丙酮酸乙酯对鱼藤酮诱导的多巴胺能神经细胞损伤的保护作用

摘要

Objective To investigate the protective role of ethyl pyruvate in dopaminergic neuronal damage induced by rotenone.Methods SH-SY5Y cells were treated with rotenone,and ethyl pyruvate with different concentration were pretreated to the cells.The cell viability was determined by MTT method.The apoptotic rate of SH-SY5Y cells were analyzed by flow cytometry after Annexin V/PI staining.The protein expression of LC3 Ⅱ / Ⅰ and P62 were assessed by Western blot.The expression and location of HMGB1 were analyzed by immnofluorescence staining.Results Ethyl pyruvate decreased the apoptotic rate of SH-SY5Y cells induced by rotenone,and increased the cell viability.Ethyl pyruvate inhibited the activation of autophage induced by rotenone,accompanied by the increased expression level of P62.Otherwise,ethyl pyruvate also inhibited cytosolic translocation of HMGB1.Conclusion Ethyl pyruvate could inhibit the activation of autophagy by reducing the cytosolic translocation of HMGB1 and plays a protective role in dopaminergic cell.%目的 建立鱼藤酮(Rotenone,Rot)诱导的SH-SY5Y细胞模型,探讨丙酮酸乙酯(Ethyl pyruvate,EP)对多巴胺能神经细胞的保护作用及机制.方法 Rot作用于SH-SY5Y细胞,构建Rot诱导的PD细胞模型,使用不同浓度的EP作用于PD细胞模型,MTT法检测细胞活力,Annexin V/PI染色检测细胞凋亡,免疫印迹法检测LC3Ⅱ/Ⅰ、P62蛋白的表达,免疫荧光染色观察HMGB1的表达及定位.结果 EP减轻了Rot引起的多巴胺能神经细胞凋亡,并提高了细胞活力;EP减少了Rot诱导的LC3Ⅱ/Ⅰ蛋白表达比例增高,增加P62蛋白的表达,且与EP的水平呈正相关.激光共聚焦显微镜观察到EP抑制了HMGB1的胞浆转位.结论 EP可能通过抑制HMGB1的转位来减少Rot诱导的自噬性损伤,从而对多巴胺能神经细胞起到保护作用.