基础医学与临床 ›› 2007, Vol. 27 ›› Issue (6): 610-615.

• 研究论文 • 上一篇    下一篇

缺血预处理对大鼠视网膜内cPKC亚型特异性膜转位和蛋白表达量的影响

丁宁 王宁利 魏勇 李俊发   

  1. 首都医科大学附属北京同仁医院 首都医科大学北京同仁眼科中心 首都医科大学附属北京同仁医院 首都医科大学神经生物系
  • 收稿日期:2007-02-05 修回日期:2007-03-09 出版日期:2007-06-25 发布日期:2007-06-25
  • 通讯作者: 丁宁

Effect of ischemic pretreatment on levels of cPKC isoform-specific membrane translocation and protein expression in retina of rats

  

  • Received:2007-02-05 Revised:2007-03-09 Online:2007-06-25 Published:2007-06-25

摘要: 目的 通过观察缺血预处理(IP)对大鼠视网膜内经典型蛋白激酶C(cPKC)a、bI、bII和g等4种亚型特异性膜转位(激活)水平和蛋白表达量的影响,确定可能参与视网膜缺血预适应(IPC)形成的cPKC亚型。方法 体重200~250g 雄性Wistar大鼠随机分为正常对照组(C,n=6),IP组(左眼内压17.29kPa并维持5min,n=48),假手术组(Sham,n=48);后两组分别在手术处理后10、20和40min及1、12、24、72和168h(每个时间点n=6)取视网膜组织。应用Western blot检测cPKC亚型特异性膜转位水平和蛋白表达量。结果 大鼠视网膜组织内cPKCa、bI、bII和g等亚型中,cPKCa膜转位水平在IP处理后10min~168h内与C和Sham组相比无显著改变,而蛋白表达量在IP处理后12~168h明显升高,在72h达高峰。cPKCg膜转位水平在IP处理后20min~1h内显著增高,在40min达高峰;cPKCg蛋白表达量在IP处理后12~72h明显增高,在24h达高峰。IP处理对视网膜组织内cPKCbI、bII膜转位水平和蛋白表达量均无显著影响。结论 cPKCg膜转位(激活)增强可能参与大鼠视网膜IPC早期形成,而cPKCa和g蛋白表达量的增高可能与晚期IPC的维持有关。

Abstract: Objective To identify certain isoforms involved in the onset of retinal ischemic preconditioning (IPC), the effects of ischemic pretreatment (IP) were observed on levels of conventional protein kinase C (cPKC) a, bI, bII and g isoform-specific membrane translocation and protein expression in retina of rats. Methods Retinal IP was produced by intra-ocular pressure (IOP) elevation for 5 minutes in anesthetized Wistar rats. Sham operation was similar to IP except the pressure elevation. 10, 20, or 40 minutes and 1, 12, 24, 72, or 168 hours after the procedure, levels of cPKC isoform-specific membrane translocation and protein expression were analyzed using Western-blot. Results cPKCa protein expression levels increased significantly from 12h to 168h. A peak reached at 72h after IP. cPKCg membrane translocation enhanced during 20min to 1h with a peak at 40min. cPKCg protein expression levels increased significantly from 12h to 72h. A peak was at 24h after IP. However, there were no significant changes in both membrane translocation of cPKCa, bI, bII and protein expression of cPKCbI, bII in retina of rats following IP. Conclusion The enhanced cPKCg membrane translocation, and the increased cPKCa and g protein expressions might be involved in the onset and sustain of retinal IPC in rats respectively.