Effect Of EPO On High Glucose-Cultured Apoptosis Of Human Glomerular Mesangial Cells And Its Possible Mechanisms

ObjectiveTo explore the effect of erythropoietin?EPO?on high glucose-induced apoptosis in human glomerular mesangial cells and its possible mechanisms of action,and to find new treatments for delaying the progress of diabetic nephropathy?DN?.MethodsThe cultured human glomerular mesangial cells were randomly divided into 8groups:control group?group A,cells received no treatment?,high glucose group(group B,cells were treated with 30 mmol·L^-1D-glucose),high permeability group?group C,cells were treated with 24.5 mmol·L^-11 mannitol?,EPO group?group D,cells were treated with 20 U/ml EPO?,low-concentration EPO intervention group(group E,cells were treated with 5 U/ml EPO+30 mmol·L^-1D-glucose),middle-concentration EPO intervention group(group F,cells were treated with 10 U/ml EPO+30 mmol·L^-1D-glucose),high-concentration EPO intervention group(group G,cells were treated with20/ml EPO+30 mmol·L^-1D-glucose),and Y27632 group(group H,cells were treated with30 mmol·L^-1D-glucose+10 u mol·L^-1Rho-kinase inhibitor).Cells were cultured for 24 hours.The mRNA expression of RhoA and ROCK was detected by RT-PCR.The apoptosis of cells was measured by flow cytometry.ResultsThe apoptosis rate of cells in group A?control group?was not different from that in group C?high permeability group?,D?EPO group?or E?low-concentration EPO intervention group??P>0.05?.Compared with group B?high glucose group?,the apoptosis rate and expression of RhoA and ROCK decreased in groups C?high permeability group?,D?EPO group?,E?low-concentration EPO intervention group?,F?middle-concentration EPO intervention group?and G?high-concentration EPO intervention group?,but increased in group H?Y27632 group??P<0.05?.Compared with group E?low-concentration EPO intervention group?,the apoptosis rate increased but the expression of RhoA and ROCK decreased in groups F?middle-concentration EPO intervention group?,G?high-concentration EPO intervention group?and H?Y27632 group??P<0.05?.Compared with group F?middle-concentration EPO intervention group?,the apoptosis rate and ROCK1 expression reduced in groups G?high-concentration EPO intervention group?and H?Y27632 group?,RhoA expression decreased in group G?high-concentration EPO intervention group?,and RhoA expressionin creased in group H?Y27632 group??P<0.05?.Pearson analysis showed a positive correlation between RhoA and ROCK1 expression in groups B?high glucose group?,E?low-concentration EPO intervention group?,F?middle-concentration EPO intervention group?and G?high-concentration EPO intervention group??r=0.829,0.831and0.861,respectively;P<0.05?.Conclusion1.High glucose environment can inhibit the growth of normal human mesangial cells.and its mechanism may be due to the activation of RhoA/ROCK signaling pathway in high glucose environment to promote the apoptosis of mesangial cells;2.At the level of human erythropoietin provided by the experiment,human erythropoietin inhibits the apoptosis of normal human mesangial cells,suggesting that it has the effect of protecting the kidney,and the inhibition is related to human erythropoietin.Relevant to the concentration level.3.The protective effect of human erythropoietin on normal human mesangial cells in high glucose environment may be related to blocking RhoA/ROCK signaling pathway.