冷冻保存对正常和不育症精子PH-20表达和凋亡的影响

doi:10.16098/j.issn.0529-1356.2017.01.015

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 • 上一篇下一篇

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

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

收稿日期:2016-05-09 修回日期:2016-08-26 出版日期:2017-02-06 发布日期:2017-02-06
通讯作者: 高晓勤 E-mail:meryma0729@163.com
基金资助:
贵州省科技创新人才团队项目

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

1. Department of Histology and Embryology, Guizhou Medical University, Guiyang 550004,China;2. Department of Histology and Embryology, Zunyi Medical and Pharmaceutical College, Guizhou Zunyi 563006,China;3. Reproductive Medicine Center, Affiliated Hospital of Guizhou Medical University, Guiyang 550004,China;4. Clinical Experience Center, Affiliated Hospital of Guizhou Medical University, Guiyang 550004,China

Received:2016-05-09 Revised:2016-08-26 Online:2017-02-06 Published:2017-02-06
Contact: GAO Xiao-qin E-mail:meryma0729@163.com

摘要/Abstract

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。

关键词: 精子, 冷冻保存, PH-20, 免疫印迹法, 人

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.

Key words: Sperm, Cryopreservation, PH-20, Western blotting, Human

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.

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.

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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

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