红景天苷对缺氧缺糖诱导的星形胶质细胞线粒体自噬的影响

摘要

目的:探讨红景天苷(Sal)对缺氧缺糖(OGD)诱导的星形胶质细胞线粒体自噬的影响.方法:提取原代培养的新生鼠皮层星形胶质细胞,进行OGD干预,建立OGD细胞模型.将实验分为对照组、OGD组和Sal+OGD组.用噻唑蓝(MTT)法检测细胞存活率,乳酸脱氢酶(LDH)法检测LDH漏出率评估细胞损伤,Western blot法检测自噬相关蛋白表达水平.结果:Sal终浓度为75、150、300μmol/Ml时,细胞存活率分别为83.15%±5.42%、86.75%±4.91%、91.41%±5.38%,明显高于OGD组(71.22%±8.13%),但仍低于对照组(99.39%±2.17%),差异均有统计学意义(P<0.05);LDH漏出率为34.61%±5.19%、30.51%±8.15%、27.34±7.41%,明显低于OGD组(40.51%±8.15%),但仍高于对照组(16.68%±3.69%),差异均有统计学意义(P<0.05);Western blot结果显示Sal+OGD组中LC3、Beclin1、PINK1和Parkin蛋白表达较OGD组明显降低,但仍高于对照组(P<0.05).结论:Sal抑制OGD诱导线粒体自噬相关蛋白表达,降低OGD诱导的星形胶质细胞损伤.%Objective: To investigate the effect of salidroside (Sal) on mitophagy induced by oxygen-glucose deprivation (OGD) in astrocytes. Methods: The primary astrocytes were isolated from cerebral cortex of neonatal rats and subjected to OGD for imitating hypoxia and glucose deprivation model in vitro. Cells were divided into sham group, OGD model group and Sal pretreatment+OGD group (Sal+OGD). Cell survival rate was detected with MTT and LDH leakage was analyzed with lactate dehydrogenase activity assay kit. Western blot was used to detect the expression of autophagy related protein. Results: The survival rate of cells with final concentra-tion of 75, 150, 300 mol/mL Sal pretreatment and OGD were 83.15%±5.42%, 86.75%±4.91%, 91.41%±5.38%, which were significantly higher than that of the model group (71.22%±8.13%) (P<0.05), but still lower than that of the control group (99.39%±2.17%), the difference was statistically significant (P<0.05). Meanwhile, the LDH leakage rate of cell with final concentration of 75, 150, 300 mol/mL Sal pretreatment were 34.61%±5.19%, 30.51%±8.15%, 27.34%±7.41%, which was significantly lower than that of model group (40.51%±8.15%), but still higher than that of the control group (16.68%±3.69%), the difference was statistically significant (P<0.05). Western blot showed the expression of LC3, Beclin1, PINK1 and Parkin protein in the Sal+OGD group significantly lower than that in model group, but still higher than that in the control group (P<0.05). Conclusion:Sal decreases mitophagy related protein and restored the cell proliferation inhibition induced by OGD in astrocytes.