| Objective:By studying the expression and change of the NGAL and the KIM-1in the rat model of AKI and cell model with oxidative damage and the patient with AKI,to determine weather the NGAL and KIM-1 in blood and urine could be to predict the happening of ARF and to find reliable biological indicators for early diagnosis of ARF.Method:1.40 SPF male Sprague-Dawley rats randomly assigned to five groups.Four group rats of them were injected introperitoneally with gentamtcin sulfate by 100mg/kg,the control group were injected with saline at the same dosage.One by one group of rats were necropsied respectively on day 3,5,7 and10 of drugs for renal pathological section and immunohistochemistry,serum and urine specimen to detect the change of biochemical index and NGAL,KIM-1.The control group were necropsied two rats each time.To extract the kidney tissue RNA and detect the m RNA expression of every rats in various categories.2.The HK-2 cells were treated with H2O2 at different time points,then the collected supernatant and cells were used to detect the secretion and expression of NGAL and KIM-1 respectively.3.Collected 40 cases blood and urine samples of Clinical patients with acute renal failure during February to December was used to detect the change of biochemical index and the NGAL,KIM-1 in blood and urine.Results:1.The NGAL in blood and NGAL,KIM-1 in urine respectively increased to(28.8±3.17)μg/m L,(3.63±0.50)μg/m L and(.43±1.25)μg/m L after only 3 days, the KIM-1 in blood increased to(4.51±0.41)μg/m L after 5 days.The immunohistochemical results showed that the renal tubular epithelial cells presented dot expression about NGAL after 3 days,the expression of NGAL gradually increased and the staining changed from reddish to dark red-orange with the rats continued to be injected.In addition we could find that the renal tubular wall appeared KIM-1 deposition and the expression of KIM-1 gradually increased and the staining changed from red to dark red-orange with the rats continued to be injected;Serum creatinine and blood urea nitrogen increased to(90.4±1.9)μmol/L,(62.2±1.2)mmol/L after 7 days; cystatin C increased to(0.24±0.03)mg/L after 5 days; the 24 hours microalbuminuria in urine increased to(0.512±0.023)mg/24 h after 8 days;Pathological observation shows renal tubular ectasia, the exfoliation and disappearance of renal tubular wall cells and the inflammatory cell infiltration wew occurred after 5 days.2.With the extension of drug processing time the relative expression of NGAL and KIM-1 and the secretion of NGAL in the cell model with oxidative damage increased,the expression and secretion of NGAL reached the peak 12 hours later with drug treatment and then began to decline after 24 hours;but KIM-1 was n the rise.3.In clinical specimens,NGAL and KIM-1 of blood and urine in the case group were significantly higher than control group.The NGAL and KIM-1 in blood respectively increased to(136.5±18.31)μg/L,(5.21±0.40)μg/L;in urine respectively increased to(28.84±4.06)μg/L,(2.11±0.50)μg/L,while serum creatinine and blood urea nitrogen had not been a significant change.Conclusion:1.A rat model of AKI was successfully constructed,and the rats had typical symptoms of development with AKI.Rat kidney tissue, blood, urine specimens could be detected in a large number of expression of NGAL, KIM-1in the early AKI.Results suggest that both of them are suitable as early diagnostic markers for AKI.2.HK-2 cells what had been induced oxidative damage expressed and secreted large amounts of NGAL, KIM-1 in the process of in vitro culture,it suggests both of them are suitable as the index to Judge the damage of renal epithelial cell.3.The concentrations of NGAL and KIM-1of blood and urine in the case group were significantly higher than that control group,the Cys C was higher slightly,it suggests that the first two could be suitable as the important diagnosis index of AKI. |
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