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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。
Objective To investigate whether calcium channel blocker verapamil affects the phosphorylation of brain stem microsomal proteins after oral administration of tricresyl phosphate (TOCP) in hens. Methods verapamil was administered at a dose of 7 mg · kg-1 (im) for 4 days (d 1 ~d 4) and d 2igTOCP 75 0 mg · kg -1. In vitro experiments measured [γ 32 P] ATP phosphate groups infiltrated into the protein to indicate the strength of protein phosphorylation, sodium dodecyl sulfate polyacrylamide gel electrophoresis method for the separation of radiolabeled protein and autoradiography To show the phosphorylated protein, changes in protein phosphorylation are quantified as changes in absorbance values. Results After TOCP was administered, the overall phosphorylation level of microsomal protein of chicken brain stem increased. Especially, the phosphorylation levels of 45, 41 and 32ku protein increased to 118.7%, 173.7% and 172.7% of the control group, respectively. Verapamil abolished the increase of phosphorylation in TOCP-treated group. Conclusions The balance of maintaining the phosphorylation level of microsomal protein in the brainstem may be one of the mechanisms of calcium channel blockers in relieving delayed neurotoxicity.