Study On The Mechanism Of Rabbit Testicular Ischemia Reperfusion Injury And Its Protective Measures

ObjectiveTo establish rabbit testicular ischemia reperfusion injury model based on testicular torsion, and then combined with oxidative stress indexs and cell apoptosis factors examination to explore the protection of ischemic preconditoning, postconditioning and combined treatment on it and the potential mechanism.MethodsA total of 30 New Zealand white rabbits (male) randomly divided into 5 groups, including A group (control group), exposing right spermatic cord and no subjecting to ischemia; B group (ischemia reperfusion group), occlusion right spermatic cord for 60 min using noninvasive clamp and following with 3 days reperfusion; C group (ischemic preconditoning group), occlusion spermatic cord 3 times (ischemic 5 min/time+ reperfusion 5 min/time) before ischemic reperfusion, and occlusion right spermatic cord for 60 min using noninvasive clamp and following with 3 days reperfusion; D group (ischemic postconditioning group), after occlusion right spermatic cord for 60 min, occlusion spermatic cord 3 times (ischemic 5 s/time+reperfusion 5s/time) and following with 3 days reperfusion; E group (Combined treatment), occlusion spermatic cord 3 times (ischemic 5 min/time+reperfusion 5 min/time) before ischemic reperfusion, and occlusion right spermatic cord for 60 min using noninvasive clamp and subjecting to occlusion spermatic cord 3 times (ischemic 5s/time+reperfusion 5s/ time) and following with 3 days reperfusion. Subsequently, rabbits were narcotized with 3% barbital sodium with collecting of the whole blood to separate serum, and then examined the content of testosteone. Then, testicular tissues were seperated and grouped to torsion side and uninjuried side, and the MDA, PC, NO, SOD, MPO and GSH-Px expression level was examined by ELISA kits after splitting, the Bcl-2, Bax expression level was analysed by Western blot assay and another tissues were fixed using 10% neutral formalin and performed to HE staining and TUNEL staining.ResultsIn contrast to normal group A, the testosteone expression was significantly decreased in torsion group B, ischemic preconditoning group C, ischemic postconditioning group D and combined treatment group E (p<0.01), and the testosteone expression was also obviously decreased in torsion group B, ischemic preconditoning group C, ischemic postconditioning group D and combined treatment group E in contrast of preoperation and postoperation (p<0.05). In comparison of normal group A and uninjuried side of torsion group B, the expression of MDA, PC, NO, SOD, MPO and GSH-Px was significantly increased in torsion side of torsion group B, and was significantly decreased in ischemic preconditoning group C, ischemic postconditioning group D and combined treatment group E in contrast of preoperation and postoperation (p<0.01), and had no statitical difference of these three treatments. Pathological staining results exhibition, in comparison of normal group A and uninjuried side of torsion group B, the major of seminiferous tubule damage, spermatogenic cells disappearing, some apoptosis spermatogenic cells, light eosion edema fluid exudation in mesenchyme and lumen, apoptosis index obviously incresaing (p<0.01) and Johnsen grade significantly decreasing (p<0.01) in torsion side of torsion group B, and restored to normal after ischemic preconditoning, postconditioning and combined treatment. Similarly, the Bcl-2/Bax ratio was significantly decreased in torsion side of torsion group B (p<0.01) when compared to that of normal group A and uninjuried side of torsion group B, and restored to normal after ischemic preconditoning, postconditioning and combined treatment.ConclusionIschemic preconditoning, postconditioning and combined treatment has an obvious improvement on rabbit testicular ischemia reperfusion injury through regulating oxidative stress index and cell apoptosis, and these three treatment had same protection. This study not only provided a significant technical reference for studying the potential mechanism of ischemia reperfusion, and also provided a potential drugs for clinical treatment of ischemia reperfusion injury, and exhibited the momentous value of clinical application.