隐睾发生过程中DNA甲基化状态和甲基化转移酶表达的实验研究 |
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引用本文: | 吴盛德,魏光辉,朱静,李娅莎,甘立强,袁心刚,徐明灯,陈柏林.隐睾发生过程中DNA甲基化状态和甲基化转移酶表达的实验研究[J].临床小儿外科杂志,2010,9(1):21-23,26. |
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作者姓名: | 吴盛德 魏光辉 朱静 李娅莎 甘立强 袁心刚 徐明灯 陈柏林 |
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作者单位: | 1. 重庆医科大学附属儿童医院泌尿外科,400014 2. 重庆医科大学附属儿科研究所,400014 |
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基金项目: | 国家自然科学基金,重庆市自然科学基金 |
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摘 要: | 目的研究环境内分泌干扰物邻苯二甲酸二(2-乙基已基)酯Di—(2-ethylhexy)phthalate,DEHP]作用于孕鼠后,胎鼠睾丸组织基因组DNA甲基化水平的变化及3种DNA甲基化转移酶的表达情况;探讨DEHP所致表观遗传修饰改变在睾丸下降过程中的作用。方法妊娠昆明小鼠(KMmice)随机分为3组,即正常对照组、玉米油对照组和DEHP实验组,每组15只。玉米油对照组和DEHP实验组自妊娠第12.5天到妊娠第18.5天分别持续经口予以玉米油和DEHP,用量按每日500mg/kg,正常对照组不予灌药。采用高效液相色谱仪检测基因组DNA甲基化水平,实时荧光定量PCR检测3种DNA甲基化转移酶的表达量。结果正常对照组和玉米油对照组DNA甲基化水平和3种甲基化转移酶的表达均无统计学意义(P〉0.05):DEHP实验组DNA甲基化水平比对照组增高约10%。与对照组相比,差异有显著统计学意义(P〈0.05);实验组3种甲基化转移酶的表达量均比对照组高,与对照组相比,差异有显著统计学意义(Dnmt1,P〈0.05;Dnmt3a,P〈0.01;Dnmt3b,P〈0.05)。结论环境内分泌干扰物DEHP通过影响3种甲基化转移酶的表达而导致基因组DNA甲基化水平升高,从而抑制睾丸下降过程中多种基因的表达,发生隐睾。DNA甲基化等表观遗传学修饰的改变可能是DEHP诱发隐睾的分子机制之一。
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关 键 词: | DNA甲基化 转移酶类 二乙基已基邻苯二甲酸/免疫学 隐睾 |
DNA methylation changes and DNA methyltransferases'expression during cryptorchidism formation induced by di-2-(ethylhexyl)phthalate in mice |
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Institution: | WU Sheng-de, WEI Guang-hui,ZHU Jing, et al.(1, Department of Urology, 2,Pediatric Institute, Children's Hospital of Chongqing Medical University, Chongqing 400014, China) |
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Abstract: | Objective To analyze epigenetic (specifically, DNA methylation) and DNA methyltransferases participation in the mechanisms of cryptorebidism only induced by maternal exposure to Di-2-(ethylhexyl) phthalate (DEHP) in mice. Methods KM miees were divided into 3 groups randomly: normal control group, corn oil group and DEHP-exposed group. Corn oil and DEHP 500 mg/ (kg· d)] were administrated respectively to the pregnant mice from gestation day 12.5 (GD12.5) to GD18.5. Testes were excised for analysis at GD19. High-performance liquid chromatography (HPLC) was performed to analyze DNA methylation status. The expression levels of the DNA methyhransferases were examined by Quantitative real-time PCR (qPCR). Results Using HPLC, at GD19, the status of DNA methylation in global DNA was increased significantly in DEHP-treated groups compared with the controls (P 〈 0.05). There were also significant changes in the expression of DNA methyhransferases (Dnmtl ,P 〈 0.05;Dnmt3a,P 〈 0.01 ;Dumt3b,P 〈 0.05). Conclusions These results indicate that changes in DNA methylation may play an important role in the manifestation of cryptorchidism only caused by environmental factors such as maternal exposure to DEHP. |
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Keywords: | DNA Methylation Transferases Diethylhexyl Phthalate/IM Cryptorehidism |
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