氢化可的松对保存过程中猪精子的影响  

Effect of hydrocortisone on preservation of boar sperm

作　　者：孙欢欢 王娜 孙良振 郭海涛 王婧然 杨康 周佳勃 谭景和 SUN Huan-huan;WANG Na;SUN Liang-zhen;GUO Hai-tao;WANG Jing-ran;YANG Kang;ZHOU Jia-bo;TAN Jing-he(Heilongjiang Key Laboratory of Animal Cell and Genetic Engineering,College of Life Sciences,Northeast Agricultural University,Harbin 150030,China;College of Animal Science and Tech nology,Shandong Agricultural University,Tai an,Shan-dong 271018,China)

机构地区：[1]东北农业大学生命科学学院黑龙江省动物细胞与遗传工程重点实验室,黑龙江哈尔滨150030 [2]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2020年第2期405-409,417,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31372311);东北农业大学研究基金资助项目(106518001)。

摘　　要：动物长期处于应激状态会影响其生育能力。应激对雄性动物生殖影响的研究多在体内进行,但在体外研究甚少,因此本试验在体外添加糖皮质激素研究其对猪精子各项生理指标的影响。试验首先在液态保存中分别添加不同质量浓度的氢化可的松(0.1,0.25,0.5,10.0,50.0 mg/L),检测其对猪精子活力的影响。结果显示,从0.25 mg/L应激质量浓度氢化可的松开始精子活力逐渐下降且均低于对照组(P<0.05)。之后通过蛋白质印迹法在射出的猪精子中证明存在糖皮质激素受体(glucocorticoid receptor,GR)。免疫荧光检测表明GR主要在精子顶体后区及尾部中段表达。精子保存第5天时,氢化可的松+米非司酮处理组精子活力显著高于氢化可的松单独处理组(P<0.05)。随后又检测了长期(5 d)处于应激质量浓度氢化可的松作用下,精子DNA碎裂(DFI)、线粒体膜电位(MMP)、精子凋亡水平。结果表明,精子DFI增加,线粒体功能下降,凋亡比例增加,而这些作用可以被米非司酮拮抗。因此表明氢化可的松可能是通过与其受体GR结合干扰线粒体功能而影响猪精子。Long-term stress in animals can affect their fertility.The research on the effects of stress on male reproductive performance was mostly carried out in vivo;however,few in vitro studies have been reported.Therefore,this experiment added glucocorticoid in vitro to study its effects on various physiological parameters of boar sperm.Firsty,the effects of different concentrations of hydrocortisone(0.1,0.25,0.5,10.0,50.0 mg/L) on sperm preservation were evaluated by sperm motility.The results showed that sperm motility gradually decreased from hydrocortisone at a stress concentration of 0.25 mg/L and was lower than that of the control group(P<0.05).The presence of glucocorticoid receptor(GR) was then demonstrated in the ejected boar sperm by Western blot.Immunofluorescence technique showed that GR was mainly expressed in the postacrosomal region and midpiece in the boar sperm.On the 5 th day of sperm preservation,the sperm motility of hydrocortisone+mifepristone treatment group was significantly higher than that of hydrocortisone alone treatment group.Subsequently,the levels of sperm DNA fragmentation(DFI),mitochondrial membrane potential(MMP) and sperm apoptosis were measured after exposure to hydrocortisone for a long period of time(5 days).The results showed that sperm DFI increased,mitochondrial function decreased and apoptotic rate increased.These effects could be antagonized by mifepristone.It was therefore suggested that hydrocortisone might affect boar sperm by binding to its receptor GR to interfere with mitochondrial function.

关 键 词：猪精液 液态保存 糖皮质激素受体 氢化可的松 米非司酮 

分 类 号：S84[农业科学—畜牧兽医] S852