冷冻保存对正常和不育症精子PH-20表达和凋亡的影响

发布时间：2019-03-07 10:42

【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(P0.05);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(P0.05)。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(P0.05)。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(P0.05),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 human sperm membrane protein PH-20 and sperm apoptosis. Methods 14 normal semen samples (group A) and 20 infertile semen samples (group B) were treated with cryopreserved. Western blotting to detect the expression of PH-20 protein in human spermatozoa. Immunofluorescence was used to observe the localization of PH-20 protein in human spermatozoa. Apoptosis of spermatozoa was detected by terminal deoxynucleotidyl transferase (Td T) mediated in situ end labeling (TUNEL). Results the average absorbance of PH-20/ 尾-actin in normal fertility group and infertile group after thawing was significantly lower than that before freezing (P0.05). The positive rate of PH-20 in normal fertility group and infertile group after thawing was significantly lower than that before freezing (P0.05). There was no significant difference in sperm apoptosis rate between normal fertility group after thawing and before freezing (P0.05). After thawing, the sperm apoptosis rate in the infertile group was significantly lower than that before freezing (P0.05), and the degree of decrease in the infertile group was greater than that in the normal fertility group. Conclusion freezing-thawing process can decrease the expression of PH-20 protein and the positive rate of PH-20 in spermatozoa, but cryopreservation has no significant effect on sperm apoptosis rate.
【作者单位】：贵州医科大学组织学与胚胎学教研室;遵义医药高等专科学校组织学胚胎学教研室;贵州医科大学附属医院生殖医学中心;贵州医科大学附属医院临床实验中心;
【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]
【分类号】：R698.2

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