大鼠卵巢镉暴露对DNA甲基转移酶表达的实验研究  

Effects of cadmium exposure on DNA methylation transferase in ovaries of postnatal rats

作　　者：吴停停[1,2] 谢美美[1] 罗凌凤[2] 卢晓丽[1] 林森斌[2] 陈洁[2] 张文昌[2] 

机构地区：[1]福建省疾病预防控制中心,福建福州350001 [2]福建医科大学公共卫生学院环境与健康重点实验室

出　　处：《中国预防医学杂志》2016年第6期401-404,共4页Chinese Preventive Medicine

基　　金：国家自然科学基金(81373027)

摘　　要：目的研究新出生卵巢组织体外镉暴露后,DNA甲基转移酶类(DNMTs)基因转录和蛋白表达的情况,了解DNMTs的变化在镉致卵巢毒性中的作用。方法 4日龄雌性SD大鼠卵巢取出置于DMEM/F12与α-MEM混合培养体系进行体外染毒,镉剂量分别为0、0.5、10、50μmol/L,连续染毒4d。分别采用Real Time-PCR和Westernblot方法检测DNMTs mRNA及其蛋白表达水平。应用单因素方差进行分析比较,P<0.05为差异有统计学意义。结果各剂量组DNMT1、DNMT3α、DNMT3βmRNA与对照组比较,差异均有统计学意义(F=13.579、12.926、10.620,P=0.009、0.012、0.031),其中DNMTs在0.5、10μmol/L组表达上调,50μmol/L组表达下调;各剂量组DNMT1蛋白表达与对照组比较,差异有统计学意义(F=56.311,P<0.01),其中在0.5μmol/L和50μmol/L组降低,10μmol/L组表达升高;DNMT3α蛋白表达在10μmol/L组升高,50μmol/L组降低,与对照组比较差异有统计学意义(F=28.533,P<0.01);各剂量组DNMT3β蛋白表达与对照组比较差异无统计学意义(F=2.047,P=0.186)。结论在本次实验条件下,镉暴露后卵巢组织DNMTs表达发生改变,这可能参与镉致卵巢毒性过程。Objective To observe the effects of cadmium exposure on DNA methytransferases（DNMTs）in ovaries of postnatal rats in vitro. Methods The ovaries of Sprague-Dawley rats were harvested on postnatal day4（PND4）,and then cultured in the medium mixed with DMEM/F12andα-minimal essential medium（α-MEM）at 1∶1ratio,which contained different concentration of CdCl2（0,0.5,5,10,50μmol/L）.After 4days of incubation,total RNA and protein were extracted from ovaries to detect scf/c-kit mRNA and protein level by real time PCR and Western-blot. Results Compared to control group,mRNA levels of DNMT1,DNMT3αand DNMT3βall significantly increased in ovaries exposed to 0.5μmol/L and 10μmol/L CdCl2（F=13.579,12.926,P=0.009,0.012）,while decreased in 50μmol/L CdCl2 group（F=10.620,P=0.031）.Meanwhile,DNMT1 and DNMT3αprotein levels were different among each group,which increased in10μmol/L group and decreased in other Cd-exposed groups（F=56.311,28.533,both P〈0.01）.The protein level of DNMT3βamong each group was not significantly different（F=2.047,P=0.186）. Conclusions The changes of DNA methytrasferase may be closely related to cadmium toxic effect on rat ovary.

关 键 词：镉 DNA甲基转移酶 卵巢 

分 类 号：R114[医药卫生—卫生毒理学]