RESUMO
Objective To evaluate the effects of KIM-1 on high glucose induced the expression of MCP-1 and FN in rat tubular epithelial cells and to explore the possible mechanisms of KIM-1 involved in renal interstitial fibrosis of DN.Methods The rat renal tubular epithelial cells (NRK52E) were cultured in vitro and divided into five groups:Normal control group (D-glucose 5.6 mmol/L),Hypertonic group (D-glucose 5.6 mmol/L + D-mannitol 24.4 mmol/L),High glucose group (Dglucose 30 rmmol/L),Control siRNA group,KIM-1 siRNA group.ELISA assay was used to assess the levels of MCP-1 and FN in the cells supernatant; Western blotting was used to detect the protein expression of KIM-1; RT-PCR was used to detect mRNA expression of KIM-1,MCP-1 and FN.Results Compared with the control group,the protein and mRNA expression of KIM-1 in the high glucose group were increased at 12 h (P ＜ 0.05),and reached the peak at 48 h (P ＜ 0.05); the protein and mRNA expression of MCP-1 and FN in high glucose group were increased at 24 h significantly (P ＜ 0.05),and peaked at 48 h (P ＜ 0.05).Compared with the high glucose group,the protein and mRNA expressions of MCP-1 and FN in KIM-1 siRNA group were decreased (P＜0.05).Conclusions Down-regulating the expression of KIM-1 can significantly inhibit the expression of MCP-1 and FN,which suggests that KIM-1 may be involved in renal interstitial fibrosis of DN by regulating expression of MCP-1 and FN.
