齐墩果酸对损伤血管内皮细胞的保护作用及其机制探讨

Protective effects and mechanisms of oleanolic acid on damaged human umbilical vein endothelial cells

目的:探讨齐墩果酸(oleanic acid,OA)对过氧化氢(hydrogen peroxide,H2O2)诱导人血管内皮细胞(human umbilicalvein endothelial cells,HUVECs)损伤的保护作用及其机制。方法:培养HUVECs,分为正常组、H2O2模型组、H2O2+OA(0.25μmol/L)组、H2O2+OA(0.50μmol/L)组、H2O2+OA(1.00μmol/L)组、H2O2+维生素C(1.50 mmol/L)组。各组培养24 h后,通过MTT法检测细胞增殖能力,酶生化法测定上清液中的乳酸脱氢酶(lactate dehydrogenase,LDH)和一氧化氮(nitric oxide,NO)含量,酶生化法测定细胞裂解液中的丙二醛(malondialdehyde,MDA)含量、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和超氧化物歧化酶(superoxide dismutase,SOD)活力,流式细胞仪测细胞内活性氧(reactive oxygen species,ROS)和细胞凋亡率。结果:OA能够恢复H2O2损伤的细胞活力,减轻H2O2诱导的LDH释放,减少H2O2诱导的MDA生成,恢复H2O2损伤的GSH-Px和SOD活力,清除H2O2诱导的ROS,减轻H2O2诱导的细胞凋亡,提高HUVECs细胞NO的生成。结论:OA对损伤血管内皮细胞有保护作用,这与OA能够启动HUVECs细胞内抗氧化防御机制,清除细胞内的ROS,提高HUVECs细胞NO的生成,抑制H2O2诱导的HUVECs细胞凋亡有关。

Objective：To study the protective effects and mechanisms of oleanic acid on damaged human umbilical vein endothelial cells（HUVECs） induced by hydrogen peroxide（H2O2）.Methods：HUVECs were divided into control group,H2O2 group（200 %mol/L H2O2 treatment）,H2O2 of 200 %mol/L combined with OA of 0.25 %mol/L treatment group,H2O2 of 200 %mol/L combined with OA of 0.50 %mol/L treatment group,H2O2 of 200 %mol/L combined with OA of 1.00 %mol/L treatment group,H2O2 of 200 %mol/L combined with vitamin C of 1.50 mmol/L treatment group.Cell viability was determined by MTT method.Lactate dehydrogenase（LDH） and nitric oxide（NO） content in the supernatant were determined by biochemical method.The activity of malondialdehyde（MDA）,glutathione peroxidase（GSH-Px） and superoxide dismutase（SOD） in the cell lysate was determined by biochemical method.Intracellular reactive oxygen species（ROS） and cell apoptosis were determined by flow cytometry.Results：OA restored the H2O2-injuried cell viability,reduced the H2O2-induced LDH release,and reduced H2O2-induced MDA formation,recovered H2O2-injuried GSH-Px and SOD activity,removed H2O2-induced ROS,reduced the H2O2-induced apoptosis,and enhanced the NO production in HUVECs.Conclusion：OA has a protective effect on HUVECs,which is related to initiating the antioxidant defense,getting rid of ROS,inhibiting H2O2-induced apoptosis,and improving NO production in HUVECs.