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近来发现新的蛋白激酶Cδ亚型(PKCδ)的磷酸化激活与帕金森病(PD)中神经元的缺失有关.我们的前期研究发现,PKCδ磷酸化参与6-羟基多巴胺引起的多巴胺能细胞的毒性作用,但其具体机制尚不清楚.本研究以TUNEL检测发现,6-羟基多巴胺引起较显著的细胞凋亡,PKCδ抑制剂Rottlerin可减轻6-羟基多巴胺诱导的凋亡,总PKC抑制剂Bis、钙依赖性PKC(α和β)抑制剂G6976对6-羟基多巴胺诱导的凋亡无影响.采用Western免疫印迹杂交实验发现,6-羟基多巴胺持续性激活ERK 1/2和PKCδ,Rottlerin既可抑制PKCδ的磷酸化激活,又可抑制ERK的磷酸化激活,而MEK抑制剂U0126仅能抑制ERK 1/2磷酸化激活,对PKCδ磷酸化却无显著影响.这说明PKCδ是6-羟基多巴胺持续性激活ERK 1/2的上游激酶.本研究结果提示,在凋亡过程中PKCδ仍然是ERK1/2激活的上游激酶,阻断PKCδ磷酸化可阻断ERK1/2持续激活.Rottlerin正是由于阻断PKCδ的激活,进一步阻断ERK1/2持续激活,减轻多巴胺能细胞的凋亡.因此,Rottlerin可能对防治帕金森病患者神经元缺失有一定作用.
It has recently been discovered that the phosphorylation of the novel protein kinase Cδ subtype (PKCδ) is associated with a loss of neurons in Parkinson’s disease (PD). Our previous study found that PKCδ phosphorylation is involved in the 6-hydroxydopamine-induced dopaminergic cells Toxicity, but its specific mechanism is not clear.In this study, TUNEL assay found that 6-hydroxydopamine caused more significant apoptosis, PKCδ inhibitor Rottlerin can reduce 6-hydroxydopamine-induced apoptosis, total PKC inhibitor Bis , And calcium-dependent PKC (α and β) inhibitor G6976 had no effect on 6-hydroxydopamine-induced apoptosis.Western blot analysis showed that 6-hydroxydopamine sustained ERK 1/2 and PKCδ, Rottlerin Which not only inhibited the phosphorylation of PKCδ, but also inhibited the phosphorylation of ERK, while MEK inhibitor U0126 only inhibited the phosphorylation of ERK 1/2 and had no significant effect on the phosphorylation of PKCδ, indicating that PKCδ was 6-hydroxy Dopamine sustained activation of ERK 1/2 upstream kinase.The results suggest that in the process of apoptosis, PKCδ is still an upstream kinase activated by ERK1 / 2, blocking PKCδphosphorylation can block ERK1 / 2 sustained activation.Rottlerin is Due to blocking PKCδ Live, further blocking ERK1 / 2 activation duration, reduce apoptotic dopaminergic cells Thus, Rottlerin, an may have a role in the prevention of neuronal loss with Parkinson’s disease.