| Objective(s):In this study,we established a rat model of renal ischemia-reperfusion injury(RIRI)to investigate the effects of recombinant human erythropoietih(rhEPO)and melatonin(Mel)on rat kidneys.The role of ischemia-reperfusion injury and preliminary study of its possible mechanism.Methods:A total of 32 healthy Sprague-dawley rats were randomly divided into 4 groups of 8 in each group:(1)Sham group(sham group):After anesthesia,the abdomen midline was used as an incision and the layers were opened layer by layer.At the abdomen,part of the intestine was removed from the body,the bilateral kidneys were detached,left and right renal pedicles were separated,and no ligation was performed.The incision was sutured 45 minutes after the incision was exposed;(2)Ischemia-reperfusion group(IR group):abdominal anesthesia The midline is an incision,laparotomy,part of the intestine was removed from the body,both sides of the kidney were isolated,left and right renal pedicles were separated,and bilateral renal pedicles were clipped with a non-invasive arterial clamp for 45 minutes to cause renal ischemia(renal changes from bright red Dark red),after 45 minutes,release the non-invasive arterial clip and perform reperfusion.Release the arterial clip and visually observe the color change of the kidney within 5 minutes.If the color of the kidney is changed from dark red to bright red after perfusion of the kidney,the perfusion is successful,and then the layers are sutured.Incision;(3)Erythropoietin group(EPO group):Recombineant human erythropoietih(rhEPO)(3000u/kg)was intraperitoneally injected pretreated 24 hours before surgery and established 24 hours after injection.mouse Ischemia and reperfusion model,surgical method referring to IR group;(4)melatonin group(Mel group):intraperitoneal injection of melatonin(Mel)(20mg/kg)was pretreated 24 hours before surgery.The rat model of ischemia-reperfusion was established 24 hours later.The surgical method was referred to IR group.The sham-operated group and the ischemic reperfusion group were given an equal volume of saline(0.7 ml/kg)intraperitoneally 24 hours before surgery.The rats in each group were sacrificed 24 hours after the blood was returned to the kidneys,and serum and kidney tissue specimens were also taken.Serum creatinine values(Scr)and blood urea nitrogen(BUN)were measured in each group;Hematoxylin-eosin staining(HE)staining was performed on each group of kidney specimens.Pathological changes;heme oxygenase-1(HO-1)was measured by immunohistochemistry to observe its expression in kidney tissue.Results:Analysis of renal function results:Compared with Sham group,serum creatinine(Scr)and urea nitrogen(BUN)were significantly increased in IR group,and the difference was statistically significant(P<0.01).Compared with IR group,creatinine(Scr)and urea nitrogen(BUN)were significantly decreased in EPO group and Mel group,and the difference was statistically significant(P<0.05).Compared with EPO group,serum creatinine(Scr)and urea nitrogen(BUN)in Mel group had no significant changes,and the difference was not statistically significant(P>0.05).Renal histomorphological analysis:The renal tissue structure of the Sham group was basically complete,with no obvious abnormalities.Compared with the kidney tissue of Sham rats,there were obvious morphological changes in renal tissue in IR group.It was obvious that renal tubular epithelial cells and glomerular capillary endothelial cells were significantly swollen,partially or even necrotic,a large number of caste were formed,and renal interstitial hyperemia was observed.Edema,a large number of inflammatory cell infiltration.Compared with the kidney tissue of the IR rats,the EPO group and the Mel group showed a small amount of inflammatory cell infiltration in the kidney tissue,and the degree of tissue damage was significantly reduced.Analysis of the expression of heme oxygenase-1(H0-1)in the kidney:In the Sham group,only a small amount of HO-1 expression was seen.Compared with the Sham group,the expression of HO-1 in the kidney was significantly increased in theIR group,and the difference was statistically significant.(P<0.05).Compared with IR group,the expression of HO-1 in kidneys of EPO group and Mel group was significantly increased,and the difference was statistically significant(P<0.05).Compared with the EPO group,there was no significant difference in HO-1 expression in the kidney of the Mel group(P>0.05).Conclusion(s):1.The rat kidney ischemia-reperfusion injury model was successfully prepared.Kidney ischemia-reperfusion injury can lead to the increase of serum creatinine and urea nitrogen,and cause renal damage.The expression of HO-1 in the kidney is increased.2.rhEPO pretreatment can significantly reduce the degree of renal damage,significantly reduce the pathological damage of the kidney.3.Mel pretreatment can significantly reduce the degree of renal damage,significantly reduce the pathological damage of the kidney.4.rhEPO and Mel can increase the expression of HO-1,we can speculate that rhEPO and Mel can reduce the oxidative stress response by inducing the expression of HO-1 in the kidney and play a protective role in renal IRI.5.rhEPO and Mel have similar levels of IRI protection in the kidney. |
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