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邻苯二甲酸二(2-乙基己基)酯增加小鼠睾丸基因组的DNA甲基化水平

[Di-(2-ethylhexyl) phthalate increases the DNA methylation level of genomes in the mouse testis].

作者信息

Wu Sheng-de, Zhu Jing, Li Ya-sha, Gan Li-qiang, Yuan Xin-gang, Xu Ming-deng, Wei Guang-hui

机构信息

Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.

出版信息

Zhonghua Nan Ke Xue. 2009 Oct;15(10):876-81.

Abstract

OBJECTIVE

To investigate the effect of the exposure to di- (2-ethylhexyl) phthalate (DEHP) during pregnancy on the DNA methylation level of genomes in the testis of the offspring in mice.

METHODS

Pregnant KM mice were randomly divided into three groups, normal control, corn oil and DEHP-exposed. Corn oil and DEHP (500 mg/[kg x d]) were administrated respectively from gestation day 12.5 (GD 12.5) to postnatal day 3 (PND 3). The testes of the offspring were excised on PND 7, and their genomic DNA was treated with EcoR I /Msp I and EcoR I /Hpa II. The genome-wide DNA methylation patterns of the CCGG sites were detected by methylation-sensitive amplification polymorphism (MSAP). The samples were electrophoresed in the ABI 3730 DNA sequencer and the results analyzed by the Genescan3.1.

RESULTS

The average incidence of DNA methylation was (34.03 +/- 3.05)% in the DEHP-exposed mice, obviously higher than (28.37 +/- 2.37)% in the normal control and (28.58 2.45)% in the corn oil group, with statistically significant differences (P < 0.05).

CONCLUSION

Exposure to DEHP during pregnancy increases the DNA methylation level of the genome in the testis of the offspring and affects the apparent genetic modification of the genome, which may be one of the important toxicological causes of the lesion in the reproductive system.

摘要

目的

研究孕期暴露于邻苯二甲酸二(2-乙基己基)酯(DEHP)对小鼠子代睾丸基因组DNA甲基化水平的影响。

方法

将怀孕的KM小鼠随机分为三组,正常对照组、玉米油组和DEHP暴露组。从妊娠第12.5天(GD 12.5)至出生后第3天(PND 3)分别给予玉米油和DEHP(500 mg/[kg×d])。在出生后第7天切除子代小鼠的睾丸,并用EcoR I /Msp I和EcoR I /Hpa II处理其基因组DNA。通过甲基化敏感扩增多态性(MSAP)检测CCGG位点的全基因组DNA甲基化模式。样本在ABI 3730 DNA测序仪上进行电泳,结果用Genescan3.1分析。

结果

DEHP暴露组小鼠DNA甲基化的平均发生率为(34.03±3.05)%,明显高于正常对照组的(28.37±2.37)%和玉米油组的(28.58±2.45)%,差异有统计学意义(P<0.05)。

结论

孕期暴露于DEHP会增加子代睾丸基因组的DNA甲基化水平,并影响基因组的表观遗传修饰,这可能是生殖系统损伤的重要毒理学原因之一。

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