维拉帕米及低温对大鼠睾丸扭转复位后生精功能的保护作用

Verapamil and Hypothermia Protect Spermatogenesis of Torsioned Testes in Rats

目的:探讨药物维拉帕米及低温对大鼠睾丸扭转复位后生精功能的影响。方法:60只健康青春期SD大鼠随机均分成5组,A组为睾丸扭转组,25℃室温,扭转时间2h;B组为睾丸扭转加维拉帕米组;C组为睾丸扭转加低温(9～13℃)组;D组为睾丸扭转加维拉帕米、低温组;E组为对照组。按Turner法制作睾丸扭转模型,取手术侧睾丸,光镜下观察组织学变化,行生精小管Makler评分,流式细胞术检测凋亡细胞百分率。结果:HE染色:除E组光镜下观察正常外,其余各组生精细胞排列不整齐,层次减少,生精细胞数量减少,可见凋亡小体及坏死区,部分区域可见炎症细胞浸润,其中扭转组最明显。生殖细胞凋亡率:A、B、C、D、E组分别为(32.11±2.20)%,(20.18±1.50)%,(20.02±1.90)%,(13.75±1.40)%,(8.56±0.90)%,与A组比较,B、C、D组凋亡率降低,差异有显著性(P<0.01)。Makler评分结果:A、B、C、D、E组分别为(14.47±1.35)、(15.45±0.75)、(15.48±0.75)、(16.22±0.72)、(19.60±0.56)分,与A组比较,B、C、D组Makler评分升高,差异有显著性(P<0.01)。结论:睾丸扭转复位后生精细胞凋亡增加可导致睾丸生殖能力下降;应用钙通道阻滞剂维拉帕米及局部低温均能增强睾丸组织的抗损伤能力;两者联合应用能更好地保护扭转复位睾丸的生殖功能。

Objective ： To investigate the protective effect of verapamil and hypothermia on the spermatogenesis of rats after testicular torsion. Methods ： Sixty healthy pubertal male Sprague-Dawley rats were equally divided into 5 groups ： A （ testis torsion） , B （ testis torsion ＋ verapamil）, C （testis torsion ＋ hypothermia）, D （testis torsion ＋ verapamil ＋ hypothermia）and E （control）. After treatment, the left testis was removed for the observation of the histological changes under the microscope and measurement of the percentage of apoptotic cells by flow cytometry. Results： HE staining showed disordered arrangement, reduced layers and decreased number of spermatogenic cells, apoptotic bodies, necrosis and partial invasion of inflammatory cells in all the groups but E, most obvious in Group A. The apoptosis rates of germ cells in Groups A, B, C, D and E were （32.11 ± 2.20）%, （20. 18± 1.50）%, （20.02 ± 1.90）%, （13.75 ± 1.40）% and （8.56 ± 0.90）%, respectively, and the Makler scores in the 5 groups were （14.47 ± 1.35）, （15.45 ± 0.75）, （15.48 ± 0.75）, （16.22 ± 0.72） and （19.60 ± 0.56）, respectively, both with statisti-cally significant differences （P 〈 0. 01 ）. Conclusion： The increased apoptosis of germ cells after testicular torsion-and-reposition may reduce the spermatogenesis of the testis. Either verapamil or local hypothermia can enhance testicular resistance against injuries, and the combination of the two can more efficiently prevent the germ eells from apoptosis.