| Purpose:build the model of rat cervical kidney transplantation to discuss the protective function and relevant mechanism of rat kidney transplantation with continuous hypothermical perfusion of alprostadil to rat renal transplantation's warm ischemia,so as to provide experimental evidence for alprostadil's application to the clinic organ preservation.Methods:Select inbred line SD rats as the experimental target,they are male rats with average weight of about 250-300 g.Employ completely random design(random number table)to group 80 donor SD rats into two groups-control group(A group)and experimental group(B group),each group will accept detection at four timings-one day,three days,five days,seven days.The rats' kidneys of two groups are all in soaking ischemia for 45 min,then are perfused 2h continuously under 4?condition.After perfusion,make a cervical kidney transplantation of donor kidney and extirpate two kidneys of the acceptor.Control group: the perfusate is HC-A kidney preserving fluid+normal saline;experimental group: the perfusate is HC-A kidney preserving fluid + Lipo-PGE1.Raised in the clean environment after operation,the rats are reperfused into blood after one day,three days,five days and seven days,collect jugular blood,detect blood urea nitrogen,serum creatmne content by full-automatic biochemistry analysis instrument,put them to death after blood sampling and extract kidney tissue.Make use of enzyme-linked immunosorbent assay to detect the kidney tissue's 8-hydroxy—2deoxyguanosine,malondialdehyde content,detect caspase-3 and hypoxia inducible factor-lalpha's expression through western blotting;after HE dying of kidney tissue,observe pathomorphology.Utilize SPSS18.0software to dispose and analyze the received data.Result:1.Renal function the serum BUN concentration of two groups' rat all appears the rising trend,and conduct an intra-class comparison of two groups 1day,three days,five days after operation,the difference has a statistical meaning(P<0.05),there is no apparent difference in the comparison between five days' and seven days' after operation(P>0.05);compare all intra-class timings after operation,group B is obviously lower than group A(P<0.05).the rat serum Cr concentration of two groups appears the rising trend as time prolongs(P<0.05),which will reach to the peak 7 days after operation;compare the same intra-class timing of two groups,group B is apparently lower than group A(P<0.05).2.Renal histomorphology change the histomorphology change is apparent in the different timings of light microscope observation in group A,B.The glomerulus enlarges in group A,glomerular capillary has a vague outline,renal capsule space becomes small,many renal tubular epithelium has edema-degeneration or necrosis,intracytoplasm red-dying particle number is increasing apparently and thickening,couples of small focus tubular necrosis can be seen.In group B,glomerulus enlarges,glomerular capillary has a clear outlook,the decrease of renal capsule space is no apparent,small renal tubular epithelium has edema-degeneration or necrosis,intracytoplasm red-dying particle number is increasing a little but not thickening,small quantity of rsd cell casts can be in bureaucratic tone,there is no breaking,necrosis scene in renaltubular epithelial cell.3.Renal tissue 8-OHDG and MDA content the 8-OHDG content of two groups appear the rising trend as time prolongs after reperfusion;compare the same intra-class timing of two groups after operation,the8-OHDG content of all bordering timings in group B has no apparent change(P>0.05);while the 8-OHDG content variation is apparent in the first day,the third day,the fifth day after operation in group A(P<0.05).conduct an intra-class comparison in the first day,the third day,the fifth day,the seventh day after operation,the 8-OHDG content in group B has an apparent decrease than group A(P<0.05).The MDA content of two groups appears a rising trend in the first day,the third day,the fifth day after operation,and reaches to the peak in the fifth day after operation,therewith is decreasing in varying degrees.The MDA content in group B is apparently lower than group A in the first day,the third day,the fifth day,the seventh day after operation(P<0.05);compare in group,the MDA content has an apparent decrease in the seventh day after operation than the fifth day's in group B(P<0.05),the MDA content has no apparent difference in the seventh day after operation than the fifth day's in group A(P>0.05).4.Renal tissue caspase-3 and HIF-1? protein's relative transcript level compare in the first day,the third day,the fifth day,the seventh day after operation in two groups,caspase-3 expression level in group B is apparently lower than group A(P<0.05);caspase-3 expression level appears a rising trend in group A,and reaches to the peak in the fifth day after operation,then go down slowly in the seventh day,the intra-class comparison in the first day,the fifth day,the seventh day after operation has an apparent difference(P<0.05),while the comparison in the fifth day,the seventh day are not apparent(P > 0.05).The caspase-3 expressionlevel in group B reaches to the peak value in the third day after operation,the decreases apparently in the fifth day,the seventh day(P<0.05).the HIF-1? expression level in group B is apparently lower than group A(P<0.05).Conclusion:1.the HC-A renal preserving fluid with Lipo-PGE1 enables to relief lipid peroxidation damage and DNA oxidative damage induced by oxygen radical effectively,to alleviate damage and apoptosis of renal tubular epithelial cell,so as to improve preserving quality of warm-ischemia kidney,and enhance the renal function of transplanted rat.2.Oxidative stress and renal tubular epithelial cell apoptosis play an important role in the rat renal transplantation ischemia reperfusion injury mechanism. |
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