pH2AX蛋白在单次热应激致小鼠睾丸生殖功能可逆性中的表达变化  

Dynamic changes of pH2AX expression in the reversibility of mouse testicular reproductive function impaired by single heat stress

作　　者：田洪成 徐颖桥 马鹤[1] 王会君[1] 李学峰[1] 王新花 马良宏[3] 李清春[1] 张冬梅[1] 王雁林[1] 

机构地区：[1]滨州医学院附属医院生殖医学科,山东滨州256600 [2]威海市妇幼保健院生殖医学科,山东威海264200 [3]宁夏医科大学总医院泌尿外科,宁夏银川750004

出　　处：《中华男科学杂志》2017年第10期873-877,共5页National Journal of Andrology

基　　金：滨州医学院科技计划项目(BY2016KJ60)~~

摘　　要：目的:探讨pH2AX在单次热应激致小鼠睾丸生殖功能可逆性中的表达变化。方法:C57雄性小鼠24只,随机按1、7、14 d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置于43℃和25℃水浴中干预15 min,于处理后1、7、14 d取睾丸组织。TUNEL检测睾丸细胞凋亡情况,免疫组化染色检测pH2AX蛋白定位及表达,Western印迹检测pH2AX蛋白表达水平。结果:TUNEL结果显示,在热应激后的第1天生精小管中的凋亡细胞数量最多,第7、14天细胞凋亡基本消失。免疫组化显示,pH2AX蛋白在睾丸生精小管基底膜处细胞核表达。Western印迹检测显示,与对照组相比,热应激后的第1天,pH2AX蛋白表达明显增加(0.47±0.02 vs 1.61±0.04,P<0.01);第7天(0.85±0.03)与热应激处理第1天相比pH2AX蛋白表达显著减少(P<0.01);第14天(1.72±0.02)与热应激处理第1、7天及对照组相比pH2AX蛋白表达显著增加(P均<0.01)。结论:小鼠睾丸热应激处理后pH2AX蛋白表达量在睾丸中出现动态变化,这种变化与热应激后睾丸生精细胞的增殖分裂相关。Objective： To investigate the role of pH2AX in the reversibility of mouse testicular reproductive function impaired by single heat stress. Methods： Twenty-four C57 male mice were randomly divided into heat stress and control groups and immersed in water at 43℃ and 25℃,respectively,for 15 minutes. At 1,7,and 14 days of heat exposure,all the mice were sacrificed and their testis tissues collected for determining the apoptosis of the germ cells by TUNEL and measuring the expression level of the pH2AX protein by immunohistochemistry and Western blot. Results： The highest percentage of apoptotic cells were found in the seminiferous tubules of the mice in the heat stress group on the 1st day of the exposure and almost no apoptosis was observed at 7 and 14 days. The pH2AX protein was expressed in the nuclei of the basement membrane of adjacent seminiferous tubules. Compared with the control group,the expression of pH2AX was significantly increased on the 1st day of exposure（ 0. 47 ± 0. 02 vs 1. 61 ± 0. 04,P 〈0. 01）,then decreased at 7 days（ 0. 85 ± 0. 03） in comparison with that on the 1 st day（ P 〈0. 01）,and again elevated at 14 days（ 1. 72 ±0. 02） as compared with either those at 1 and 7 days（ P 〈0. 01） or that of the control（ P 〈0. 01）. Conclusion： Heat stress causes dynamic changes of the pH2AX expression in the testis of the mouse,which are associated with heat stress-induced proliferation and division of the testicular spermatogenic cells.

关 键 词：单次热应激 睾丸 pH2AX 生殖功能 可逆性 小鼠 

分 类 号：R698.2[医药卫生—泌尿科学]