【摘要】：目的精子活力低下所導(dǎo)致的弱精癥是男性不育的重要原因之一，深入探討弱精癥的發(fā)生機(jī)制、提出新的治療策略是國(guó)內(nèi)外生殖領(lǐng)域的一個(gè)研究熱點(diǎn)。本實(shí)驗(yàn)旨在通過(guò)測(cè)定弱精癥精子細(xì)胞的活力、DNA損傷率及中心小體蛋白135（centrosomalprotein135kDa，，CEP135）的相關(guān)表達(dá)量，并探討CEP135表達(dá)與精子活力和精子DNA損傷率之間的相互關(guān)系，以確定CEP135在弱精癥診療過(guò)程中的指導(dǎo)價(jià)值，并對(duì)其作用機(jī)制進(jìn)行初步探討。 方法應(yīng)用計(jì)算機(jī)輔助精液分析對(duì)91例精液樣本進(jìn)行參數(shù)檢測(cè)，按照WHO標(biāo)準(zhǔn)，根據(jù)（a+b）級(jí)精子百分率分為3組，Ⅰ組31例：（a+b）級(jí)精子＞50%或a級(jí)精子＞25%；Ⅱ組30例：（a+b）級(jí)精子25%~50%；Ⅲ組30例：（a+b）級(jí)精子＜25%。利用精子染色質(zhì)擴(kuò)散實(shí)驗(yàn)（Sperm chromatin dispersion test,SCD）檢測(cè)各組精子細(xì)胞的DNA損傷率。逆轉(zhuǎn)錄-實(shí)時(shí)定量聚合酶鏈反應(yīng)（Reversetranscription-quantitative real-time PCR， RT-qPCR）檢測(cè)CEP135信使RNA（Messenger RNA，mRNA）在三組精液中的表達(dá)情況，采用2-ΔCT公式（ΔCT=CT目的-CT內(nèi)參）計(jì)算目的基因的相對(duì)表達(dá)量（Relative expression，Re），瓊脂糖凝膠電泳驗(yàn)證RT-qPCR的實(shí)驗(yàn)結(jié)果。應(yīng)用SPSS16.0軟件統(tǒng)計(jì)數(shù)據(jù)并對(duì)精子細(xì)胞的活力、DNA損傷率及CEP135mRNA的相對(duì)表達(dá)量進(jìn)行相關(guān)性分析。 結(jié)果3組精液中精子細(xì)胞的活力分別為62.31%±7.97%、36.63%±6.61%、16.51%±6.57%，DNA損傷率分別為17.18%±6.72%、22.86%±6.67%、27.30%±6.44%，CEP135mRNA的平均相對(duì)表達(dá)量分別為9.45×10-2±2.64×10-4、9.48×10-2±3.24×10-4、9.49×10-2±2.52×10-4，各組之間比較差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）。DNA損傷率及CEP135mRNA的相對(duì)表達(dá)量均與精子細(xì)胞的活力呈顯著負(fù)相關(guān)（r分別為-0.619，-0.574；P均＜0.01）；隨著CEP135mRNA相對(duì)表達(dá)量的升高，DNA損傷率也逐漸增加，二者之間顯著正相關(guān)（r=0.316；P＜0.01）。 結(jié)論弱精癥精液中高表達(dá)的CEP135mRNA與精子細(xì)胞活力的降低及DNA損傷率的增高密切相關(guān)，具有顯著相關(guān)性。臨床上可以通過(guò)測(cè)定精子細(xì)胞CEP135mRNA的相對(duì)表達(dá)量來(lái)評(píng)估患者的精液質(zhì)量，并通過(guò)基因干擾技術(shù)影響CEP135mRNA的相對(duì)表達(dá)量進(jìn)而改善精液質(zhì)量，提高弱精癥的生育能力。
[Abstract]:Objective Aspermia caused by low sperm motility is one of the important causes of male infertility. It is a hot research topic in reproductive field at home and abroad to probe into the mechanism of spermatogenesis and put forward new treatment strategy. The aim of this study was to investigate the relationship between CEP135 expression and sperm motility and sperm DNA damage rate by measuring the activity of spermatozoa, the rate of DNA damage, and the expression of central body protein 135 (centrosomalprotein135kDa,CEP135) in asthenospermic spermatozoa. To determine the guiding value of CEP135 in the diagnosis and treatment of azoospermia and to explore its mechanism. Methods the parameters of 91 semen samples were measured by computer assisted semen analysis. According to the WHO standard, according to the percentage of (a b) grade sperm, 31 cases of group 鈪

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