Effect Of Erythropoietin On High Glucose-induced Fibrosis Of Human Kidney Mesangial Cells(MCs) And Its Possible Mechanism

Objective:This study aimed to observation the proliferation effect of rythropoietin(EPO) on high glucose-induced fibrosis of human kidney mesangial cells(MCs) and expression of cytokines and inflammatory factors, investigating the biological effects of EPO on renal tissue after application may arise. Methods:During December 2014- May 2015, MCs cultured in vitro were randomly divided into a bland control group, a 30 mmol/L high glucose induction group, 24.5 mmol/L amannitol induction group, a 20 U/ml EPO induction group, an EPO inhibition group(5,10 and 20 U/ml+30 mmol/L high glucose), and an Rho kinase inhibitor group(30 ?mol/L Y27632 + 30 mmol/L high glucose). After 24 hours of intervention, the mRNA levels of RhoA and ROCK were determined by RT-PCR; Cell proliferation was measured by CCK-8; Those of Connective tissue growth factor(CTGF),Collagen IV?Interleukin-6(IL-6) and Tumor necrosis factor-?(TNF-?) in supernatant were detected by ELISA. Results:Compared with the blank control group, the expressions of RhoA and ROCK1 mRNA were significantly increased in the high glucose induction group(P<0.05),but markedly deceased in the 5,10,20 U/ml EPO inhibition groups and Rho kinase inhibitor group(P<0.05), there was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in EPO group. Compared with the blank control group, the expression of CEGF?Collagen IV?IL-6 and TNF-? were increased in the high glucose induction group(P<0.05), but markedly deceased in the 5,10,20 U/ml EPO inhibition groups and Rho kinase inhibitor group(P<0.05),and the effects were also related to the concentration of EPO. CCK-8 method showed that EPO significantly promoted the proliferation of MCs(P<0.05), after high glucose can inhibit proliferation in normal human MCs, adding different concentrations of EPO or Y27632 intervention, proliferation was significantly inhibited(P<0.05),and EPOpromoted proliferation in a concentration-dependent manner Conclusion:High glucose inhibited the MCs proliferation;EPO concentration in a certain time and can stimulate the proliferation of MCs cultured in high glucose, inhibiting the expression of MCs' cultured in high glucose CTGF, IV collagen protein and IL-6 and TNF-?, and protect the kidneys from renal interstitial fibrosis, which may be related Rho A / ROCK signaling pathway.