Effects Of KIM-1on High Glucose Induced The Expression Of MCP-1and FN In Rat Tubular Epithelial Cells

Background and aimsDiabetic nephropathy (DN) is one of the the most common and serious microvascular complications of diabetes mellitus(DM),which is the main reason of end stage renal disease (ESRD) and of the diabetic deaths.Therefore,the prevention and treatment of DN was paid increasing attentions. The main pathological features of DN includes:glomerular hypertrophy,basement membrane thickening, accumulation of extracellular matrix in membrane area, in addition t,renal interstitial fibrosis (RIF) is also important pathological features of progress of DN.The long-term high glucose can cause the renal inherent cells,including the tubular epithelial cells,secreting a variety of inflammatory cytokines and fibrogenic factor, which thereby induce hypertrophy,hyperplasia of cells,increasing extracellular matrix and the occurrence of RIF.Kidney injury factor-1(KIM-1) is a new type I transmembrane protein expressed by varieties of acute and chronic injury renal tubular epithelial cells by,which not only can be used as a an important indicator of the degree of renal injury of acute renal injury (AKI) and chronic kidney disease (CKD),but also has the function involved in injury and repair of the tubular epithelial cell.In recent years,a large number of studies verify that KIM-1closely has a relation to RIF of all kinds of chronic kidney disease,such as DN,but the exact mechanism is unclear.The present studies suggest that KIM-1may be related to renal inflammatory response,and the accumulation of extracellular matrix (ECM) is the main pathological change of RIF.Therefore,we considered high glucose as diabetic conditions,the tubular epithelial cells as the research object in this experiment and observated that effects of KIM-1on high glucose induced the expression of MCP-1and FN in rat tubular epithelial cells,to explore the possible mechanisms of RIFof DN promoted by KIM-1,which thus provides a new theoretical basis for the treatment of DN.MethodsThe tubular epithelial cells was cultured in vitro and divided into several groupes: control group (D-glucose5.6mmol/L);hypertonic group (D-glucose5.6mmol/L+D-mannitol24.4mmol/L);high glucose group (D-glucose30mmol/L);Control siRNA group;KIM-1siRNA group.Enzyme-linked immunosorbent assay (ELISA) was used to assess the protein expression of MCP-1and FN in the cells supernatant;Western blot was used to detect the protein expression of KIM-1;reverse transcription-polymerase chain reaction (RT-PCR) detected mRNA expression of KIM-1,MCP-1and FN.Results1.The control group and hypertonic group almost had no the protein and mRNA expression of KIM-1that were increased at12h in the high glucose group,compared with the control group (P＜0.05),and reached the peak at48h (P＜0.05).2.Compared with the control group,the protein and mRNA expression of MCP-1and FN in high glucose group were increased at24h significantly (P＜0.05),and peaked at48h (P＜0.05).3.Compared with the high glucose,the protein and mRNA expression of MCP-1and FN were decreased in KEM-1siRNA group (P＜0.05).ConclusionHigh glucose can induce expresssion of KIM-1,MCP-1and FN of the tubular epithelial cells and down-regulating KIM-1can significantly inhibit the expression of MCP-1and FN,which suggests that KIM-1may be involved in RIF of DN through regulating expression of MCP-1and FN.