镉对体外培养大鼠肝细胞DNA甲基化的影响  

Effects of cadmium on DNA methylation in primary rat hepatocytes cultured in vitro

作　　者：朱家桥[1] 王玲玲[1] 程来洋 王棋文[1] 王亨[1] 刘学忠[1] 刘宗平[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《中国兽医学报》2014年第5期781-786,共6页Chinese Journal of Veterinary Science

基　　金：江苏省自然学基科金资助项目(BK2012264);江苏高校优势学科建设工程资助项目;扬州大学创新基金资助(027389604035349)

摘　　要：选用体外培养原代大鼠肝细胞为模型,用醋酸镉进行处理,用免疫荧光法检测了基因组DNA甲基化水平,用偏重亚硫酸氢盐测序法检测了MT、p53和Line1基因的DNA甲基化水平,用qRT-PCR检测了DNMTs基因的mRNA水平。结果显示,原代大鼠肝细胞在体外培养过程中基因组DNA甲基化水平具有缓慢下降的趋势,镉处理加速了这种下降的趋势;但p53、MT和Line1基因的DNA甲基化均未受镉的影响;维持DNA甲基化稳定的DNMTs表达水平受镉的影响呈现下降的趋势。表明镉对肝细胞的损害作用可能是通过降低DNMTs的表达水平,进而破坏基因组DNA甲基化的稳定来完成的;而镉对MT表达水平的影响似乎并非通过DNA甲基化途径来完成,因为在镉处理之前MT的DNA甲基化就已经处在较低的水平。Cadmium treated primary rat hepatocytes cultrued in vitro as the model. DNA methyal- tion of MT,p53 ,Linel and genome were examined by bisulfite sequencing （BSP） and immunoflu- orescence staining, respectively. Real-time RT-PCR were used to examine the transcription level of DNMTs. The results showed that genome DNA methylation was slowly decreased during in vitro culture of primary rat hepatocytes, and cadmium treatment accelerated this process. However, cad- mium treatment did not affect DNA mehtylaiton of p53. MT and Line1. The mRNA of DNMTs that maintain the genome DNA methyaltion decreased after cadmium treatment. These results indicate that cadmium may induce the disrupt genome DNA methyaliton to damage the hepatocytes. DNA methylaiton may not be an effective approach for cadmium to affect MT mRNA expression, because MT is hypomehtylaiton before cadmiun treatment.

关 键 词：镉 DNA甲基化 大鼠 原代肝细胞 

分 类 号：S859.8[农业科学—临床兽医学]