我们采用兔枕大池2次注血法成功建立CVS模型,动态观察CVS病理过程中血浆中内皮素(endothelin, ET)和基底动脉(basilar artery, BA)内PKC活性的变化,探讨PKC与CVS的关系,为进一步揭示CVS的病理机制提供新的线索。 　　1 材料与方法 　　1.1 主要试剂 [γ32P]ATP(3000ci/mmol)购自Amersham公司；PKC检测盒购自Promega 公司； 蛋白质定量试剂购自Pierce公司；Triton X100购自Sigma公司；DE52购自Whatman公司；ET放免药盒购自中国人民解放军总医院科技开发中心放免研究所；Cordis 3F