龙血通络胶囊对H2O2诱导PC12细胞损伤的保护作用

Protective Effect of Longxue Tongluo Capsule on H2O2-induced Injury of PC12 Cells

目的:本文探讨过氧化氢(H2O2)诱导PC12细胞损伤后,龙血通络胶囊的神经保护作用及其潜在作用机制。方法:以500μmol·L-1H2O2构建PC12细胞损伤模型,实验分为空白组,模型组(H2O2,500μmol·L^-1),龙血通络胶囊组(LTC,1,2,4 mg·L^-1)。通过细胞增殖毒性检测(CCK-8),胞内活性氧(ROS)水平,线粒体膜电位水平(JC-1),细胞凋亡率以及蛋白免疫印迹法(Western blot)评价龙血通络胶囊对H2O2诱导PC12细胞损伤的神经保护作用。结果:与空白组比较,模型组的细胞存活率显著降低(P<0.01),ROS水平显著上升(P<0.01),线粒体膜电位水平下降,细胞凋亡率显著升高(P<0.01),剪切的聚腺苷二磷酸核糖聚合酶(cleaved PARP)水平显著上升(P<0.01)。与模型组比较,LTC组(1,2,4 mg·L^-1)能浓度依赖性升高细胞的存活率(P<0.01),降低ROS水平(P<0.05),维持线粒体膜电位水平,降低细胞的凋亡水平(P<0.01),并显著降低PARP蛋白的剪切水平(P<0.01)。结论:龙血通络胶囊对H2O2诱导的PC12细胞损伤具有显著的保护作用,其作用机制可能是通过抑制胞内氧化应激,维持线粒体功能,促进DNA修复等途径进而抑制神经细胞凋亡。

Objective:To investigate the protective effect of Longxue Tongluo capsule(LTC)on H2O2-induced injury of PC12 cells and its potential mechanism.Method:An in vitro injury model in PC12 cells was established with 500μmol·L-1H2O2.The experiment was divided into control group,injury model group(H2O2500μmol·L-1),and Longxuetongluo capsule group(LTC,1,2,4 mg·L-1).Cell counting kit-8(CCK-8)assay,intracellular reactive oxygen species(ROS),mitochondrial membrane potential,apoptosis of PC12 cells,and Western blot were used to evaluate the protective effect of LTC on PC12 cells induced by H2O2.Result:Compared with the control group,the cell viability was significantly decreased in the injury model group(P<0.01),intracellular ROS level was significantly increased(P<0.01),mitochondrial membrane potential was decreased,while apoptosis of PC12 cells was significantly increased(P<0.01),and the expression of cleaved poly adenosine diphospho ribose polymerase(PARP)was also increased significantly(P<0.01).Compared with the injury model group,pretreatment with LTC at the concentrations of 2 and 4 mg·L for 6 h significantly increased cell viability in PC12 cells exposed to H2O2(P<0.01).Moreover,pretreatment with LTC reduced intracellular ROS level(P<0.05),maintained mitochondrial membrane potential,and inhibited apoptosis of PC12 cells induced by H2O2in a dose-dependent manner(P<0.01).The results of western blotting showed that pretreatment with LTC significantly reduced the expression of cleaved PARP(P<0.01).Conclusion:LTC exerts a significant protective effect against H2O2-induced PC12 cells injury through inhibition of neuronal apoptosis by suppressing intracellular oxidative stress,maintaining mitochondrial function,and promoting DNA repair.