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目的:研究蛇毒三肽pENW对体外过氧化氢诱导人脐静脉内皮细胞损伤的保护作用。方法:用体外培养的人脐静脉内皮细胞株传代后进行实验,H2O2(300μmol/L,12h)模拟氧化损伤模型;药物处理组分别加入不同剂量的蛇毒三肽pENW(10-4mol/L,10-5mol/L,10-6mol/L)以及阳性对照药依达拉奉(10-5mol/L);用噻唑蓝(MTT)比色法检测细胞活力;比色法检测细胞培养上清液中乳酸脱氢酶(LDH)的漏出率,细胞中超氧化物歧化酶(SOD)的活性和谷胱甘肽(GSH-Px)的含量;Annexin V-FITC/PI双染法流式细胞仪检测细胞凋亡。结果:蛇毒三肽pENW(10-4mol/L,10-5mol/L)能够显著抑制H2O2对人脐静脉内皮细胞的氧化损伤作用,降低H2O2诱导人脐静脉内皮细胞的凋亡,降低LDH漏出率,提高人脐静脉内皮细胞中SOD、GSH-Px活性。结论:蛇毒三肽pENW可降低过氧化氢对人脐静脉内皮细胞的氧化损伤作用,其保护作用可能与抗氧化及抑制细胞凋亡有关。
Objective: To study the protective effect of snake venom tripeptide pENW on H2O2-induced injury of human umbilical vein endothelial cells in vitro. Methods: Human umbilical vein endothelial cell lines cultured in vitro were passaged, and the model of H2O2 (300μmol / L, 12h) was used to model the oxidative injury. The drug-treated groups were treated with different doses of venom tripeptide pENW (10-4mol / L, 10 -5mol / L, 10-6mol / L), and positive control drug (10-5mol / L). Cell viability was measured by MTT colorimetric assay. The leakage rate of lactate dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and the content of glutathione (GSH-Px) in the cells were detected by flow cytometry; Annexin V-FITC / Apoptosis. Results: Snake venom tripeptide pENW (10-4mol / L, 10-5mol / L) could significantly inhibit H2O2-induced oxidative injury of human umbilical vein endothelial cells and reduce H2O2-induced apoptosis of human umbilical vein endothelial cells and reduce LDH leakage rate , Improve the activity of SOD, GSH-Px in human umbilical vein endothelial cells. CONCLUSION: Snake venom tripeptide pENW can reduce the oxidative damage of H2O2 on human umbilical vein endothelial cells, and its protective effect may be related to the anti-oxidation and the inhibition of apoptosis.