【摘要】：目的探討冷凍保存對(duì)人精子膜蛋白PH-20表達(dá)和精子凋亡的影響。方法 14例正常生育力精液標(biāo)本(A組)和20例不育癥精液標(biāo)本(B組)行冷凍保存。Western blotting檢測(cè)PH-20蛋白在人精子中的表達(dá),免疫熒光用來(lái)觀察PH-20蛋白在人精子上的定位,應(yīng)用末端脫氧核苷酸轉(zhuǎn)移酶(Td T)介導(dǎo)的原位末端標(biāo)記(TUNEL)法檢測(cè)精子凋亡情況。結(jié)果解凍后正常生育組和不育組的PH-20/β-actin平均吸光度與冷凍前比較均有顯著性下降(P0.05);解凍后正常生育組和不育組的PH-20陽(yáng)性率與冷凍前比較均有顯著性下降(P0.05)。解凍后正常生育組的精子凋亡率與冷凍前的比較差異無(wú)顯著性意義(P0.05)。而解凍后不育組的精子凋亡率與冷凍前的比較均顯著性下降(P0.05),且不育組的降低程度大于正常生育組。結(jié)論冷凍-解凍過(guò)程可引起精子PH-20蛋白表達(dá)減少和精子PH-20陽(yáng)性率降低,但冷凍保存對(duì)正常生育者的精子凋亡率無(wú)顯著影響。
[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.
【作者單位】： 貴州醫(yī)科大學(xué)組織學(xué)與胚胎學(xué)教研室;遵義醫(yī)藥高等�？茖W(xué)校組織學(xué)胚胎學(xué)教研室;貴州醫(yī)科大學(xué)附屬醫(yī)院生殖醫(yī)學(xué)中心;貴州醫(yī)科大學(xué)附屬醫(yī)院臨床實(shí)驗(yàn)中心;
【基金】：貴州省科技創(chuàng)新人才團(tuán)隊(duì)項(xiàng)目[黔科合人才團(tuán)隊(duì)(2014)4005號(hào)] 貴州省科技計(jì)劃項(xiàng)目[黔科合LH字(2015)7567號(hào)]
【分類號(hào)】：R698.2

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