本文選題：雙酚A + 睪丸生殖細胞　； 參考：《安徽醫(yī)科大學》2013年碩士論文

【摘要】：背景：雙酚A(bisphenol A, BPA)是環(huán)境雌激素中最常見的一種,屬于全球性的環(huán)境污染物,隨著科技的發(fā)展而應用到日常生活中的很多領域。研究表明BPA的半衰期較長,對機體的生殖系統(tǒng)、免疫系統(tǒng)、神經(jīng)系統(tǒng)和胚胎發(fā)育等方面都有不利的影響,甚至可使機體發(fā)生癌變等,嚴重威脅著人類及后代的健康。已有實驗表明BPA對機體的生殖功能產(chǎn)生不可逆的破壞作用,特別是對雄性生殖系統(tǒng)的發(fā)育。然而孕期BPA暴露對子代雄性生殖毒性仍然有不同的看法。 目的：本研究觀察BPA暴露對子鼠雄性生殖細胞的形態(tài)學變化和細胞凋亡的影響,進一步探討B(tài)PA對雄性生殖系統(tǒng)的影響及可能機制。 方法：ICR孕鼠隨機分為空白對照組、溶劑對照組、BPA低劑量組(10nmol/L)、BPA中劑量組(100nmol/L)和BPA高劑量組(1000nmol/L),每組8只；孕鼠飲水染毒至小鼠斷乳后42d[出生后天數(shù)(postnatal day, PND42)],于PND21和PND42分別處死雄性子鼠。進行以下研究：(1)統(tǒng)計每組每只孕鼠生產(chǎn)子鼠的數(shù)目及雌雄比例；計算睪丸臟器系數(shù)(睪丸濕重與子鼠體重比值)；(2)H.E染色觀察子鼠睪丸組織的形態(tài)學變化；(3)透射電鏡觀察睪丸組織電鏡改變；(4)Hoechst33258染色熒光顯微鏡檢測觀察子鼠睪丸生殖細胞凋亡情況；(5)免疫組化染色SABC法檢測子鼠睪丸生殖細胞Caspase-3的表達情況；(6)Western blot檢測子鼠睪丸組織勻漿Caspase-3的表達量。 結(jié)果：各組每只母鼠的子鼠數(shù)量和雌雄比例與對照組相比差異無顯著性(P0.05)。隨著BPA染毒劑量的增加,PND21和PND42的實驗組小鼠的睪丸臟器系數(shù)呈下降趨勢,以中、高劑量組更為明顯。H.E染色顯示,BPA暴露組子鼠睪丸生精小管管壁細胞層數(shù)減少,細胞排列零亂,其中6W的中、高劑量組較為明顯；電鏡結(jié)果顯示BPA中、高劑量組小鼠睪丸內(nèi)精原細胞、支持細胞和間質(zhì)細胞呈現(xiàn)病理學改變,精原細胞、支持細胞和間質(zhì)細胞內(nèi)的線粒體嵴消失呈明顯空泡樣改變,雙層膜結(jié)構(gòu)不清,粗面內(nèi)質(zhì)網(wǎng)出現(xiàn)不同程度的擴張,結(jié)構(gòu)零亂松散,且有些細胞線粒體數(shù)目減少,細胞核染色質(zhì)邊集；Hoechst33258染色結(jié)果示：中劑量組精原細胞、支持細胞和間質(zhì)細胞的核呈致密濃染,使細胞核呈現(xiàn)高強度的固縮狀態(tài)的藍色熒光,即細胞發(fā)生凋亡；Western blot和免疫組化結(jié)果均顯示：BPA中、高劑量組子鼠睪丸生殖細胞Caspase-3的表達增加,且多位于睪丸間質(zhì)細胞中和靠近基膜的精原細胞和支持細胞中。 結(jié)論：出生前后暴露BPA可使小鼠的睪丸臟器系數(shù)下降,生精細胞、支持細胞和間質(zhì)細胞發(fā)生形態(tài)學改變和細胞凋亡增加等,說明BPA可能通過胎盤屏障和母乳影響雄性子鼠生殖系統(tǒng)的發(fā)育,其機制可能與增加雄性子鼠生殖細胞的凋亡有關。
[Abstract]:Background: bisphenol A (BPAA) is one of the most common environmental estrogens, which is a global environmental pollutant. With the development of science and technology, it has been applied in many fields of daily life. The results show that BPA has a long half-life, which has adverse effects on the reproductive system, immune system, nervous system and embryonic development of the body, and even makes the body cancerous, which seriously threatens the health of human beings and their offspring. It has been shown that BPA has irreversibly destructive effects on reproductive function, especially on the development of male reproductive system. However, BPA exposure during pregnancy still has different views on male reproductive toxicity in offspring. Aim: to observe the effects of BPA exposure on the morphological changes and apoptosis of male germ cells in rats and to explore the effects of BPA on the male reproductive system and its possible mechanism. Methods Twenty ICR pregnant mice were randomly divided into two groups: control group (n = 10 nmol / L) and high dose BPA group (n = 8). The pregnant mice were exposed to drinking water until 42 days after weaning (postnatal day (PND 42), and the male mice were killed at PND21 and PND42, respectively. Do the following research: 1) count the number and the ratio of male and female to each group of pregnant rats; Calculation of testicular visceral coefficient (ratio of wet weight of testis to body weight of the offspring) the morphological changes of testis were observed by means of electron microscopy (transmission electron microscope (TEM) observation of testicular tissue changes by fluorescence microscopy with Hoechst33258 staining The expression of Caspase-3 in testicular germ cells was detected by immunohistochemical SABC method. The expression of Caspase-3 in testis homogenate was detected by Western blot. Results: there was no significant difference in the number and ratio of female and male between each group and the control group (P 0.05). With the increase of the dose of BPA, the testicular viscera coefficient of mice in PND21 and PND42 groups showed a decreasing trend. In the middle and high dose groups, the number of testicular seminiferous tubule wall cells decreased and the cells arranged in disorder in the middle and high dose groups. The results of electron microscope showed that the spermatogonium, Sertoli cells and interstitial cells in the testis of BPA mice showed pathological changes. In Sertoli cells and interstitial cells, the mitochondrial cristae disappeared with obvious vacuolar changes, the double layer membrane structure was unclear, the rough endoplasmic reticulum expanded to varying degrees, the structure was loose, and the number of mitochondria in some cells decreased. The results of Hoechst33258 staining showed that the nuclei of Sertoli cells and interstitial cells in the middle dose group were dense and dense, resulting in a high intensity blue fluorescence of the nucleus in the pyknotic state. The results of Western blot and immunohistochemistry showed that the expression of Caspase-3 in testis germ cells increased in high dose group, and most of them were located in stromal cells of testis and spermatogonia and Sertoli cells near the basement membrane. Conclusion: exposure to BPA before and after birth can decrease the testicular organ coefficient and increase the morphological changes and apoptosis of spermatogenic cells Sertoli cells and interstitial cells in mice. It is suggested that BPA may influence the development of reproductive system of male offspring through placental barrier and breast milk, and its mechanism may be related to increasing the apoptosis of germ cells in male offspring.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R114

【參考文獻】

相關期刊論文 前8條

1 張鏇;蔣學武;黃天華;;國人先天性尿道下裂的病因?qū)W研究進展[J];癌變.畸變.突變;2008年01期

2 宋清坤;郝衛(wèi)東;尚蘭琴;魏雪濤;蔣建軍;邢麗娜;王輝;吳雙;趙燕;喬楊崢;郝長付;;雙酚A對青春期雄性大鼠生殖發(fā)育的影響[J];癌變.畸變.突變;2008年04期

3 逄兵,周袁芬,周天喜,任道風,金泰^，

本文編號：1964407