RhoA/ROCK Signaling Pathway In The Role Of Albumin Induced HK-2Cells Transdifferentiation And RHuEPO Intervention Effects

Purpose:This paper aims to explore research the role of RhoA/ROCKsignaling pathway in the action of human albumin-induced HK-2celltransdifferent-iation.and rHuEPO intervention effect.Looking to explore the possiblemechanism of renal protective effect of rHuEPO.Method:The vitro cultured HK-2cells were randomized:the control group (anyiants), E1group (no stimulus), E2group (join20U/mL rHuEPO), E3group (join5mg/mL albumin), E4group (5mg/mL albumin+5U/mL rHuEPO), E5group(5mg/mL albumin+10U/mL rHuEPO), E6group (5mg/mL albumin+20U/mLrHuEPO), E7group (join5mg/mL albumin after10μmol/L Y27632for30min),acting on each group for24h. RT-PCR to detect the expression the levels ofRhoA, ROCK1mRNA,Immunofluorescence to detect the cell E-cadherin (E-cadherin)and α-smooth muscle actin (α-SMA) protein expression,ELISA test the supernatantprotein fibronectin (FN).Results: E1and E2group display that HK-2cells showed morphological ofcobblestone,and giving5mg/mL albumin stimulated that elongated fusiform cellsexhibit changed, showing fibroblast-like appearance; rHuEPO in5U/mL,10U/mL,20U/mL can prevent albumin concentration-dependent HK-2cells induced fibrosis,cobblestone-like cells to re-turn; E7group were observed in the visible oval cellmorphology, cell gap slightly increased, but the cell degree of change smaller than thegroup E3.RT-PCR results showed that albumin induced group the expression of RhoA,ROCK1mRNAwas significantly higher than the control grou（pP<0.05）, significantlyreduce the expression of RhoA, ROCK1mRNA after rHuEPO given intervention（differences between groups P<0.05）,ELISAand immunofluorescence showed thatrHuEPO for HK-2cells EMT has prevented action,has dose-dependent manner, thegreater the concentration, the more obvious blocked.Conclusion: Albumincan activate the renal tissue RhoA/ROCK pathwayinvolved in the process of transdifferentiation, mainly through improving the levels ofHK-2cells, RhoA, ROCK1mRNA, increasing the intracellular levels of α-SMA accompanied by reduced E-cadherin,extracellular matrix FN protein increased.rHuEPO can dose-dependent manner to prevent albumin induced HK-2cellstransdifferentiation process, parts may be inhibited byRhoA/ROCK pathway activadeffects, final to achieve the purpose of delaying renal fibrosis.