榆干离褶伞溶栓酶调控Nrf2/Sirt3、Nrf2/ARE信号通路改善内皮细胞氧化损伤OA

To explore the protective effect of L.ulmarium fibrinolytic enzyme(LUFE)against oxidative damage in human umbilical vein endothelial cell(HUVEC),low(40 μg·mL-1)and high(80 μg·mL-1)doses of LUFE were used to treat an oxidative stress injury model induced by H2O2.Then,various parameters were measured,including cell mortality,the activities of superoxide dismutase 2(SOD2),glutathione peroxidase(GSH-Px),and catalase(CAT),the levels of malonaldehyde(MDA),adenosine triphosphate(ATP),mitochondrial membrane potential,cellular reactive oxygen species(ROS),mitochondrial ROS,and the protein expression levels of Kelch-like ECH-associated protein 1(Keap-1),nuclear factor erythroid 2-related factor 2(Nrf2),silent information regulator 3(Sirt3),NAD(P)H-quinone oxidoreductase 1(NQO1),heme oxygenase 1(HO-1),Bcl2-associated X protein(Bax),B cell lymphoma/leukemia-2(Bcl2),cleaved-cysteinyl aspartate-specific proteinase 3(cleaved-caspase 3),and cleaved-cysteine aspartic acid-specific protease 9(cleaved-caspase 9).The results showed that LUFE decreased the H2O2-induced apoptosis rate,decreased both cellular mitochondrial ROS levels,increased mitochondrial transmembrane potential,enhanced the activities of SOD2,CAT,GSH-Px,elevated ATP content,and reduced MDA content.LUFE upregulated the ratio of Bcl2 to Bax protein expression levels(Bcl2/Bax)and the protein expression levels of Nrf2,HO-1,NQO1,Sirt3.In contrast,it downregulated the protein expression levels of cleaved-caspase 9,cleaved-caspase 3 and Keap-1.These findings suggested that LUFE alleviates H2O2-induced oxidative damage and reduces apoptosis in endothelial cells by regulating the Nrf2/Sirt3 and Nrf2/ARE signaling pathways.