本文選題：吸煙　切入點(diǎn)：精子發(fā)生　出處：《上海交通大學(xué)》2012年碩士論文

【摘要】：吸煙被認(rèn)為是影響精子可育性的重要危險(xiǎn)因素,目前尚不清楚吸煙對(duì)精子可育性的影響機(jī)制。蛋白質(zhì)組學(xué)是研究人員認(rèn)識(shí)細(xì)胞功能最直接的工具,特定蛋白分子的鑒定能夠幫助我們認(rèn)識(shí)吸煙對(duì)精子不育影響的分子機(jī)制。 本研究首先通過(guò)睪丸的HE染色和精子數(shù)量、精子運(yùn)動(dòng)等參數(shù)的統(tǒng)計(jì)研究,發(fā)現(xiàn)吸煙后小鼠睪丸中精子數(shù)目明顯減少,生精小管之間的支持細(xì)胞數(shù)量較正常小鼠也明顯減少;附睪中分離的精子數(shù)量在吸煙后小鼠中明顯減少,精子的運(yùn)動(dòng)能力受吸煙的影響較大。這說(shuō)明了吸煙可能影響精子發(fā)生的功能,從而影響精子質(zhì)量。 為了進(jìn)一步探究吸煙對(duì)睪丸精子發(fā)生等功能影響的分子機(jī)制,我們通過(guò)比較蛋白質(zhì)組學(xué)技術(shù),發(fā)現(xiàn)了吸煙小鼠睪丸和正常小鼠睪丸之間差異表達(dá)的蛋白。我們用雙向凝膠電泳(2 - DE)和質(zhì)譜(MALDI - TOF - MS)技術(shù)鑒定吸煙小鼠睪丸和正常小鼠睪丸中差異表達(dá)的蛋白,發(fā)現(xiàn)了35種在吸煙小鼠睪丸中差異表達(dá)的蛋白,其中7種蛋白表達(dá)上調(diào),其他的28種蛋白表達(dá)下調(diào)。 為了分析鑒定這35種蛋白,我們選擇了DAVID服務(wù)器對(duì)他們聚類分析。這35種蛋白主要參與代謝,信號(hào)轉(zhuǎn)導(dǎo),細(xì)胞骨架組織和細(xì)胞運(yùn)動(dòng)維持等生物學(xué)過(guò)程。其中,在吸煙小鼠睪丸中表達(dá)下調(diào)的PEBP1可能與精子發(fā)生相關(guān)。我們分別用Western-Blot和qRT  PCR對(duì)PEBP1做了進(jìn)一步分析。PEBP1蛋白和mRNA表達(dá)量在吸煙小鼠睪丸中均有所下降,這與前面的雙向電泳結(jié)果一致。PEBP1是絲裂原激活的蛋白激酶(MAPK )信號(hào)轉(zhuǎn)導(dǎo)通路的生理性抑制蛋白,吸煙小鼠睪丸中PEBP1表達(dá)的下降,導(dǎo)致MAPK信號(hào)途徑的活躍。PEBP1可能成為臨床上評(píng)估吸煙對(duì)生殖不良影響的分子標(biāo)記,并提供吸煙對(duì)精子不育性的影響機(jī)制。
[Abstract]:Smoking is thought to be an important risk factor for sperm fertility, and it is not clear how smoking affects sperm fertility. Proteomics is the most direct tool for researchers to understand cell function. The identification of specific protein molecules can help us to understand the molecular mechanism of the effect of smoking on sperm sterility. In this study, the number of spermatozoa and Sertoli cells between seminiferous tubules were significantly decreased after smoking by means of HE staining, sperm number and sperm motility in testis of mice after smoking, and the number of Sertoli cells between seminiferous tubules was also significantly decreased compared with that of normal mice. The number of spermatozoa isolated from epididymis decreased significantly after smoking and the motility of spermatozoa was influenced by smoking, which indicated that smoking might affect the function of spermatogenesis and thus the quality of sperm. To further explore the molecular mechanism of the effects of smoking on testicular spermatogenesis, we compared proteomics techniques. The differentially expressed proteins were found between the testis of smoking mice and normal mice. The differential expression proteins were identified by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MALDI-MS) in the testis of smoking mice and normal mice. 35 differentially expressed proteins were found in the testis of smoking mice, 7 of which were up-regulated and the other 28 were down-regulated. In order to analyze and identify the 35 proteins, we chose the DAVID server to cluster them. These 35 proteins are mainly involved in the biological processes of metabolism, signal transduction, cytoskeleton tissue and cell movement maintenance. The down-regulated expression of PEBP1 in the testis of smoking mice may be related to spermatogenesis. We further analyzed the expression of PEBP1 protein and mRNA in the testis of smoking mice by Western-Blot and qRT PCR. This is consistent with the previous results of two-dimensional electrophoresis. PEBP1 is a physiological inhibitor of mitogen-activated protein kinase (MAPK) signal transduction pathway, and the decrease of PEBP1 expression in the testis of smoking mice. The activation of MAPK signaling pathway. PEBP1 may be a molecular marker to evaluate the adverse effects of smoking on reproduction and provide a mechanism for the effect of smoking on sperm sterility.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

【引證文獻(xiàn)】

中國(guó)期刊全文數(shù)據(jù)庫(kù) 前1條

1 黃德倫;黃愉淋;付強(qiáng);張明;;蛋白質(zhì)組學(xué)在哺乳動(dòng)物睪丸和精子發(fā)生中的應(yīng)用[J];基因組學(xué)與應(yīng)用生物學(xué);2013年02期

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本文編號(hào)：1687138