【摘要】 背景:睾丸移植是短时间热缺血损伤后快速进入冷缺血的过程冷缺血时间的长短对睾丸生精功能是否存在影响目前还缺乏明确证据。目的:模拟临床睾丸移植缺血再灌注兔模型探索睾丸移植冷缺血时间与睾丸缺血再灌注损伤的相关性。设计、时间及地点:分组设计对比观察于2006-05/12在武汉大学人民医院动物实验中心完成。材料:雄性3～6月龄大白兔体质量2.5～3.5kg方法:采用自行设计的模拟临床睾丸移植过程的冷缺血灌注模型对20只大白兔左侧的睾丸采用0～4℃高渗枸橼酸腺嘌呤液恒压灌注并浸泡于0～4℃生理盐水中低温保存于冷缺血后1246h开放左侧睾丸血流。术后24h取双侧睾丸以右侧睾丸作自身对照。主要观察指标:苏木精-伊红染色及Johnsen评分判断睾丸损伤程度。丙二醛检测试剂盒检测每克睾丸组织中丙二醛含量。TUNEL法检测睾丸凋亡指数。结果:冷缺血1h后可见睾丸生精上皮结构稍紊乱2h后生精上皮开始脱落管腔内精子数量减少4h后生精上皮明显变薄仅见少量的精子细胞6h后生精上皮仅可见少量精母细胞部分呈唯Sertoli细胞表现。随冷缺血时间的延长睾丸缺血再灌注损伤逐渐加重Johnsen评分分值逐渐下降。睾丸组织冷缺血2h后再灌注24h的丙二醛水平明显高于自身对照组(P<0.05)至4h达到最高。凋亡细胞数量随冷缺血时间的延长逐渐增多凋亡指数高于自身对照(P<0.05)。结论:冷缺血后4h再灌注会造成睾丸生精上皮的严重损害提示睾丸移植前冷缺血时间应当控制在4h以内。更多还原

【Abstract】 BACKGROUND:Testes transplantation is the process of quick entrance of cold ischemia after the short time of warm ischemia injury.It still remains to determine whether the time of cold ischemia has effect on spermatogenic function.OBJECTIVE:To simulate the clinic ischemia-reperfusion model of testis transplantation,and to explore the relationship between the time of cold ischemia and ischemia-reperfusion injury of testes.DESIGN,TIME AND SETTING:This grouping,controlled observational experiment was performed in Animal Experiment Center of Renmin Hospital of Wuhan University from May to November 2006.MATERIALS:Male white rabbits of 3-6 months age,weighing 2.5-3.0 kg,were used in the experiment.METHODS:Twenty white rabbits were used to establish the clinic ischemia-reperfusion model of testes transplantation by self-designed method.The left testes were perfused with 0-4 ℃ hypertonic citrate adenine(HC-A) solution under constant pressure,and conserved in 0-4 ℃ normal saline.Reperfusions were performed at 1,2,4,and 6 hours after cold ischemia.Bilateral orchiectomies were performed at 24 hours after reperfusion.The right testes were assigned as self-controls.MAIN OUTCOME MEASURES:The degree of testicular injury was assessed by hematoxylin eosin staining and Johnsen score.The malondialdehyde(MDA) level per gram of testicular tissue was detected by MDA Elisa Kit.The apoptosis index(AI) of testes was determined by TUNEL.RESULTS:The structure disturbance of seminiferous epithelium was presented after one hour of the cold ischemia.Seminiferous cells began to shed and spermatozoa decreased after two hours of the cold ischemia.Seminiferous epithelium turned to thin obviously and little spermatid presented after four hours of the cold ischemia.Only a few spermatocytes presented,and some of them showed Sertoli cells after 6 hours of the cold ischemia.As the increasing cold ischemia time,the ischemia-reperfusion injury of testes aggravated and Johnsen score decreased.The MDA levels in injured testes at 24 hours after reperfusion were significantly higher than that in the self-controls(P < 0.05),and the peak appeared at 4 hours.The number of apoptotic cells in injured testes increased with the increase of cold ischemia time,and the AI was significantly higher than that in the self-controls(P < 0.05).CONCLUSION:The four-hour cold ischemia can result the severe seminiferous epithelium injury,which suggests testicular transplantation should be performed within 4 hours after cold ischemia.更多还原