摘要
目的 为了探讨钙离子通道阻断剂维拉帕米是否会对母鸡口服三邻甲苯基磷酸酯 (TOCP)后脑干微粒体蛋白磷酸化有影响。方法 维拉帕米的给药剂量为 7mg·kg- 1(im) ,给药共 4d(d 1～d 4 ) ,d2igTOCP 75 0mg·kg- 1。体外实验测定 [γ 32 P]ATP磷酸基团对蛋白质的渗入量来表示蛋白质磷酸化的强弱 ,利用十二烷基硫酸钠聚丙烯酰胺凝胶电泳方法分离放射性标记的蛋白质并用放射自显影术来显示磷酸化的蛋白质 ,蛋白质磷酸化的变化以吸光度值的变化来定量。结果 给予TOCP后 ,鸡脑干微粒体蛋白的整体磷酸化水平增强 ,特别是 4 5、4 1和32ku蛋白质的磷酸化水平分别增加至对照组的118.7%、173.7%、172 .7% ;而维拉帕米则能消除TOCP处理组磷酸化水平的增加。结论 维持脑干微粒体蛋白磷酸化水平的平衡可能是钙通道阻断剂缓解迟发性神经毒性的机理之一。
AIM To examine if the calcium channel blocker, verapamil, would affect the level of brainstem microsomal protein phosphorylation after administration of tri o cresyl phosphate(TOCP), by which maybe indicate some clues to the pathogenesis of organophosphate induced delayed polyneuropathy(OPIDP). METHODS Verapamil(7 mg·kg -1 ·d -1 , im) was given for 4 d (d 1-d 4) . A dose of TOCP(750 mg·kg -1 ) was administrated by ig on d 2. Phosphorylation of brainstem microsomal proteins was assayed in vitro by using [γ 32 P]ATP as phosphate donor. Radiolabeled proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and visualized by autoradiography. The changes of the protein phosphorylation were quantified by densitometry. RESULTS TOCP administration enhanced the phosphorylation of the microsomal proteins with apparent molecular weights of 45, 41, 32 ku by 119%, 174%, 173%, respectively. Verapamil abolished the enhancement induced by TOCP. CONCLUSION The mechanism of calcium channel blocker to ameliorate organophosphate induced delayed polyneuropathy might be the protection of the brainstem microsomal proteins from enhancement of phosphorylation.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2002年第3期165-171,共7页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金项目 (39770 15 1)
国家自然科学基金项目 (39970 12 7)
中国科学院生物科学与生物技术研究特别支持费课题 (财政部专项
STZ98213)~~
关键词
维拉帕米
三邻甲苯基磷酸酯
脑干
微粒体蛋白
磷酸化
organophosphorus compound, tri o cresyl phosphate
verapamil
microsomes
phosphorylation
