右美托咪定对PC12细胞氧糖剥夺损伤的保护作用

Protective effects of dexmedetomidine on P12 cells following oxygen-glucose deprivation injury

目的:研究右美托咪定对PC12细胞氧糖剥夺(OGD)损伤的保护作用及机制。方法:采用缺氧缺糖诱导PC12细胞损伤建立OGD损伤细胞模型,将OGD损伤的细胞分为右美托咪定低、中、高浓度组,溶剂组及模型组,并将正常培养的细胞作为正常对照组。各组细胞处理后采用MTT法检测细胞存活率,相应试剂盒分别检测乳酸脱氢酶(LDH)释放量和丙二醛(MDA)含量,AnnexinⅤ-FITC/PI双染法检测细胞凋亡率,Westernblot检测各组细胞中CleavedCaspase-3和PI3K/AKT信号通路相关蛋白总AKT(t-AKT)、磷酸化的AKT(p-AKT)蛋白表达水平。结果:与正常对照组相比,模型组细胞存活率降低,而上清中LDH释放量和MDA含量增多,细胞凋亡率、CleavedCaspase-3和p-AKT蛋白表达水平升高(P<0.05)。与模型组相比,右美托咪定能够上调细胞存活率,降低LDH释放量和MDA含量,抑制CleavedCaspase-3和p-AKT蛋白表达水平(P<0.05)。结论:右美托咪定可能通过抑制PI3K/AKT信号通路的活化对氧糖剥夺损伤神经细胞发挥保护作用。

Aim :To study the protective effects and mechanism of dexmedetomidine on neuronal oxygen glucose deprivation(OGD) injury. Methods :OGD model was established by inducing PC12 cells injury by hypoxia and hypoglycemia. The cells damaged by OGD were divided into low-, medium- and high- concentration groups of dexmedetomidine,solvent group and model group. The normal cultured cells were used as normal control group. Cell viability was detected by MTT assay. The release of lactate dehydrogenase(LDH) and malondialdehyde(MDA) was detected by the kit. Apoptosis was detected by Annexin Ⅴ-FITC/PI staining. Western blot was used to detect the expression levels of Cleaved Caspase-3, t-AKT and p-AKT in each group. Results :Compared with the normal control group, the cell survival rate of the model group decreased, the LDH release and MDA content increased, and the apoptosis rate, and the expression levels of Cleaved Caspase-3 and p-AKT proteins were significantly increased.( P <0.05). Compared with the model group, dexmedetomidine up-regulated cell survival rate, decreased LDH release and MDA content, and inhibited the expressions of Cleaved Caspase-3 and p-AKT proteins( P <0.05). Conclusion :Dexmedetomidine may protect oxygen glucose deprivation injury neural cells by inhibiting the activation of PI3K/AKT signaling pathway.