The Effects Of Injured Endothelial Cells On Hypoxic Injury Of Renal Tubular Cells

Objective:By analyzing the injury factors of epithelial cell in HK-2 in each group,the effects of injured endothelial cells on hypoxic injury of renal tubular cells were discussed,and the role of endothelial cell in acute ischemia-reperfusion induced acute kidney injure was inferred.Methods:DMEM/F12 culture medium containing 20% fetal bovine serum was used for culturing HK-2 and M199 culture medium containing 20% fetal bovine serum,1% penicillin-streptomycin solution,0.03g/l ECGS,5ug/ml heparin,10 mM L-glutamine was used for culturing HUVEC,they were respectively cultured in a constant temperature incubator at 37? and 5%CO2,and were passed according to the cell growth density ratio of 1:3.Experimental groups: HK-2 mono-cultured in a normoxic condition(N0);HK-2 cultured in a hypoxic condition(H0);HK-2 co-cultured with normoxic HUVEC in a hypoxic condition(H1);HK-2 co-cultured with pre-hypoxic injured HUVEC in a hypoxic condition(H2);E-cadherin and iNOS of HK-2 in each group were detected by using western blot;the growth of HK-2 in each group was observed by Hematein Eosin staining.Results:Immunofluorescence showed that after culturing in hypoxia for 24 hours HIF-1?passed into the nucleus of HUVEC;the results of Western Blot showed that compared with the group N0,the expression of E-cadherin protein in the H0 and H2 were significantly reduced,the decrease in the H0 group was about 40% of that in the N0(p<0.05),and the decrease in the H2 was about 90% of that in the N0(p<0.05),while the expression of E-cadherin in the H1 was increased by about 30% of that in the N0(p<0.05);however,the expression of iNOS protein was increased in the H0 and H2,the increase of H2 was more significant,the increase of iNOS protein in the H0 was about 200% compared with that in the N0(p<0.05);the increase of iNOS in the H2 was about 40% compared with that in the H0(p<0.05).The number of suspended cells in H0 and H2 was significantly higher than that in N0 and H1.HE staining showed that in the N0 the cytoplasm and cell boundary of HK-2 were clearly showed;The cell structured of HK-2 in the H0 was disordered and the cell boundary was not clear;The cell morphology of HK-2 in the H1 was relatively complete;The cell structure of HK-2 in the H2 was disordered,the boundary of nucleus and cytoplasm was hard to be seen,in some where it was broken,and the nucleus of that cell even disappeared Conclusion:Pre-hypoxic injured HUVEC cells can exacerbate hypoxic injury of HK2 cells by releasing some substances,while,resting HUVEC cells can release some substances which protect HK-2 cells from being injured by hypoxia.