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目的:探讨单次热应激对小鼠睾丸生精功能可复性研究。方法:C57雄性小鼠36只,随机按1、7、14 d各分为对照组(n=6)和热应激组(n=6)。热应激组和对照组小鼠下腹部分别在43℃和25℃水浴中干预15 min,于处理后1、7、14 d取材。计算睾丸器官指数,HE染色观察睾丸组织结构变化,免疫组化染色检测早幼粒白血病锌指蛋白(PLZF)和联会复合体蛋白3(SCP-3)在睾丸组织中的定位及表达,Western印迹检测PLZF蛋白表达水平。结果:热应激组小鼠睾丸器官指数明显低于对照组(P<0.01)。HE染色可见,热应激组与对照组相比,热应激后第1天生精细胞排列松散,散落在生精小管中;第7天生精小管萎缩,排列疏松,生精细胞明显减少;第14天生精小管排列较为紧密,生精细胞数量增加,生精细胞层数增多。与对照组相比第1、7天热应激组SCP-3标记的精母细胞明显减少,第14天精母细胞数量开始恢复。Western印迹检测结果显示,PLZF蛋白在热应激处理后1 d表达量显著降低(0.19±0.02 vs 0.64±0.03,P<0.01),第7、14天又迅速增加(0.77±0.02、0.77±0.02),且显著高于对照组(P<0.01)。结论:小鼠睾丸热应激处理后可致生精功能发生障碍,随着时间的推移,这种损伤具有可复性。
AIM: To investigate the refolding effect of single heat stress on spermatogenesis in mice testes. Methods: Thirty - six C57 male mice were randomly divided into control group (n = 6) and heat stress group (n = 6) on day 1,7 and 14. The lower abdomen of heat stress group and control group mice were respectively treated in water bath at 43 ℃ and 25 ℃ for 15 min, and were drawn at 1, 7 and 14 days after treatment. The testis organ index was calculated and the histological changes of testis were observed by HE staining. The localization and expression of PLZF and SCP-3 in testis tissue were detected by immunohistochemistry. Blot detection of PLZF protein expression levels. Results: The organ index of testis in heat stress group was significantly lower than that in control group (P <0.01). Compared with the control group, the number of spermatogenic cells in the heat-stressed group was loosely arranged and scattered in the seminiferous tubules on the first day after heat stress. On the 7th day, the seminiferous tubules were atrophied and loosely arranged, and the spermatogenic cells were significantly reduced. 14 day-old seminiferous tubules arranged more closely, the number of spermatogenic cells increased, the number of spermatogenic cells increased. Compared with the control group, the number of spermatids marked by SCP-3 in the heat stress group decreased significantly on the first day and the number of spermatocytes began to recover on the 14th day. The results of Western blotting showed that the expression of PLZF protein decreased significantly on the 1st day after heat stress (0.19 ± 0.02 vs 0.64 ± 0.03, P <0.01), and rapidly increased on the 7th and 14th day (0.77 ± 0.02, 0.77 ± 0.02 ), And significantly higher than the control group (P <0.01). CONCLUSIONS: Treatment of heat stress in mice testes impaired function of spermatogenesis, with recalcitrance over time.