摘要
目的:量化评价白消安二次腹腔注射制备的小鼠精子再生模型。方法:54只8～10周龄雄性昆明白小鼠随机分为对照组和两个模型组,每组18只。模型组小鼠分别以10mg/kg和15mg/kg白消安间隔24d二次腹腔注射建立精子再生模型,对照组小鼠以50%二甲基亚砜溶液10ml/kg间隔24d二次腹腔注射。采用Johns-en评分量化给药后3、4和8周时生精上皮精子发生,运用实时定量PCR技术检测这3个时期支持细胞GATA结合蛋白4(GATA-4)mRNA和胶质细胞源性神经营养因子(GDNF)mRNA表达水平。结果:对照组Johnsen评分在各时期无显著差异(P>0.05)。给药后3周和4周,两模型组Johnsen评分均显著低于对照组(P<0.01);给药后4周和8周,15mg/kg组Johnsen评分均低于10mg/kg组(P<0.05);给药后8周,15mg/kg组Johnsen评分仍显著低于对照组(P<0.05),而10mg/kg组和对照组间无显著差异(P>0.05)。各组各时期支持细胞GATA-4mRNA表达均无显著差异(P>0.05)。对照组各时期支持细胞GDNF mRNA表达无显著差异(P>0.05)。两模型组支持细胞GDNF mRNA表达在给药后3周均高于对照组,而在给药后4周均低于对照组(P<0.01);给药后8周,15mg/kg组支持细胞GDNF mRNA表达低于对照组(P<0.01),而10mg/kg组与对照组无显著差异(P>0.05)。在以上3个时期,15mg/kg组GDNF mRNA表达均低于10mg/kg组(P<0.01)。结论:10mg/kg白消安间隔24d二次腹腔注射是建立小鼠精子再生模型的适宜剂量,增大剂量可使支持细胞GDNF mRNA表达不足,导致精子发生不能完全恢复。
Objective：To quantitatively evaluate the murine model of spermatogenesis regeneration induced by two-dose busulfan injection.Methods：Fifty-four male mice were randomly divided into a control and two model groups of equal number,the former treated by two-dose intraperitoneal injection of 50% DMSO solution at 10 ml/kg,and the latter by that of busulfan at 10 mg/kg and 15 mg/kg respectively to establish spermatogenesis regeneration models,both at the interval of 24 days between the two doses.Spermatogenesis in seminiferous epithelia was evaluated by Johnsen score,and the expressions of GATA-4 and GDNF mRNA in Sertoli cells were detected by real time quantitative PCR at 3,4 and 8 weeks after the treatment.Results：Johnsen score kept stable in the control group at all stages （P0.05）,but higher than in the model groups at 3 and 4 weeks （P0.01）.It was lower in the 15 mg/kg than in the 10 mg/kg model group at 4 and 8 weeks （P0.01）,and than in the control group at 8 weeks （P0.05）,but had no significant difference between the 10 mg/kg and the control groups （P0.05）.Nor did the expression of GATA-4 mRNA in Sertoli cells show any significant difference among the three groups at different stages after the treatment （P0.05）,and that of GDNF mRNA at different stages in the control group （P0.05）.Compared with the controls,the level of GDNF mRNA in Sertoli cells was significantly higher at 3 weeks but lower at 4 weeks in the model groups （P0.01）,and lower in the 15 mg/kg group （P0.01） and comparable in the 10 mg/kg group at 8 weeks （P0.05）;and it was lower in the 15 mg/kg than in the 10 mg/kg group at all stages （P0.01）.Conclusion：Two-dose intraperitoneal injection of 10 mg/kg busulfan at the interval of 24 days is an optimal option for the establishment of a murine model of spermatogenesis regeneration.Higher dose of busulfan may induce deficient expression of GDNF in Sertoli cells and result in incomplete restoration of spermatogensis.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2010年第5期395-399,共5页
National Journal of Andrology
基金
国家自然科学基金(30400160)~~
关键词
白消安
精子再生
量化评价
支持细胞
小鼠
busulfan
spermatogenesis regeneration
quantitative evaluation
Sertoli cell
mouse
