姜黄素通过清除小胶质细胞内活性氧类物质保护多巴胺能细胞

Curcumin protects doparminergic cells by inhibitting microglia activation

目的探讨小胶质细胞在多巴胺能细胞损伤中的作用,以及姜黄素通过抑制小胶质细胞反应保护多巴胺能细胞的机制。方法选用大鼠嗜铬细胞瘤株PC12细胞,利用鱼藤酮诱导其损伤建立帕金森病细胞模型,用姜黄素预处理4 h的小胶质细胞(BV-2)再经鱼藤酮处理4 h后的细胞上清作为条件培养液(microglia conditioned medium with curcumin,MCMC)处理PC12细胞,并设空白对照组。应用四甲基偶氮唑盐法(MTT法)检测细胞活力;DCFH-DA染色检测BV-2细胞内活性氧类物质(ROS)水平;磷脂结合蛋白(annexinⅤ)-碘化丙啶(PI)双染色流式细胞仪检测PC12细胞凋亡。结果 5nM鱼藤酮单独作用于PC12细胞,其存活率及凋亡率与空白组相比无明显差别(P>0.05);与对照组相比,姜黄素预处理的BV-2细胞,其ROS水平降低(P<0.01);受MCMC处理的PC12细胞,与对照组比较,细胞活力增加,细胞凋亡率降低(P<0.01)。结论鱼藤酮通过激活小胶质细胞产生ROS,损伤多巴胺能细胞。姜黄素通过抑制小胶质细胞反应,清除小胶质细胞内ROS,从而保护多巴胺能细胞。

Objective To explore the effects of microglia on the impairment of dopaminergic cells and the mecha- nisms of Curcumin＇ s protective effects on them by inhibitting microglia activation. Methods The cellular model of PD was established by adoption of rat pheoehromoeytoma strain PC12 cells treated with Rotenone, PC12 cells were treated with the medium from Curcumin-pretreated-BV-2 cells which were conditioned with Rotenone, and the blank control group was estab- lished. Cell viability was assessed with MTT;DCFH-DA staining was used to measure the level of iutracellular ROS in BV- 2 cells;flow eytometrie analysis to survey the rate of cell apoptosis. Results Compared with the control group, the level of ROS in Curcumin-pretreated-BV-2 cells groups was decreased （P 〈0.01 ） ;Cell viability of PC12 cells treated with MCMC was increased and the rate of apoptosis of them was decreased（ P 〈 0.01 ）. The cell viability and rate of apoptosis of PC12 cells treated only with 5 nM Rotenone were not different with those of blank control group （ P 〉 O. 05 ）. Conclusion Mi- croglia was activated by Rotenone impairing dopaminergic cells by producing ROS. Curcumin protects dopaminergic ceils by decreasing the production of ROS in microglia.