Ulinastain Protects Human Umbilical Vein Endothelial Cells From Oxidative Stress Via Activation Of Akt/mTOR Pathway

Objective:Ulinastatin, a urinary trypsin inhibitor, has been widely used in perioperative period for its anti-inflammatory properties. However, whether there is other protection mechanism is still not clear. At present study, we investigated the protective effect of ulinastatin on H2O2-induced injury in HUVECs and potential signaling pathway involved.Method:HUVEC cells were divided into5groups:the control group,hydrogen peroxide group, hydrogen peroxide+ulinastatin group, hydrogen peroxide+ulinastatin+LY294002group, hydrogen peroxide+LY294002group. The cell viability were detected by CCK-8method; The generation of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by commercial kit; The level of ROS were determined by flow cytometry; The HUVEC apoptosis rate were detected by flow cytometry; The expression of Akt、Erk、mTOR、caspase3、PARP protein were determined by Western blot.Results:The results indicated that ulinastatin increased the cell viability (34.85±2.67%to68.77%±4.12%)(P<0.01), elevated superoxide dismutase activity (38.81±2.63%to72.77±3.43%)(P<0.01) decreased malondialdehyde formation (338.57±15.50%to138.26±8.37%)(P<0.01) and inhibited reactive oxygen species production (153.46±6.23%to121.57±5.39%)(P<0.01). Furthermore, ulinastatin treatment reversed H2O2-induced apoptosis in HUVECs (18.9±1.37%to7.89±0.12%)(P<0.01), as well as down-regulated PARP and capase-3expression. In addition, these changes were accompany with the activation of PI3K/Akt pathway. However, the anti-oxidative and anti-apoptosis effect of ulinastatin were abolished by LY294002, a phosphatidylinositol-3-kinase (PI3K) inhibitor.Conclusion:Our results suggest the protective effect of UTI against H2O2induced oxidative stress and apoptosis was via activating PI3K/Akt signaling pathway.