Protective Effect Of Oleuropein On HK-2 Cells Injured By Oxygen-Glucose Deprivation

Objective:To investigate the protective effect and mechanism of oleuropein on HK-2 cells injured by oxygen-glucose deprivation(OGD).Methods:The HK-2 cells was purchased from the American type culture collection.After the arrival of the cells,we observe the shape,density and whether were polluted.Then,the cells were placed in the incubator 4-6h that we insure the normal of the cells.Preparation of cell hypoxia re-oxygenation model:The three gas incubator was adjusted by the ventilation ratio of 95%N2 and 5%CO2 to simulate the anoxic environment in vitro for the treatment of HK-2 cells.After 24 hours of anoxia and different times of re-oxygenation,the cell phenotype was observed by optical microscope to clarify the preparation conditions of the model.HK-2 cells were treated with different doses of oleuropein.In each group,A450 values in cells were assayed with CCK-8(Cell Counting Kit-8)test,Afterward the apoptosis of HK-2 cells were evaluated by fluorescence microscope after Hoechst staining,The expression of Bcl-2,Bax protein and were detected by Western blot.Results:1.cells were anoxic for 24h,and after re-oxygenation at different time,the cell phenotype showed a certain time dependence.However,hypoxia 24h re-oxygenation 3h is sufficient to simulate renal ischemia-reperfusion injury in vitro.2.CCK-8 test results show:In contrast with the control group,Oleuropein could increase the survival rate of HK-2 cells.3.Apoptosis related changes were observed after Hoechst33258 staining,including nuclear condensation,chromatin condensation and apoptotic body formation and presented in a concentration dependent manner.This showed oleuropein can reduce cell apoptosis caused by OGD.4.Blotting showed that,Oleuropein could also increase the expression of Bcl-2 protein and decrease the expression of Bax protein,and the difference was statistically significant(P<0.05).Conelusion:Oleuropein could relieve the HK-2 cell injury induced by OGD and the protective effect may be related to inhibiting cell apoptosis.