Theprotectiveeffectoflyophyllumulamariumstr Eprokinaseinvascularendothelialcells

Objective:Research on the protective effects of Lyophyllum ulamarium streptokinase in endothelial cells and its mechanism.Methods:(1) cellular experiments:MTT assay was used to determine the effects of Lyophyllum ulamarium streptokinase on viability in HUVEC which was established from stimulating by H2O2. Cells and medium were collected to detect the effects of Lyophyllum ulamarium streptokinase on H2O2-induced apoptosis in HUVEC. Flow cytometry was performed to exam mitochondrial transmembrane potential (△Ψm) and ROS. Western Blot was used to exam the protein levels of NF-κB-p65, iNOS, eNOS, Caspase-8, Caspase-9, Caspase-3. NO level was detected by the method of nitric acid reductase.(2) Animal experiments:Stress injury were formed by Adrenaline and Ice water stimulation. Detection the number of circulating endothelial cells in plasma. Elisa assay was used to detect AT-Ⅲ activity.Results:(1) cell experiments:MTT results showed that within a certain range of concentrations, Lyophyllum ulamarium streptokinase take a protective effect in H2O2stimulated HUVEC; flow cytometry results showed that compared with the normal group, model group had a significant reduce in mitochondrial transmembrane potential, increase in intracellular reactive oxygen species. Compared with model group, treatment group had a significant increase in mitochondrial transmembrane potential, reduced in intracellular reactive oxygen species. Western bolt showed that compared with the normal group, model group can significantly increase the NF-κB-p65protein expression, while increase iNOS, eNOS expression, and increased intracellular NO. Compared with the model group, the treatment group can significantly reduce the NF-κB-p65protein expression, while reduce iNOS, eNOS expression, and reduce intracellular NO.(2) In animal experiments:Rat model of endothelial cell disorder results show that compared with the normal group, the number of circulating endothelial cells increased in model group, and AT-Ⅲ activity were reduced. compared to model group, the number of circulating endothelial cells reduced in treatment group, and AT-Ⅲ activity were increased.Conclusion:Lyophyllum ulamarium streptokinase have a protective effect in vascular endothelial cells. It also had protective effects on antioxidant effects. Lyophyllum ulamarium streptokinase have a protective effect in endothelial cells, which may be related to NF-κB-iNOS-NO pathway regulation. Lyophyllum ulamarium streptokinase able to promote the secretion of AT-Ⅲ in vascular endothelial cells.