芍药苷对6羟基多巴胺诱导的PC12细胞损伤的保护作用研究

Protective effect of Paeoniflorin on 6 hydroxydopamine induced PC12 cell injury

目的探讨芍药苷(PF)对6羟基多巴胺(6-OHDA)损害的PC12细胞的保护作用。方法检测各组6-OHDA诱导PC12损伤细胞相对存活率和乳酸脱氢酶(LDH)活性;采用免疫荧光和免疫印迹技术检测PC12细胞α-突触核蛋白(α-syn)、自噬相关蛋白(LC3-Ⅱ、p62)以及酸敏感离子通道(ASIC1a)的蛋白水平。结果与正常对照组比较,6-OHDA组相对细胞存活率显著下降,LDH活性显著升高(均P<0.01)。与6-OHDA组比较,Pc Tx1组、阿米洛利组、PF各亚组相对细胞存活率均显著升高,LDH活性显著降低(P<0.05~0.01)。与正常对照组比较,6-OHDA组ASIC1a蛋白水平显著升高(P<0.01)。与6-OHDA组比较,Pc Tx1组、阿米洛利组、PF各亚组ASIC1a蛋白水平均显著降低(P<0.05~0.01)。与正常对照组比较,6-OHDA组α-syn、LC3-Ⅱ、p62蛋白水平均显著增加(均P<0.01)。与6-OHDA组比较,Pc Tx1组、阿米洛利组、PF各亚组α-syn、LC3-Ⅱ、p62蛋白水平均显著降低(P<0.05~0.01)。结论 PF可能通过提高α-syn的自噬性降解,对6-OHDA毒素损害的PC12细胞起到保护作用。PF的神经保护作用可能与抑制ASIC1a的活性有关。

Objective To study the protective effects of Paeoniflorin（ PF） on 6 hydroxydopamine（ 6-OHDA）induced PC12 cell injury. Methods The relative cell viability and lactate dehydrogenase（ LDH） activity of each group in 6-OHDA-induced PC12 cells were examined. The levels of α-synuclein, autophagy related protein（ LC3-Ⅱ,p62） and activation of acid-sensing ion channel 1 a（ ASIC1 a） in PC12 cells were detected by western blotting and immunofluorescence method. Results Compared with normal control group,the relative cell viability was significantly lower,and the LDH activity was significantly higher in 6-OHDA group（ all P 〈 0. 01）. Compared with the 6-OHDA group,the relative cell viability in Pc Tx1 group,amiloride group and PF group were significantly higher,and the LDH activity were significantly lower（ P 〈 0. 05-0. 01）. Compared with normal control group,the levels of ASIC1 a in 6-OHDA group was significantly higher（ P 〈 0. 01）. Compared with 6-OHDA group,the level of ASIC1 a in Pc Tx1 group,amiloride group and PF group were significantly lower（ P 〈 0. 05-0. 01）. Compared with normal control group,the levels of α-synuclein,LC3-Ⅱ,p62 in the 6-OHDA group were significantly higher（ all P 〈 0. 01）. Compared with 6-OHDA group,the levels of α-synuclein,LC3-Ⅱ,p62 were significantly lower in Pc Tx1 group,amiloride group and PF group（ P 〈 0. 05-0. 01）. Conclusions PF may enhance the autophagic degradation of α-synuclein,thereby protecting PC12 cells damaged by 6-OHDA toxin. The neuroprotective effect of PF may be related to the inhibition of ASIC1 a activity.