白消安注射法建立昆明小鼠精原干细胞移植受体模型

Establishment of transplantation model for spermatogonial stem cells in Kunming mice with busulfan treatment

【目的】 通过对6周龄昆明小鼠腹腔内单次注射不同剂量的白消安来制作精原干细胞移植受体模型。【方法】将104只昆明小鼠随机分成4组，分别注射0（对照组，注射等量的白消安溶解液二甲基亚砜（DMSO），10，20和30 mg/kg白消安，于注射白消安后5，7，9，11和13周记录小鼠的死亡数、小鼠睾丸外观体积、小鼠睾丸质量和小鼠体质量，计算睾丸指数；通过组织学观察注射不同剂量白消安后小鼠曲细精管精子恢复情况，并检测受体小鼠血液中红细胞和白细胞数量的变化。【结果】 注射0（对照组），10，20和30 mg/kg白消安13周后，小鼠死亡率分别为0（0/22），11.54%（3/26），17.86%（5/28）和89.29%（25/28）。10 mg/kg白消安组小鼠的睾丸指数在注射后5，7和9周显著小于对照组（P＜0.05），而在注射后11和13周，与对照组差异不显著（P＞0.05）；在注射后不同时间，对照组和10和20 mg/kg白消安组的睾丸指数均显著高于20 mg/kg白消安组（P＜0.05）。在注射白消安5，7，9，11和13周后，对照组曲细精管上皮无变化；10和20 mg/kg白消安组有完整精子发生的曲细精管占总曲细精管的百分比分别为（49.56±8.23）%，（66.61±6.13）%，（80.24±10.42）%，（93.87±3.23）%，（92.49±4.58）%；（8.73±9.22）%，（9.89±6.39）%，（29.21±12.31）%，（49.64±3.59）% 和（81.99±4.98）%。受体小鼠红细胞和白细胞数量均随着处理时间及白消安处理剂量的变化而变化。【结论】腹腔内注射20 mg/kg白消安小鼠的死亡率、睾丸指数均较低，中空的曲细精管数量多，且恢复较慢，适于做精原干细胞移植的受体，合适的移植时间是白消安处理后5～7周。

[Objective] Different doses of busulfan were injected to Kunming （KM） mice at age of 6 weeks by intra-abdominal to produce receptor model for transplantation of spermatogonial stem cells. [Method] A total of 104 male KM mice were randomly divided into 4 groups and treated with different doses of busulfan （0,10,20 and 30 mg/kg body weight）. weight, testicle weight, the testicular index, the number After 5,7,9,11 and 13 weeks, mortality, body of seminiferous tubule with spermatogenesis and blood cells were analyzed. [Result] The mortalities were 0 （0/22）, 11. 54% （3/26）, 17.86% （5/28） and 89.29 % （25/28） for 0,10,20 and 30 mg/kg groups, respectively. Testieular indexes in control were significantly higher than in 10 mg/kg group 5,7,and 9 weeks after busulfan treatment （P〈0.05）. However, 11 and 13 weeks after busulfan treatment,they were insignificantly different （P〉0.05）. Regardless of injecting dose,testicular index in 20 mg/kg group were significantly higher than the control and 10 mg/kg group （P〈0. 05） all the time. Seminiferous epithelia of the control group had no obvious degenerative changes after 5,7,9,11 and 13 weeks. The tubules with complete spermatogenesis were （49.56±8.23）%,（66.61±6.13） %, （80.24±10.42） %, （93.87±3.23）% and （92.49±4.58） % in 10 mg/kg group. （8.73± 9.22）%,（9.89±6.39）%,（29.21±12.31）% ,（49.64±3.59）% and （81. 99±4.98）% of seminiferous tu-bules showed complete spermatogenesis in 20 mg/kg group after 5,7,9,11 and 13 weeks. The number of blood cells changed along with time and dose of busulfan treatment. [Conclusion] The optimal dose of busulfan was 20 mg/kg and the most appropriate time for transplantation was 5--7 weeks after busulfan treatment.