磷酸化PKCδ参与6-OHDA诱导的多巴胺能神经细胞凋亡

目的： 观察磷酸化PKCδ在6-OHDA诱导的大鼠PC12细胞株凋亡过程中所起的作用，探讨帕金森病发病的可能分子机制。方法： 体外培养多巴胺能神经细胞株PC12细胞，观察PKC激动剂和抑制剂对6-OHDA毒性作用的影响，分别用TUNEL染色和透射电镜法观察细胞凋亡，Western免疫印迹法观察磷酸化PKCδ的表达。结果： PKCδ激活剂PMA加重6-OHDA的毒性作用，使细胞凋亡率上升。PKCδ抑制剂Rottlerin可抑制PKCδ的磷酸化，减轻6-OHDA引起的细胞凋亡。总PKC抑制剂Bis和钙依赖性PKC抑制剂G6976不能减轻6-OHDA引起的细胞凋亡。结论： 在6-OHDA诱导PC12细胞凋亡的过程中，PKCδ的磷酸化是重要环节之一，PKCδ可能直接参与帕金森病人的神经元凋亡过程。

Phosphorylation PKCδ participates in apoptosis of PC12 cells induced by 6-hydroxydopamine

AIM: To observe the effect of phosphorylation protein kinase C delta (PKCδ) on the procedure of PC12 cells apoptosis induced by 6-hydroxydopamine(6-OHDA) and to investigate the potential molecular pathogenesis of Parkinson disease.METHODS: TUNEL staining and transmission electron microscope were applied to measure apoptosis when dopaminergic PC12 cells exposed to the excitomotors and inhibitors of PKC before 6-OHDA for 18 hours. The expression of phosphorylation of PKCδ was detected by Western blotting. RESULTS: PMA, an activating agent of PKCδ, significantly increased PC12 cell apoptosis induced by 6-OHDA. Rottlerin, an inhibitor of PKCδ, protected PC12 cells apparently. As contrast, bisindolylmaleimide I, an inhibitor of general PKC and G6976, the inhibitor of calcium-dependent PKC, did not show any protective role. CONCLUSION: The phosphorylation PKCδ is one of the important links in the process of PC12 cell apoptosis induced by 6-OHDA. PKCδ may directly participate in neurodegeneration process in parkinsonian.