| Text:Uremia is the terminal stage of chronic renal failure,which incidence is 0.01% every year in china,most of patients are adults.It Seriously affected peoples'health and safety.Kidney transplantation is the best treatment of terminal stage of chronic renal failure,which can significantly improve the patients'quality of life,reduce complications and mortality rate.Although perioperative complications and early acute rejection rate was significantly lower and graft survival in one year was greatly increased because of the improvement in surgical techniques and the application in new immunosuppressive drugs,but, half-lifetime of graft did not significantly improve.So there are more and more attention in chronic allograft nephropathy.Cold ischemia reperfusion injury is the stage of inevitable pathophysiology in clinical organ transplantation and one of risk factors in chronic allograft nephropathy.Grafts'cold ischemia time is the main reason of delayed graft function after renal transplantation.delayed graft function and acute rejection episodes are the crucial factors of graft survival time.Recently studies discovered that the inflammation and immune response involved in ischemia-reperfusion injury. Inflammatory cell infiltration,cytokine expression upregulation and T cell mediated immune response in ischemical reperfusion injury process cause the kidney damage.Immune system Is the complex mechanism for defense which gradually evolve in the long-term in human's body, including the innate and acquired immunity. It is widely considered recently that innate immunity involve in the activation of acquired immunity. Especially the family of TLRs, belong to innate immunity,the receptor of innate immunity.TLRs identified PAMP like bacteria, viruses,fungi and protozoa,and identify endogenous ligand release by damage or necrotic cells damaged cells caused of a variety of factors like HMGB1,and then triggered a series of inflammatory response.HMGB1 is a class of nonhistone chromatin protein which hus been found in 30 years ago,belong to the family of high mobility group protein.It is think that HMGB1 is a kind of nuclear protein in the earlier study,which interaction with DNA in the nucleus such as transcription, replication, recombination and other complex process of orderly function.Recently studies discovered that HMGB1 might have the activity in "early" and "late"mediators of inflammation,and play the important role in various types of acute and chronic inflammatory diseases,such as arthritis,sepsis,trauma, acute lung injury, ischemia reperfusion injury.HMGB1 research has focused on the model of warm ischemia-reperfusion in rats in domestic and foreign research recently.The studies show that endogenous ligand (HMGB1) combined with the TLRs can induce dendritic cells (DC) maturation, In addition to production of cytokines and chemical factors, mature DC surface costimulatory molecules can provide a "second signal" to T lymphocytes.binding of TLRs and ligand can trigger effect by T helper cell (TH cells) in DC.Conversely, if the lack of TLRs of exogenous and endogenous ligands, TLRs can not be activated, the DC mature obstacles, and immature DC can not start the T lymphocyte response. The injury mechanism and characteristics of renal cold ischemia and reperfusion is not clear.Experimental study, blocking the activity of extracellular HMGB1,has become an important intervention strategies related to the disease process.Ethyl pyruvate is the food additive classified as non-toxic substances by U.S.Food and Drug Administration, has the significant affect of inhibition in the synthesis and release of the HMGB1,and protective effect on Ischemia-reperfusion injury.It has great clinical value in further study of EP.By study of HMGB1 in different model of renal ischemia-reperfusion in the expression and application of EP to intervention,The topic are discuss the following issues:1.Construction of renal warm ischemia-reperfusion injury and cold ischemia-reperfusion injury,Observe the different time points after ischemia HMGB1, inflammatory factor NF-κB,TNFαexpression and renal injury.2.Approach the mechanism of the acquired immune of HMGB1 start of renal heat, cold reperfusion injury.3.Observe the effect of EP treat the warm ischemia reperfusion injury, and accordingly treat cold ischemia and reperfusion injury, provide prevention the immunological treatment of renal cold ischemia-reperfusion injury. In the current case of shortage of organ donors. How to maintain the organ cold ischemia time longer shelf, Expand the use of donor,reduce the incidence of delayed graft function and chronic allograft nephropathy caused by graft cold ischemia time after transplantation, extended half-lifetime of transplanted kidney.It has the important research significance.Chapters 1:The Study of Expression and Intervention of HMGB1 in the Warm Ischemia Reperfusion Injury in KidneyObject:To approach the path of HMGB1 induced warm renal ischemia-reperfusion injury, identify the expression of HMGB1 in warm ischemia-reperfusion injury in kidney, apply the EP to intervention, identify the effect of treatment on EP. Methods:Sixty SD rats were randomly divided into three groups:sham group (n=20), warm ischemia-reperfusion injury group(n=20), and treatment group(n=20). The animal model of ischemi-reperfusion injury was prepared by clamping left renal pedicle of kidney for 45 minutes,then the right kidney was resected after kidney reperfusion. The animals in cold ischemi-reperfusion injury group were given 4 ml Ringer's solution and in treatment group were given Ringer's EP solution(EP was diluted in Ringer's solution, and the concentration was 3.26 g/L) at 20 minutes before the ischemia, and again immediately after reperfusion by intravenous injection via dorsal penile vein. The blood samples from inferior vena cava were collected at 6hour,1day,3day,5day after the establishment of model respectively,and the animals were sacrificed to get kidney tissues.The changes of serum levels of creatinine(CRE),urea nitrogen(BUN), the contents of HMGB1,nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)as well as tumor necrosis factorα(TNF-α)in kidney tissues, immunohistochemical expression of kidney were determined at various time points. Result:HMGB1 levels in warm ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group. Among these, warm ischemia-reperfusion injury group was higher than treatment group.serum CRE as well as BUN levels in warm ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group. Among these,warm ischemia-reperfusion injury group was higher than treatment group. NF-κB as well as TNF-alevels in warm ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group. Among these, warm ischemia-reperfusion injury group was higher than treatment group. The damage of kidney tissues in warm ischemia-reperfusion injury group and treatment group were more severe than those in sham group. Among these, warm ischemia-reperfusion injury group was more severe than treatment group.Conclusion:It was observed that HMGB1 detected in each group in kidney specimens. the expression of HMGB1 increased after reanl warm ischemia-reperfusion injury,lead to the expression of CRE,BUN,TNF-α,NF-κB increased,application of EP to block HMGB1 can reduce the expression of HMGB1, and then reduce the reduce the expression of CRE,BUN,TNF-α,NF-κB.EP help to reduce the damage caused by renal warm ischemia-reperfusion injury.Chapters 2:The Study of Expression and Intervention of HMGB1 in the Cold Ischemia Reperfusion Injury in KidneyObject:To approach the path of HMGB1 induced cold renal ischemia-reperfusion injury, identify the expression of HMGB1 in cold ischemia-reperfusion injury in kidney.According to the result of warm ischemia-reperfusion injury test, Apply the EP to intervention, identify the effect of treatment on EP.Methods:sixty SD rats were randomly divided into three groups:sham group(n=20), cold ischemia-reperfusion injury group(n=20), and treatment group(n=20).Construction the animal model of cold ischemia-reperfusion injury.The animals in cold ischemi-reperfusion injury group were given 4 ml Ringer's solution and in treatment group were given Ringer's EP solution(EP was diluted in Ringer's solution, and the concentration was 3.26 g/L) at 20 minutes before the ischemia, and again immediately after reperfusion by intravenous injection via dorsal penile vein. The blood samples from inferior vena cava were collected at 6hour,1day,3day,5day after the establishment of model respectively, and the animals were sacrificed to get kidney tissues. The changes of serum levels of creatinine (CRE),urea nitrogen (BUN),the contents of HMGB1,nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) as well as tumor necrosis factor a (TNF-a)in kidney tissues immunohistochemical expression of kidney were determined at various time points. Result:HMGB1 levels in cold ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group. Among these, cold ischemia-reperfusion injury group was higher than treatment group.serum CRE as well as BUN levels in cold ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group.Among these, cold ischemia-reperfusion injury group was higher than treatment group. NF-κB as well as TNF-alevels in cold ischemia-reperfusion injury group and treatment group were significantly higher than those in sham group.Among these, cold ischemia-reperfusion injury group was higher than treatment group. The damage of kidney tissues in cold ischemia-reperfusion injury group and treatment group were more severe than those in sham group. Among these,coldm ischemia-reperfusion injury group was more severe than treatment group. Conclusion:It was observed that HMGB1 detected in each group in Kidney specimens. the expression of HMGB1,CRE,BUN,TNF-α,NF-κB have the same trends in warm ischemia-reperfusion injury and cold ischemia-reperfusion injury, but cold ischemia-reperfusion injury have less elevated.It is considered that injury mechanism as the same as warm ischemia-reperfusion injury.Application of EP to block HMGB1 help to reduce the damage caused by renal cold ischemia-reperfusion injury.
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