本文選題：Igf + 甲基化��； 參考：《環(huán)境與職業(yè)醫(yī)學(xué)》2017年02期

【摘要】：[目的]建立宮內(nèi)p,p’-DDE暴露跨代大鼠模型,觀察雄性子代生殖毒性是否具有親源性遺傳特征及可能的表觀遺傳機制。[方法]SPF級的SD大鼠于孕8~15天給予每天每千克體重100 mg p,p’-DDE灌胃,另設(shè)溶劑對照組采用玉米油灌胃,孕鼠自由分娩,保留所有雄性仔鼠(F1代),同組別不同窩別的F1代仔鼠于出生后90天按雌雄比1∶1交配,產(chǎn)生F2代仔鼠。染毒組和對照組雌雄F2代按以下分組設(shè)計兩兩交配后,產(chǎn)生F3代仔鼠,分組為:(1)雌雄均為對照組(C♂-C♀),(2)雌雄均為染毒組(DDE♂-DDE♀),(3)染毒組雄性與對照組雌性(DDE♂-C♀),(4)對照組雄性與染毒組雌性(C♂-DDE♀)。用計算機輔助精子分析系統(tǒng)(CASA)分析3代雄性仔鼠成年后的精子質(zhì)量,亞硫酸氫鈉基因組測序法檢測Igf2 DMR2區(qū)域甲基化水平,實時熒光定量PCR分析印記基因表達。[結(jié)果]p,p’-DDE可誘導(dǎo)F1、F2及F3代DDE♂-DDE♀、DDE♂-C♀精子數(shù)量和活力下降,F1代精子數(shù)量和活力分別從(70.63±11.79)×106/m L、(76.75±7.57)%下降至(50.00±13.08)×106/m L、(62.42±9.40)%;F2代精子數(shù)量和活力分別從(82.86±38.60)×106/m L、(82.14±6.44)%下降至(43.75±25.04)×106/m L、(60.88±25.03)%;F3代DDE♂-DDE♀精子數(shù)量及活力分別從(68.89±41.37)×106/m L、(68.56±10.67)%下降至(21.66±4.83)×106/m L和(29.33±32.70)%,F3代DDE♂-C♀精子數(shù)量和活力下降為(28.00±16.60)×106/m L和(34.60±31.60)%,差異均有統(tǒng)計學(xué)意義(均P0.05)。F1~F3代雄性仔鼠精子細胞Igf2 DMR2區(qū)域(1、16、18位點)低甲基化,Igf2轉(zhuǎn)錄下調(diào),H19轉(zhuǎn)錄上調(diào),Igf2基因的轉(zhuǎn)錄水平和Igf2 DMR2甲基化成正相關(guān)(r=0.806 5)。[結(jié)論]Igf2 DMR2區(qū)域甲基化改變可能在p,p’-DDE誘導(dǎo)的跨代雄性大鼠生殖毒性中起重要作用。
[Abstract]:[objective] to establish an intergenerational rat model of intrauterine pSP-DDE exposure to observe whether the reproductive toxicity of male offspring has genetic characteristics and the possible epigenetic mechanism. [methods] Sprague-Dawley rats of SPF grade were given intragastric administration of 100mg / kg body weight per day for 815 days of gestation. The control group was fed with corn oil, and the pregnant rats gave birth freely. All the male offspring (F1 generation) were kept and the F1 offspring of the same group and different litter groups were mated at 1:1 after birth on the 90th day after birth to produce F2 offspring. The F _ 2 generation of the exposed group and the control group were designed for mating according to the following groups, and the offspring of the F _ 3 generation were produced. The two groups were as follows: (1) both male and female were control group (C-C), (_ 2). Both male and female were), (_ (3). The male and female of the control group and the control group (), (_ (4) were divided into two groups: the control group and the exposed female group (C ~ + -DDE). The sperm quality was analyzed by computer-aided sperm analysis system (CASA), the methylation level of Igf2D MR2 region was detected by genomic sequencing of sodium bisulfite, and the imprinted gene expression was analyzed by real-time fluorescence quantitative PCR. [緇撴灉]p,p鈥，

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