Objective To explore the apoptosis and the express of proliferating cell nuclear antigen (PCNA)in spermategenie cells,and study generation function of the rabbit after serotal reconstruction with flaps.Methods The 48 New Zealand white rabbits w.ere randomly divided into three groups as experimental group(the scrotum reconstructed with flaps,n=48),the control group(the sham operated group,n=24)and the blank group(n=18).The apoptosis and the expression of PCNA in the spermatogenie cells were detected with TUNEL and the immunohistochemistry from the 3rd to the 8th week after operation.8 weeks later,12 animals in each group were fed respectively with one female rabbit to observe the procreation.Results The apeptotie index of the spermatogenic cells in blank group waB 7.73±4.95.3 weeks after operation.the apeptotie index of spermatogenic cells wag 22.59±3.04 in the experimental group,and 21.13±1.68 in control group,showing no significant difference between the two groups(P＞0.05).8 weeks after operation,the apeptotie index of spermatogenic cells was 71.85±2.69 in the experimental group,and 13.64±2.09 in control group,show a significant difference between them (P＜0.05).The apoptotic index in experimental group increased gradually from the 3rd to 8th week after scrotal reconslruction ,which was markedly higher than that in the blank group(P＜0.05).The apaptefic index in control group was higher than that in the blank group at the 3rd week(P＜0.05),but not at the 8th week (P＞0.05).The proliferation index of spermatogenic cells was 9.32±9.30 and 12.52±3.87 in experimental group at the 3rd and 4th week,respectively,which was significantly lower than that in blank group(43.07±2.25) and control group(45.69±4.98) at the 3rd week(P ＜ 0.05).The proliferation index of spermatogcnic cells was 46.98±18.92 and 49.53±9.79 in experimental group at the 7th and 8th week,respectively,39.90±5.10 in control group at the 8th week,showing no difference between the two groups (P＞0.05).The proliferation index of spermatogenic cells in control group at the 3rd and 8th week was not different from that in the blank group(P＞0.05)The female pairing rabbits in the blank and control group were all pregnant,and the average childbirths were 6.0±1.28 and 5.92±1.31 respectively,with no difference between the two groups(P＞0.05).All the female pairing rabbits in the experimental group were not pregnant,showing a significant difference from those in the blank and control group(P＜0.05).Conclusions The rabbit generation functional disturbance after scrota1 reconstruction with flaps is due to the excessive apeptosis of spermatogenic cell.The spermatogcnic cell proliferation is affected only in the early postoperative period,but can recover later.%目的 通过检测皮瓣重建阴囊后睾丸生精细胞凋亡及增殖细胞核抗原(PCNA)表达情况,以探讨皮瓣重建阴囊后对兔生殖功能的影响.方法 以育龄期新西兰大白兔作为实验动物,雄性动物随机分为实验组(皮瓣重建阴囊组)48只、对照组(皮瓣重建阴囊假手术组)24只和空白组18只.建模后的第3～8周末应用原位末端标记法(TUNEL)检测生精细胞凋亡和免疫组织化学法(SABC法)检测PCNA表达情况;第8周末各组分别对未取睾丸活检的12只雄性家兔与雌兔配对喂养,观察生育情况.结果 凋亡检测结果 :空白组正常睾丸生精细胞的凋亡指数为7.73±4.95.在第3周末实验组凋亡指数为22.59 4-3.04,对照组为21.13±1.68;在第8周末实验组凋亡指数为71.85±2.69.对照组为13.64±2.09.实验组在重建阴囊后3～8周内随着时间延长生精细胞凋亡指数逐渐增加,其凋亡指数明显高于空白组(P＜0.05);在第3周末实验组与对照组凋亡指数比较差异无统计学意义(P＞0.05);在第8周末实验组与对照组凋亡指数比较差异具有统计学意义(P＜0.05);第3周末对照组凋亡指数与空白组比较差异具有统计学意义(P＜0.05);第8周末对照组凋亡指数与空白组比较差异无统计学意义(P＞0.05).PCNA检测即增殖指数检测结果 :空白组正常睾丸生精细胞的增殖指数为43.07±2.25.第3周末及第4周末实验组增殖指数分别为9.32±9.30及12.52±3.87,对照组第3周末为45.69±4.98,实验组第3、4周末增殖指数分别与对照组第3周末及空白组增殖指数比较差异具有统计学意义(P＜0.05);实验组第7周末及第8周末增殖指数分别为46.98±18.92及49.53±9.79,对照组在第8周末为39.90±5.10,实验组第7、8周末增殖指数分别与对照组第8周末及空白组增殖指数比较差异无统计学意义(P＞0.05);对照组增殖指数在第3周末和第8周末分别与空白组增殖指数比较差异均无统计学意义(P＞0.05).配对结果 :空白组、对照组配对后对应母兔均受孕生崽,平均生崽数分别为(6.0±1.28)只、(5.92±1.31)只,两组生崽数差异无统计学意义(P＞0.05);实验组配对后对应母兔均未受孕,与空白组和对照组比较差异有统计学意义(P＜0.05).结论 皮瓣重建阴囊导敛的家兔生殖功能障碍主要是由于生精细胞的过度凋亡所致,生精细胞的增殖能力仪在重建阴囊早期受到一定的影响.而后期则无明显受损表现.
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