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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 mouse sperm regeneration model prepared by Baixian’an intraperitoneal injection. Methods: Fifty-five male Kunming white mice aged 8-10 weeks were randomly divided into control group and two model groups, with 18 rats in each group. The mice in model group were injected intraperitoneally with 10 mg / kg and 15 mg / kg busulfan for 24 days, respectively. The mice in control group were injected intraperitoneally with 10 ml / kg 50% dimethyl sulfoxide for 24 days. The spermatogenic epithelial spermatogenesis was quantified by Johns-en score at 3, 4 and 8 weeks after the administration, and GATA-4 mRNA and glial cell-derived neurotrophic factor were detected by real-time quantitative PCR Factor (GDNF) mRNA expression level. Results: The Johnsen score of the control group had no significant difference at all stages (P> 0.05). The Johnsen scores of the two model groups were significantly lower than those of the control group at 3 weeks and 4 weeks after administration (P <0.01). Johnsen scores of the 15 mg / kg group were lower than those of the 10 mg / kg group at 4 and 8 weeks after administration <0.05). At the 8th week after administration, the Johnsen score of 15mg / kg group was still significantly lower than that of the control group (P <0.05), while there was no significant difference between the 10mg / kg group and the control group (P> 0.05). There was no significant difference in the expression of GATA-4mRNA among the three groups (P> 0.05). There was no significant difference in the expression of GDNF mRNA in the control group at each time point (P> 0.05). GDNF mRNA expression in the two model groups was significantly higher than that in the control group at 3 weeks after administration and at 4 weeks after administration (P <0.01). At 8 weeks after administration, GDNF mRNA expression in the 15 mg / kg group (P <0.01), while there was no significant difference between the 10 mg / kg group and the control group (P> 0.05). During the above three periods, the expression of GDNF mRNA in 15mg / kg group was lower than that in 10mg / kg group (P <0.01). Conclusion: The second intraperitoneal injection of busulfan at 10mg / kg for 24 days can reduce the expression of GDNF mRNA in supportive cells, resulting in incomplete recovery of spermatogenesis.