冷冻保存对正常和不育症精子PH-20表达和凋亡的影响

马晓萍 高晓勤* 周桦 王珺

doi:10.16098/j.issn.0529-1356.2017.01.015

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解剖学报 ›› 2017, Vol. 48 ›› Issue (1) : 87-91. DOI: 10.16098/j.issn.0529-1356.2017.01.015

组织学胚胎学发育生物学

冷冻保存对正常和不育症精子PH-20表达和凋亡的影响

马晓萍^1,2 高晓勤^2* 周桦³ 王珺⁴

作者信息 +

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

MA Xiao-ping ^1,2 GAO Xiao-qin^2* ZHOU Hua³WANG Jun⁴

Author information +

文章历史 +

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(TdT)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。

Abstract

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(TdT) 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.

导出引用

马晓萍高晓勤周桦王珺. 冷冻保存对正常和不育症精子PH-20表达和凋亡的影响[J]. 解剖学报. 2017, 48(1): 87-91 https://doi.org/10.16098/j.issn.0529-1356.2017.01.015

MA Xiao-ping GAO Xiao-qin ZHOU Hua WANG Jun. Effect of cryopreservation on the expression of sperm surface protein PH-20 and ratio of apoptosis in normal birth and infertility[J]. Acta Anatomica Sinica. 2017, 48(1): 87-91 https://doi.org/10.16098/j.issn.0529-1356.2017.01.015

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