The Protective Effect Of Hepatocyte Growth Factor Naked DNA Transfer By Hydrodynamics Injection On Renal Ischemia-reperfusion Injury

Objective:Ischemia-reperfusion injury(IRI) is the cause of acute renal failure(ARF). Ischemia is inevitable in renal transplantation,ischemia-reperfusion injury-induced acute renal failure is closely related to chronic renal injury and long-term graft failure Many experimental results indicates that hapatocyte growth factor(HGF) in the prevention and treatment of renal ischemia-reperfusion injury has good application prospects.However,exogenous HGF is extremely unstable,due to the rapid metabolism of the liver,half-life of HGF in the blood circulation only 3～5min, so injection of HGF can not achieve an effective therapeutic effect.The purpose of this experiment is to find an efficient exogenous HGF DNA transfection methods,to lay a foundation for the clinical application of HGF.Method:1.To 40 SD rats were randomly divided into 4 groups,each group with 10.They are,A:Injected hepatocyte growth factor naked plasmid HGF DNA into the renal vein, then established IRI model of SD rats;B:Established IRI model of SD rats and injected hepatocyte growth factor naked plasmid DNA into the renal vein the same time;C:Injected hepatocyte growth factor naked plasmid DNA into the femoral vein, then established IRI model of SD rats;D:Injected normal sodium into the renal vein, then established IRI model of SD rats.Compared the effect of naked plasmid HGF DNA transfection and the efficiency of HGF DNA transfection in different ways.2.After 24h and 48h of the establishment of IRI models,respectively,resected the left kidney and collected Venous blood.3.Detect serum crea,urea nitrogen and HGF.4.To take left part nephridial tissue do hematoxylin and eosin dyeing.Under common practice pathology checking and under Paller's nePhric tubule score that definite the degree of injury in Pathology in each group.Detct the Bax,Bcl-2,HGF of nephridial tissue and apoptosis by immunohistochemistry methods.Observe ultrastructure of renal tubular by electron microscopy.Result:The value of serum crea,urea nitrogen and the tatio of apoptosisEach group of serum urea nitrogen and serum creatinine at different time points (24h,48h) as compared with the saline group were significantly different.(P<0.05). And renal function index among the three groups compared to each other are significantly different,the renal injury of renal vein HGF DNA transfection group was the lightest,and among the rest of the groups,the extent of renal injury from mild to severe is:IRI followed by HGF DNA transfection at the same time group,femoral vein HGF DNA transfection group and saline control group.Apoptosis rate(24h,48h) from low to high is:renal vein HGF DNA transfection group,IRI followed by HGF DNA transfection at the same time group,femoral vein HGF DNA transfection group and saline control group.By the statistical software analysis,each group at the same time point between apoptosis rate and corresponding serum creatinine,blood urea nitrogen values were the positive correlations(P<0.01).2.The value of Bax,Bcl-2 and Bcl-2/BaxEach group(24h,48h)of Bax content from low to high is:renal vein HGF DNA transfection group,IRI followed by HGF DNA transfection at the same time group, femoral vein HGF DNA transfection group and saline control group,and the value of Bcl-2 was just opposite.The value of Bcl-2/Bax from high to low is:renal vein HGF DNA transfection group,IRI followed by HGF DNA transfection at the same time group,femoral vein HGF DNA transfection group and saline control group,The above-mentioned set of data analyzed by statistical software,the results are statistically significant and significantly different(P<0.01).3.HGF content in renal tissueDetected by immunohistochemistry,in each group(24h,48h),the HGF levels in renal tissue when compared with the saline control group,with Statistically significant differences in test.HGF content in each group followed by descending is:renal vein HGF DNA transfection group,IRI followed by HGF DNA transfection at the same time group,femoral vein HGF DNA transfection group and saline control group.There are significant differences after statistically test.(P<0.01)4.The degree of renal tubular injuryRat renal tissue stained by HE,the optical microscope(×200) on the extent of renal tubular injury score,each group(24h,48h) as compared with the saline control group,the degree of renal tubular injury score were lower than the latter,there are significant differences after the statistically test.Between apoptosis rate and renal tubular score,there is a positive correlation after statistical analysis.(P<0.01)5.Content of serum HGFThe two time periods(24h,48h) in each group,the concentrations of HGF serum compared with the saline control group were significantly different(P<0.05), compared among the three groups are also significant differences(P<0.05).Content of serum HGF from high to low is:IRI followed by HGF DNA transfection at the same time group,femoral vein HGF DNA transfection group,renal vein HGF DNA transfection group and saline control group.Conclusion:1.Both of the two ways,the the naked HGF DNA plasmid injected into the femoral vein or the renal vein can significantly reduce the renal ischemia-reperfusion injury.2.Target organ-renal vein naked plasmid HGF DNA transfection transfection than the femoral vein can reduce ischemia-reperfusion injury,and the effect of transfection that naked plasmid before ischemia is more visible than that IRI followed by HGF DNA transfection at the same time.