黄芪甲苷抑制缺氧缺糖复氧复糖PC12细胞凋亡的研究

Study on Astragaloside Ⅳ Inhibiting PC12 Apoptosis Induced by Oxygen-glucose Deprivation and Restoration

目的:研究黄芪甲苷对缺氧缺糖复氧复糖PC12细胞凋亡的抑制作用。方法:取对数期PC12细胞随机分为6组:正常对照组(Normal组)、黄芪甲苷溶剂对照组(DMSO组)、模型组(缺氧缺糖复氧复糖组,Model组)、黄芪甲苷高剂量组(AST H组,100μmol/L)、黄芪甲苷中剂量组(AST M组,50μmol/L)、黄芪甲苷低剂量组(AST L组,25μmol/L)。正常对照组正常培养不进行任何处理;其余各组均进行缺氧缺糖复氧复糖造模:缺氧缺糖4 h后复氧复糖24 h;黄芪甲苷各剂量组和溶剂对照组于造模前0.5 h给药,直至培养结束。用MTT方法检测细胞存活率,倒置显微镜下观察细胞的形态,流式细胞仪检测细胞凋亡情况。结果:与正常组相比,模型组细胞折光性差,细胞肿胀簇状聚集,甚至脱落,细胞之间的突触消失,细胞本身明显肿胀,存活率明显降低,细胞凋亡率增加(P<0.05)。与模型组相比,溶剂对照组和黄芪甲苷低剂量组细胞形态无明显变化,存活率和凋亡率差异没有显著性(P>0.05),但黄芪甲苷高、中剂量组细胞肿胀等变化明显减轻,存活率均增加,细胞凋亡率降低(P<0.05)。黄芪甲苷高剂量组比中、低剂量组细胞存活率高和凋亡率低(P<0.05)。结论:黄芪甲苷可抑制PC12细胞缺氧缺糖复氧复糖后的损伤和凋亡,其最佳作用浓度是100μmol/L。

Objective: to study the inhibition of Astragaloside Ⅳ on PC12 apoptosis induced by oxygen-glucose deprivation and restoration. Methods: PC12 cells of logarithmic phase were divided randomly into six groups: Normal group,DMSO group( Astragaloside IV solvent),Model group( Oxygen-glucose Deprivation and Restoration),ASTH group( high dose of Astragaloside IV,100μmol /L),ASTM group( medium dose,50 μmol /L),and ASTL group( low dose,25 μmol /L). Normal group were cultivated normally without any processing,while the others were making models with oxygen-glucose deprivation and restoration: 4 hours' deprivation before restoring oxygen and glucose for 24 hours; administrations to all dose groups and DMSO group started 0. 5 hour before model making and it didn' t stop till the end of cultivation. Cell survival rate was detected with MIT,cell morphology observed under the inverted microscope,and cell apoptosis detected with flow cytometer. Results: compared with normal group,cells of model group had inferior refraction,swelling,clustering even ablating; synapses between cells disappeared,the survival rate significantly decreased and the apoptotic rate increased( P < 0. 05). Compared with model group,the morphology had no obvious changes in DMSO group and all dose groups,and there was no significant difference in survival rate and apoptotic rate( P > 0. 05),but in ASTH and ASTM groups,cells were less swelling,the survival rate decreased and the apoptotic rate decreased( P < 0. 05). Of ASTH group,the survival rate was higher and the apoptotic rate was lower than those of ASTM and ASTL group( P < 0. 05). Conclusion: Astragaloside IV can inhibit the injury and apoptosis of PC12 induced by oxygen-glucose deprivation and restoration,and the best activity is 100μmol /L.