p,p’-DDE致大鼠跨代雄性生殖毒性效应及Igf2 DMR2区域甲基化的可能机制

发布时间：2018-06-22 11:19

  本文选题：Igf + 甲基化　； 参考：《环境与职业医学》2017年02期

【摘要】：[目的]建立宫内p,p’-DDE暴露跨代大鼠模型,观察雄性子代生殖毒性是否具有亲源性遗传特征及可能的表观遗传机制。[方法]SPF级的SD大鼠于孕8~15天给予每天每千克体重100 mg p,p’-DDE灌胃,另设溶剂对照组采用玉米油灌胃,孕鼠自由分娩,保留所有雄性仔鼠(F1代),同组别不同窝别的F1代仔鼠于出生后90天按雌雄比1∶1交配,产生F2代仔鼠。染毒组和对照组雌雄F2代按以下分组设计两两交配后,产生F3代仔鼠,分组为:(1)雌雄均为对照组(C♂-C♀),(2)雌雄均为染毒组(DDE♂-DDE♀),(3)染毒组雄性与对照组雌性(DDE♂-C♀),(4)对照组雄性与染毒组雌性(C♂-DDE♀)。用计算机辅助精子分析系统(CASA)分析3代雄性仔鼠成年后的精子质量,亚硫酸氢钠基因组测序法检测Igf2 DMR2区域甲基化水平,实时荧光定量PCR分析印记基因表达。[结果]p,p’-DDE可诱导F1、F2及F3代DDE♂-DDE♀、DDE♂-C♀精子数量和活力下降,F1代精子数量和活力分别从(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代精子数量和活力分别从(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♀精子数量及活力分别从(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♀精子数量和活力下降为(28.00±16.60)×106/m L和(34.60±31.60)%,差异均有统计学意义(均P0.05)。F1~F3代雄性仔鼠精子细胞Igf2 DMR2区域(1、16、18位点)低甲基化,Igf2转录下调,H19转录上调,Igf2基因的转录水平和Igf2 DMR2甲基化成正相关(r=0.806 5)。[结论]Igf2 DMR2区域甲基化改变可能在p,p’-DDE诱导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. [缁撴灉]p,p鈥，

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