榆干离褶伞溶栓酶对过氧化氢损伤血管内皮细胞的保护机制研究

Protective effects of fibrinolytic enzyme from Lyophyllum ulmarium on vascular endothelial cells injury induced by H_2O_2

目的:研究榆干离褶伞(Lyophyllum ulmarium,LU)溶栓酶对氧化应激所致损伤血管内皮细胞的保护机制研究。方法:以过氧化氢诱导人脐静脉内皮细胞(HUVEC)的氧化损伤,通过甲基噻唑基四唑(MTT)法和细胞培养上清液的乳酸脱氢酶(LDH)活性来分析细胞损伤程度;ELISA方法测定肿瘤坏死因子α(TNF-α)水平;硝酸还原酶法测定细胞NO含量;Western印记法检测NF-κB p65、iNOS和eNOS蛋白表达。结果:LU溶栓酶预处理后显著提高细胞存活率,抑制过氧化氢所致的LDH、TNF-α和NO水平的升高,同时抑制NF-κB、p-NF-κB、iNOS和eNOS的表达。结论:LU溶栓酶对血管内皮细胞的保护作用是通过下调NF-κB信号通路来实现的。

Objective： To observe the protective effects of fibrinolytic enzyme from Lyophyllum ulmarium（Lu） on H_2O_2-induced oxidative damage of human umbilical vein endothelial cells（HUVECs）. Methods： The H_2O_2 was used to induce the oxidative injury model of HUVECs. The cell viability and injury were assayed by MTT assay and LDH leakage. The TNF-α level was detected by ELISA method and the NO level was measured by nitrate reductase method. The expression of NF-κB p65, iNOS and eNOS proteins was detected by western blot. Results： Lu could increase the survival rate of endothelial cells, and downregulate the expression of NF-κB p65, p-NF-κB, iNOS and eNOS proteins. Conclusions： The protective effects of fibrinolytic enzyme from Lyophyllum ulmarium on vascular endothelial cells might be due to down-regulation of the NF-κB signaling pathway.