冷冻保存对正常和不育症精子PH-20表达和凋亡的影响  

Effect of cryopreservation on the expression of sperm surface protein PH-20 and ratio of apoptosis in normal birth and infertility

作　　者：马晓萍[1,2] 高晓勤[2] 周桦[3] 王珺[4] 

机构地区：[1]贵州医科大学组织学与胚胎学教研室,贵阳550004 [2]遵义医药高等专科学校组织学胚胎学教研室,贵州遵义563006 [3]贵州医科大学附属医院生殖医学中心,贵阳550004 [4]贵州医科大学附属医院临床实验中心,贵阳550004

出　　处：《解剖学报》2017年第1期87-91,共5页Acta Anatomica Sinica

基　　金：贵州省科技创新人才团队项目[黔科合人才团队(2014)4005号];贵州省科技计划项目[黔科合LH字(2015)7567号]

摘　　要：目的探讨冷冻保存对人精子膜蛋白PH-20表达和精子凋亡的影响。方法 14例正常生育力精液标本(A组)和20例不育症精液标本(B组)行冷冻保存。Western blotting检测PH-20蛋白在人精子中的表达,免疫荧光用来观察PH-20蛋白在人精子上的定位,应用末端脱氧核苷酸转移酶(Td T)介导的原位末端标记(TUNEL)法检测精子凋亡情况。结果解冻后正常生育组和不育组的PH-20/β-actin平均吸光度与冷冻前比较均有显著性下降(P<0.05);解冻后正常生育组和不育组的PH-20阳性率与冷冻前比较均有显著性下降(P<0.05)。解冻后正常生育组的精子凋亡率与冷冻前的比较差异无显著性意义(P>0.05)。而解冻后不育组的精子凋亡率与冷冻前的比较均显著性下降(P<0.05),且不育组的降低程度大于正常生育组。结论冷冻-解冻过程可引起精子PH-20蛋白表达减少和精子PH-20阳性率降低,但冷冻保存对正常生育者的精子凋亡率无显著影响。Objective To investigate the effect of cryopreservation on the expression of sperm surface protein PH-20 and the sperm apoptosis.Methods Semen samples were obtained from fertile men（n = 14,group A） and infertile men（n = 20,group B）.Western blotting was used to detect the PH-20 protein expression in human spermatozoa.The localization of this protein on human spermatozoa was determined by indirect immunofluorescent staining using PH-20 antibody.The sperm apoptosis was examined by terminal deoxynucleotidyl transferase（Td T） mediated deoxyuridine triphophate-biotin nick end labeling（TUNEL）.Results After cryopreservation,the level of PH-20 protein expression was significantly lower in both group A and B than that of fresh sperm（P 〈 0.05）.The percentage of PH-20 positive rate was significantly lower in both group A and B than that of fresh sperm（P 〈 0.05）.The ratio of sperm apoptosis was not significantly different in thawed group A than that of fresh sperm（P 〉 0.05）.The ratio of sperm apoptosis was significantly lower in thawed group B than that of fresh sperm（P 〈 0.05）.Conclusion Cryopreservation caused significant reduction of PH-20 protein expression,the percentage of PH-20 positive rate in human sperm.But the cryopreservation had no significant influence on normal fertility of sperm apoptosis.

关 键 词：精子 冷冻保存 PH-20 免疫印迹法 人 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]