| Objective:The rat renal ischemia-reperfusion(IR)model was established to explore the protective effect of edaravone on renal ischemia-reperfusion injury(IRI)and the protective mechanism of edaravone through mitochondrial signaling pathway.To provide some experimental evidence for clinical prevention and treatment of renal ischemia-reperfusion injury related diseases.Methods:Thirty healthy adult male Sprague-Dawley(SD)rats,SPF grade,weighing 220 to250 g,were randomly divided into three groups: Sham operation group(S group),Ischemia reperfusion group(IR group)and Edaravone group(E group).The surgical procedures included the establishment of renal IR model and caudal vein administration.The renal IR model was established by obtusly separating and clipping the bilateral renal artery for 45 min through the incision along the lower margin of the costal arch on both sides of the spine.This study is divided into three parts:1.The protective effect of edaravone on renal ischemia reperfusion injury in rats.Sham operation group: rats were only anesthetized,made incisions and isolated kidney treatment,no bilateral renal artery was clamped;Ischemia-reperfusion group,after bilateral renal artery were clipped for 45 min,the non-invasive arterial clip was released to restore blood supply;the Edaravone group: rats were given edaravone injection 3mg/kg 5 min before reperfusion via caudal vein.24 hours later,fresh blood and renal tissue samples were collected.The contents of creatinine and urea nitrogen in serum of rats in each group were determined,and the morphological changes of renal tissue were observed by HE staining with microscope.2.The effect of edaravone on mitochondrial apoptosis pathway during renal ischemia-reperfusion in rats.The apoptosis rate of renal tissue was determined by TUNEL staining.The expression of mitochondria-related apoptosis proteins Bcl-2,Bax and caspase-3 were detected by Western Bloting.3.Protective effect of edaravone on mitochondria during renal ischemia reperfusion and its mechanism in rats.The mitochondria of fresh kidney were isolated and purified,the changes of mitochondrial membrane potential of renal tissue were measured by JC-1,and the morphological changes of renal mitochondria were observed by transmission electron microscope.Results:1.Edaravone preconditioning alleviated renal ischemia reperfusion injury.Renal ischemia reperfusion model was successfully constructed in rats.Compared with group S,serum creatinine(Cr)and blood urine nitrogen(BUN)were significantly increased(P<0.05),renal tubular injury semi-quantitative score were significantly increased of the rats in IR group(P<0.05).Compared with IR group,serum creatinine(Cr)and blood urine nitrogen(BUN)levels were significantly decreased(P<0.05);renal tubular injury semi-quantitative score were evidently decreased in the E group(P<0.05).2.The apoptosis rate and caspase-3 expression in renal tissue were significantly increased in ischemia-reperfusion group(P<0.05),but the apoptosis rate and caspase-3were significantly decreased after edaravone treatment(P<0.05).Ischemia-reperfusion increased the expression of Bax and decreased the expression of Bcl-2 pronouncedly,leading to a significantly reduction of the radio of Bcl-2/Bax(P<0.05),compared with those of sham operation group.Edaravone pretreatment significantly decreased theexpression of Bax and increased the expression of Bcl-2(P<0.05),giving rise to significantly rise of the ratio of Bcl-2/Bax(P<0.05).3.Compared with the sham-operated group,the mitochondrial membrane potential was significantly decreased(P<0.05),the mitochondrial injury was aggravated and the pathological injury score was significantly increased(P<0.05).The mitochondria membrane potential in renal tissue was significantly increased(P<0.05),the mitochondrial damage was alleviated and the pathological injury score was significantly decreased in rats pretreated with edaravone(P<0.05).Conclusion:1.Edaravone preconditioning can attenuate the injury of renal structure and function caused by ischemia reperfusion injury in rats.2.Edaravone reduced renal ischemia-reperfusion injury in rats by inhibiting mitochondrial apoptosis pathway and reducing apoptosis rate of renal tissue cells in rats.3.The protective effect of edaravone on renal ischemia-reperfusion injury is related to the maintenance of mitochondrial structure(reducing the damage of mitochondria ultrastructure)and function(maintaining the mitochondrial membrane potential). |
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