论文部分内容阅读
目的:基于睾丸扭转建立兔睾丸缺血再灌注损伤模型,探索缺血预处理对睾丸缺血再灌注损伤兔的睾酮水平及生精细胞凋亡的影响。方法:雄性新西兰大白兔15只,随机分为3组:A组(对照组)暴露右侧精索,不作缺血处理并关闭;B组(缺血再灌注组)使用无创血管夹夹闭右侧精索致睾丸缺血60 min,再灌注3 d;C组(预处理组)预先夹闭精索3次(缺血5 min/次+灌注5 min/次),余同B组。造模完成后,采用3%巴比妥钠麻醉兔后采集血液并分离血清,测定血清睾酮含量;随后分离睾丸组织,按照缺血侧及健侧分组后采用10%中性福尔马林固定组织,进行HE染色和TUNEL细胞凋亡检测。结果:术后A、C组体重比术前明显增加[(2.65±0.07)kg vs(2.45±0.07)kg、(3.03±0.11)kg vs(2.92±0.07)kg](P均<0.05),B组手术前后体重没有变化[(3.05±0.07)kg vs(3.05±0.07)kg](P>0.05)。睾酮含量:手术前后A组没有明显差异[(139.59±9.39)ng/L vs(140.19±9.47)ng/L,P>0.05];术后B、C组显著低于术前[(74.12±4.00)ng/L vs(148.06±3.31)ng/L、(94.76±3.13)ng/L vs(133.75±6.48)ng/L]和术后A组(P均<0.01)。病理结果显示,与A组及B组健侧相比,B组缺血侧睾丸组织内大部分生精小管结构破坏,生精小管内各级生精细胞结构消失,可见凋亡的生精细胞,间质及管腔内有淡伊红色水肿液渗出,凋亡指数明显升高(P<0.01),Johnsen评分显著降低(P<0.01);与B组缺血侧相比,C组缺血侧睾丸组织基本恢复正常,凋亡指数明显降低(P<0.01),Johnsen评分显著增加(P<0.01)。结论:缺血预处理明显缓减了睾丸缺血再灌注损伤后睾酮水平下降,并降低了细胞凋亡,为睾丸缺血再灌注损伤的临床治疗提供了潜在干预措施。
OBJECTIVE: To establish a model of testicular ischemia-reperfusion injury in rabbits based on testicular torsion and to explore the effect of ischemic preconditioning on testosterone and spermatogenic cell apoptosis in testicular ischemia-reperfusion injury rabbits. Methods: Fifteen male New Zealand white rabbits were randomly divided into three groups: group A (control group) exposed the spermatic cord on the right without ischemia and closed; Group B (ischemia-reperfusion group) Lateral spermatic cord induced testicular ischemia 60 min, reperfusion 3 d; C group (pretreatment group) pre-clamp spermatic cord 3 times (ischemia 5 min / times + perfusion 5 min / times), the same with the B group. After modeling, the rabbits were anesthetized with 3% sodium pentobarbital to collect blood and serum was collected to measure the serum testosterone content; then isolated testicular tissue, according to the ischemic and contralateral grouping with 10% neutral formalin fixation Tissues were HE staining and TUNEL apoptosis assay. Results: The body weight of patients in group A and group C were significantly higher than that before operation [(2.65 ± 0.07) kg vs (2.45 ± 0.07) kg vs (3.03 ± 0.11) kg vs (2.92 ± 0.07) kg respectively (P <0.05) There was no change in body weight before and after operation in group B [(3.05 ± 0.07) kg vs (3.05 ± 0.07) kg] (P> 0.05). The level of testosterone in group B and C was significantly lower than that in group B before and after surgery [(139.59 ± 9.39) ng / L vs (140.19 ± 9.47) ng / L, P> 0.05] (148.06 ± 3.31) ng / L, (94.76 ± 3.13) ng / L vs (133.75 ± 6.48) ng / L] and postoperative A group (all P <0.01). Pathological results showed that most of the seminiferous tubules in testis of group B were damaged compared with that of group A and B, and the structure of spermatogenic cells disappeared at all levels in seminiferous tubules. The number of apoptotic spermatogenic cells (P <0.01), and Johnsen score decreased significantly (P <0.01). Compared with the ischemia group B, the group C lacked Blood testis tissue returned to normal, apoptosis index was significantly lower (P <0.01), Johnsen score was significantly increased (P <0.01). CONCLUSION: Ischemic preconditioning can significantly reduce testosterone level and decrease apoptosis after testicular ischemia-reperfusion injury, providing a potential intervention for the clinical treatment of testicular ischemia-reperfusion injury.