应用白消安构建唯支持细胞综合征小鼠动物模型

Establishing a mouse model of Sertoli-cell-only syndrome by administration of busulfan

目的:构建稳定、可靠的小鼠唯支持细胞综合征模型。方法:60只SPF级NIH小鼠分成两组,分别给予30 mg/kg白消安腹腔注射及睾丸冷冻(0℃)30、60 min处理。于处理后2、4及8周,观察小鼠的生存率、睾丸重量及生精功能的变化,并对精子发生障碍程度进行定量化分析。结果:各组小鼠的基础体重值及处理后小鼠的生存率均无明显差异(P>0.05)。白消安处理后第4及8周,睾丸重量(g)(4周:0.04±0.01;8周:0.05±0.01)均显著低于对照组(4周:0.09±0.03;8周:0.11±0.02)(P<0.05),生精小管呈"唯支持细胞综合征"样改变,Johnsen评分(4周:3.86±0.50;8周:2.70±0.67)较对照组(4周:9.60±0.25;8周:9.76±0.43)均显著降低(P<0.01);睾丸冷冻30及60 min后,第2、4及8周睾丸重量(g)(冷冻30 min,2周:0.07±0.02;4周:0.06±0.01;8周:0.09±0.01)(冷冻60 min,2周:0.05±0.01;4周:0.04±0.02;8周:0.04±0.02)显著低于同期对照侧(2周:0.11±0.01;4周:0.11±0.01;8周:0.12±0.00)(P<0.05),睾丸形态学上易见生精小管萎缩、脱落、坏死及纤维化。各期冷冻组Johnsen评分(冷冻30 min,2周:4.70±0.67;4周;2.70±0.84;8周:6.10±1.14;冷冻60 min,2周:1.67±0.58;4周:1.20±0.45;8周:1.00±0.00)均显著低于同期对照组(2周:9.60±3.23;4周:9.60±0.55;8周:9.70±0.45)(P<0.01)。结论:白消安处理对小鼠的生存率无明显影响,可用来制备稳定、可靠的小鼠唯支持细胞综合征模型。

Objective： To establish a stable and reliable model of Sertoli-cell-only syndrome in mice. Methods ： We randomly divided 60 NIH mice into two groups of equal number to receive intraperitoneal injection of busulfan （30 mg/kg） and 30 or 60 minutes of testis cooling. At 2, 4 and 8 weeks after treatment, we reeorded the survival rate of the mice, weight of the testis and Johnsen scores and condueted quantitative analysis on the degrees of spermatogenetic failure. ResultS： There were no significant differences in the baseline body weight and survival rate between the intervention and control groups （ P 〉 0.05 ）. At 4 and 8 weeks, the testis weight and Johnsen score were significantly lower in the intervention group than in the control （ [0.04 ±0.01 ] g and [0.05 ±0.01 ] g vs [0.09 ±0.03] g and [0.11 ±0.02] g, P〈0.05; 3.86 ±0.50 and2.70 ±0.67 vs 9.60 ±0.25 and9.76±0.43, P〈0.01）. At 2, 4 and 8 weeks, the testis weights were （0.07 ± 0.02） g, （0.06 ± 0.01 ） g and （0.09 ± 0.01 ） g, respectively, in the 30-min cooling group and （0.05 ±0.01） g, （0.04 ±0.02） g and （0.04±0102） g in the 60-min cooling group, significantly lower than in the control side at the same time points （ [ 0.11 ±0.01 ] g, [ 0.11 ±0.01 ] g and [ 0.12± 0.00 ] g） （ P 〈 0.05 ）, and the Johnsen scores were 4.70 ± 0.67, 2.70 ± 0.84 and 6. 10± 1.14 in the 30-min and 1.67 ± 0.58, 1.20 ± 0.45 and 1.00 ± 0.00 in the 60-min cooling group, remarkably lower than in the control side （9.60 ± 3.23, 9.60 ± 0.55 and 9.70 ± 0.45 ） （P 〈 0.01 ）. Histopathological examination of the cooled testes revealed considerable atrophy of seminal tubules, necrosis of seminiferous epithelia and peritubular fibrosis. Conclusion ： Administration of busulfan has no obvious influence on the survival of mice, and is a reliable method for constructing a mouse model of Sertoli-cell-only syndrome.