發(fā)布時間：2019-06-26 18:53

【摘要】：目的:本文擬通過建立母鼠孕期暴露于交通性污染物染毒模型,研究孕鼠暴露于交通性污染物后對其子代生殖系統(tǒng)的影響。同時借助簡化表觀亞硫酸氫鹽測序技術(shù)從表觀遺傳學的角度分析交通性污染物對子代雄性生殖系統(tǒng)的影響機制,揭示交通性污染物導致雄性生育能力下降的作用靶點和機理,為尋找男性生殖功能降低的原因和治療男性不育癥提供實驗與理論依據(jù)。方法:30只孕鼠(C57BL/6J種系)隨機分為對照組和暴露組,每組各15只。暴露組孕鼠置于重慶市主城區(qū)某一車流量較大的隧道內(nèi),從D4-D14每天持續(xù)暴露8h。其余時間暴露組與對照組孕鼠均分籠飼養(yǎng)于重慶醫(yī)科大學動物實驗中心普通級,溫度(22±1)℃,相對濕度40%-60%,自由進食和飲水。D18后將孕鼠單籠喂養(yǎng)直至其自然分娩。子代雄性小鼠出生后正常飼養(yǎng)至性成熟(55日齡)。在暴露組染毒的D4、D8、D12、D14對隧道交通性污染物NOX、TSP、PM2.5、PM10和噪聲進行采集和檢測。染毒結(jié)束后觀察孕鼠流產(chǎn)情況,子代小鼠生殖系統(tǒng)臟器系數(shù),小鼠精子數(shù)量、畸形率、小鼠睪丸組織病理學變化。兩組各取3只雄性小鼠睪丸進行RRBS甲基化測序并用Real-time PCR、Western Bolt方法檢測目的基因Rpe、Aldh7a1的m RNA和蛋白表達水平。結(jié)果:隧道交通性污染物NOX(0.079±0.147 mg/m3)、TSP(0.61±0.049 mg/m3)、PM2.5(141±23.45 mg/m3)、PM10(206±20.78 mg/m3)和噪聲[77.60±7.05 d B(A)]均超過對照組,差異有統(tǒng)計學意義(P0.05),兩組SO2濃度差異沒有顯著性(P0.05)。暴露組孕鼠流產(chǎn)率(66.67%)高于對照組。暴露組雄鼠精子數(shù)量(9.70±5.54′107/g)少于對照組(12.77±2.72′107/g),差異有統(tǒng)計學意義(P0.05),其體質(zhì)量、睪丸和附睪臟器系數(shù)、精子畸形率與對照組相比,無統(tǒng)計學改變(P0.05)。電鏡結(jié)果顯示,暴露組雄鼠睪丸組織中生精小管之間空隙大,管腔內(nèi)出現(xiàn)較多空泡。與對照組相比,暴露組子代雄鼠睪丸基因組有34個基因上調(diào),81個基因下調(diào)。Rpe、Aldh7a1基因分別調(diào)控糖酵解途徑和丙酮酸代謝過程。與對照組相比暴露組Rpe、Aldh7a1基因m RNA和蛋白相對表達量減少(P0.05)。結(jié)論:孕鼠交通性污染物暴露可使子代雄鼠精子數(shù)量下降、睪丸組織形態(tài)學發(fā)生一定程度的改變,也可引起子代雄鼠生殖系細胞表觀基因組發(fā)生改變。Rpe、Aldh7a1基因下調(diào)而導致的氨基酸合成通路受到影響可能是導致子代雄鼠生殖系統(tǒng)損害的原因之一。
[Abstract]:Aim: to establish a model of maternal exposure to traffic pollutants during pregnancy and to study the effects of maternal exposure to traffic pollutants on the reproductive system of their offspring. At the same time, with the help of simplified apparent bisulfite sequencing technique, the influence mechanism of traffic pollutants on male reproductive system of offspring was analyzed from the point of view of epigenetics, and the target and mechanism of male fertility decline caused by traffic pollutants were revealed, which provided experimental and theoretical basis for finding out the causes of male reproductive function decline and treating male sterility. Methods: 30 pregnant rats (C57BL/6J strain) were randomly divided into control group (n = 15) and exposure group (n = 15). The pregnant rats in the exposure group were exposed to D4-D14 for 8 hours per day in a tunnel with large traffic flow in the main urban area of Chongqing. For the rest of the time, the pregnant rats in the exposure group and the control group were fed in cages in the animal experimental center of Chongqing Medical University, the temperature was (22 鹵1) 鈩，

本文編號：2506403