| Objectie:With the rapid development of new type of immunosuppressive and modern medical technology,renal transplantion is becoming more and more useful. and the best way to cure of end-stage renal disease(ESRD)in clinic.It is inevitable that the renal is injured by ischenmia/reperfusion(I/R)in renal transplantion.Serious ischenmia-reperfusion injury is the main reason of delayed graft function(DGF).In clinic,the duration of DGF nearly reflect the degree of ischenmia-reperfusion injury. Lots of clinical and experimental research show,the incidence of acute rejection in patients of DGF is obviously higher than those of renal function recovered in time.Adhesion invasion and activation of leukocyte play an important role in renal ischenmia-reperfusion injury.Leukocyte's accumulation and infiltration in inflammatory sites,is the basic pathology of the inflammatory performance.In recent years,some studies also find that it is an important factor in ischenmia-reperfusion injury,and the adhesion between leukocyte and endothelial cell is basis.After a series of complicated reactions,inflammatory cells adhere to endothelial cells.A large number of experiments show that:leukocyte activation is the first title in renal ischenmia-reperfusion injury.The main mechanism of the injury is that a large number of leukocytes lead to mechanical obstruction in microvascular and limit the blood reperfusion,the activated inflammatory cells release large number of substances,such as:reactive oxygen species(ROS),some of the cytokines and enzymes that can lead to kidney injury;enhanced cell adhesion and activated complement system,these reactions induced to further activation of inflammatory cells and more other cells,therefore it form an gradually enlarged response network,that is inflammatory cascade.Intercellular adhesion molecule-1(ICAM-1) is the major adhesion molecule that mediated leukocyte to adhere to actived endothelial cells.Physiological circumstances,ICAM-1 is expressed in a low level in the kidney. Because oxygen free radicals and cytokines,endotoxin,and thrombin significantly increased,they sustained activated neutrophils and endothelial cells,the ICAM-1 is highly expressed in endothelial.LFA-1 is highly expressed in neutrophil,the construction of LFA-1 Changed at the same time,greatly enhanced the adhesion between LFA-1 and ICAM-1,led to Neutrophils adhere to endothelial cells which highly expressed ICAM-1,led to capillaries narrow,blocking,led to microvascular plug,and no-reflow phenomenon.With the adhesive effect of ICAM-1,Neutrophil cells infiltrated into the surrounding tissue of ischemia-reperfusion,and released more and more inflammatory mediators in local organizations,Such as cytokines, proteases,enzymes and other elastase,Enhanced the adhesion molecule expression and adhesion,aggravated renal injury.Ulinastatin(UTI)is extracted from human urine and refined for a glycoprotein molecular weight of the 6.7KD.It is a broad-spectrum enzyme inhibitors.It significantly inhibited the releasing of inflammatory mediators and stabilized lysosomal membrane,inhibited the releasing of lysosomal,removed free oxygen radicals and the endotoxines in ischenmia-reperfusion injury.Study about UTI show that it inhibits inflammatory cells to infiltrate glomerular,has therapeutic effect on the crescent shape glomerulonephritis.It has protective role on kidney injury caused by various drugs.It has protective role on isolated kidney too.But,the mechanism of the protective effect of UTI on renal ischenmia-reperfusion injury is uncertain.Discussion of ICAM-1 expression in the rat renal of ischenmia-reperfusion injury in different time points and the expression of ICAM-1 in rat renal ischenmia-reperfusion injury which intravenous injected by UTI in different time points.Combination of serum creatinine level,blood urea nitrogen level.and pathological changes in renal tissue.To explore the role of ICAM-1 in renal ischenmia-reperfusion injury.The protective effect and mechanism of UTI on rat kidney ischenmia-reperfusion injury.To provide new ideas for the prevention and treatment of renal ischemia-reperfusion injury in transplantion.Methods:Rat model of ischemia-reperfusion,60 healthy SD rats,weighing 180-200g,male or female,all animals are bought from Animal department of Kunming Medical College,were randomly divided into four groups:sham operation group(C n=15),renal ischenmia-reperfusion group(I n=15),renal ischenmia-reperfusion group with Ulinastatin(I+U n=15),No renal ischenmia-reperfusion with Ulinastatin Group(U n=15).Every group is divided into three groups in different time points(2hour.6hour.12 hour)by executable times.Five rats are conducted to experiment in each time point.Experimental Methods:10%Chloral Hydrate at 3 ml/ kg for intraperitoneal injection,after anesthesia.,Femoral vein for the drugs infusion.UTI 12.5 thousand units dissolved in 1 ml saline;then the U group and the group I+U intravenous inject UTI12.5 thousand units.Group C and Group I intravenous inject saline 1 ml.4 groups rats along the ventral midline laparotomize abdomen and isolate the renal vascula, Right kidney is resected after Renal vascular ligation.I group and I+U group after clipping 60 minutes of left renal artery with non-invasive artery folder,release artery folder to restore blood flow and close abdominal.C group and U group only isolate left renal artery and wait 60 minutes then close abdominal.At this point group U and the group I+U intravenous inject UTI 12.5 thousand units.Group C and Group I intravenous inject saline 1 ml.At the set of ischenmia-reperfusion time point we cut the left kidney and.obtain 5 ml blood from abdominal aorta,Kidneys are fixed in 10%neutral formalin,dehydrated by different levels of alcohol and embedded in transparent paraffin,Sliced into 3um,HE stained,for observation of renal pathology. With immunohistochemical detection method to detect the expression of ICAM-1 in the renal,centrifugal serum from the blood for detection of BUN and Cre. Experimental results of ICAM-1 are analysed by Image Analysis System for semi-quantitative analysis.All data use mean±standard deviation.Group comparison use the multi-factor analysis of variance and each group of data use Pearson correlation data analysis.And we use SPSS16.0 Statistical Analysis Software for data analysis.Results:This study indicates that renal ischenmia-reperfusion injury is obvious, rat renals function were severely damaged by ischenmia-reperfusion,but UTI significantly reduce renal tubular epithelial cells degeneration and necrosis, Significantly improved renal function in each time point after reperfusion,it have protective effect on rat renal.It can inhibit the expression of ICAM-1.Conclusion:First:with the extension of reperfusion time,expression of ICAM-1 is enhanced, pathological injury is aggravated,renal tubular injury score is increased,renal function is also decreased in rat model of ischemia-reperfusion.It is detected that ICAM-1 undoubtedly play an important role in neutrophil activation and the progress of renal ischenmia-reperfusion injury.Second:UTI can significantly inhibit ICAM-1 expression.Reduce rat kidney pathological damage,reduce renal tubular injury score,improve the renal function, we detect that the protective effect of UTI on renal ischenmia-reperfusion injury probably because it inhibit ICAM-1 expression,reduce the inflammation cascadeThird:UTI can improve the renal function after ischemia-reperfusion.It may reduce the DGF incidence,shorten recovery time of the delayed graft function.
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