天香丹对人脐静脉内皮细胞氧化损伤的保护作用研究

Protective Effect of Tianxiang Dan on Oxidative Stress of HUVECs

目的:探讨天香丹对tBHP诱导损伤的人脐静脉内皮细胞的保护作用。方法:将HUVECs分为对照组、模型组、阳性对照药(维生素C)组和天香丹剂量组(30、60、90μg/mL)。MTT法筛选tBHP最佳造模浓度,并评价天香丹对内皮细胞氧化损伤的保护作用;微板法和WST法检测各组细胞乳酸脱氢酶(LDH)、还原型谷胱甘肽(GSH)和超氧化物歧化酶(SOD)表达情况;显微镜下观察各组细胞形态学变化,Hoechst 33258荧光染色法检测各组细胞凋亡情况。结果:经tBHP诱导24 h后,模型组细胞存活率、SOD活性和GSH含量较对照组显著降低(P<0.01),LDH释放量和细胞凋亡率明显增加(P<0.01);经维生素C和不同浓度天香丹预处理后,与模型组相比较,各组细胞存活率、SOD活性和GSH含量显著增加,LDH的释放和细胞凋亡率得到减轻(P<0.01)。结论:天香丹具有保护HUVECs免于氧化损伤的作用,其机制可能与抗氧化通路的激活,抗氧化能力的增强及逆转细胞凋亡有关。

Objective:To investigate the protective effect of Tianxiang Dan on human umbilical vein endothelial cells(HUVECs)injured by tert-butyl hydroperoxide(tBHP).Methods:HUVECs were divided into the control group,the model group,the vitamin C group and the Tianxiang Dan groups of low-dose(30μg/ml),medium-dose(60μg/ml)and high-dose(90μg/ml).The optimal concentration of tBHP was screened by MTT,and the protective effect of Tianxiang Dan on oxidative damage of HUVECs was evaluated.Microplate method and WST method were used to detect the expressions of lactate dehydrogenase(LDH),reduced glutathione(GSH)and superoxide dismutase(SOD)in each group.The morphological changes of the cells were observed under a microscope in each group,and the apoptosis was observed by Hoechst 33258 fluorescent staining.Results:After induction by tBHP for 24 hours,the cell survival rate,the activity of SOD and the level of GSH were significantly lower(P<0.01),and the release of LDH and theapoptosis rate were significantly increased in the model group than those in the control group(P<0.01).The cell survival rate,the activity of SOD and the level of GSH were significantly increased,and the release of LDH and the apoptosis rate were reduced after the interventions of vitamin C and after the interventions of different concentrations of Tianxiang Dan compared to those in the model group(P<0.01).Conclusion:Tianxiang Dan has the effect of protecting HUVECs from oxidative stress,and its mechanism may be related to the activation of antioxidant pathways,the enhancement of antioxidant capacity and the reversal of cell apoptosis.