丙烯酰胺对F344大鼠肝毒性的分子机制研究  

Molecular mechanisms of hepatoxicity caused by acrylamide in F344 rats

作　　者：赖怡[1,2] 陈佳红[1,2] 张航[1,2] 乐聪[1,2] 姜岩[1,3] 陈涛[1,2] 

机构地区：[1]苏州大学医学部公共卫生学院,江苏苏州215123 [2]江苏省老年病预防与转化医学重点实验室,江苏苏州215123 [3]苏州大学医学部生物科学与基础医学学院,江苏苏州215123

出　　处：《癌变．畸变．突变》2016年第3期174-178,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：教育部留学回国人员科研启动基金;江苏高校优势学科建设工程资助项目;国家自然科学基金(81300143)

摘　　要：目的:通过检测丙烯酰胺对大鼠肝脏基因表达、DNA甲基化以及基因突变的影响,探讨其肝脏毒性的分子机制。方法:以含50μg/mL丙烯酰胺的饮用水染毒7周龄F344雌性大鼠28 d;采用荧光定量PCR方法检测肝脏细胞增殖以及DNA甲基化调控相关基因的mRNA表达水平;结合重亚硫酸盐的限制性内切酶分析方法检测Cdkn1a/Cdkn2a基因启动子区和重复序列Line-1的DNA甲基化水平;PCR扩增测序方法检测H-ras基因突变。结果:丙烯酰胺引起大鼠肝脏中Dnmt3a、Cdkn1a、Cdkn2a、Jun和Line-1的mRNA水平表达下降;未检测到Cdkn1a/Cdkn2a基因启动子区DNA和重复序列Line-1甲基化的变化;测序结果发现丙烯酰胺未引起H-ras基因突变。结论:丙烯酰胺可引起大鼠肝脏中细胞增殖相关基因表达的变化,但无DNA甲基化改变和基因突变。OBJECTIVE：This study was aimed to investigate molecular mechanisms of acrylamide（AA）-induced hepatotoxicity by examining the effects on mRNA expression,DNA methylation and gene mutations in F344 female rats.METHODS：F344 female rats aged 7 weeks were orally dosed with 0 and 50 μg/mL of AA in drinking water for 28 d.Using qPCR,we detected mRNA expression level of genes which were involved with cell proliferationand and with regulation of DNA methylation in livers.The promoter DNA methylation status of Cdknla,Cdkn2 a and repetitive sequence Line-1 were examined by COBRA.We also examined the mutation levels of H-ras gene by PCR sequencing.RESULTS：Compared with control,AA decreased mRNA expression levels of Dnmt3 a,Cdknla,Cdkn2 a,Jun and Line-1 in F344 rat liver.No significant change was found in the global DNA methylation and the promoter methylation status of Cdknla and Cdkn2 a.Moreover,no gene mutation of H-ras was found.CONCLUSION：AA induced expression changes of genes which were involved in DNA methylation regulation and cell proliferation but no alteration in DNA methylation in rat livers.However,no gene mutation was detected

关 键 词：丙烯酰胺 MRNA DNA甲基化 肝毒性 基因突变 

分 类 号：R114[医药卫生—卫生毒理学]