重庆市主城区长江和嘉陵江水中有机污染物对H4IIE细胞CYP1A1表达的诱导  

Induction of CYP1A1 expression of H4IIE cell after treated with the water organic pollutants from the Yangtze River and Jialing River

作　　者：崔志鸿[1] 李鹏[1] 刘晋祎[1] 刘胜学[1] 周燕虹[1] 曹佳[1] 

机构地区：[1]第三军医大学军事毒理学教研室,重庆400038

出　　处：《卫生研究》2008年第5期540-542,共3页Journal of Hygiene Research

基　　金：国家科技部西部引导项目(No.2003BA869C);国家博士后研究基金资助(No.20060390172)

摘　　要：目的以重庆市主城区大溪沟(嘉陵江)和寸滩(长江)两个点为代表,研究2004～2005年度重庆市水源水中有机污染物对H4IIE细胞细胞色素P4501A1(CYP1A1)表达的诱导。方法固相萃取法萃取水中有机污染物,将萃取的有机污染物溶于二甲基亚砜(DMSO)中,对H4IIE细胞进行染毒,用逆转录-聚合酶链式反应(RT-PCR)扩增CYP1A1mRNA,比较不同水样诱导H4IIE细胞CYP1A1mRNA表达的差异,用电泳迁移率实验(EMSA)检测CYP1A1启动子的激活情况。结果CYP1A1引物则可以将4个水样12h和24h处理组细胞的cDNA扩增出条带,48h处理组只有2005年1月寸滩样和2005年1月大溪沟样处理的细胞能扩增出CYP1A1基因的条带,所有样品的72h处理组细胞都没有CYP1A1基因的扩增;4个水样处理H4IIE细胞24h后,均可在细胞核内形成可与标记的XRE探针结合的芳烃受体-芳烃受体核转移蛋白(AHR-ARNT)二聚体,从而在AHR-ARNT二聚体蛋白处呈现发光条带。结论各水样均可激活CYP1A1基因的启动子XRE,也均可诱导H4IIE细胞CYP1A1mRNA的表达,CYP1A1mRNA的表达量随处理时间的差异有所不同。Objective To take Cuntan（Yangtze River） and Daxigou（Jialing River） as the representative,in order to study the cytochrome P450 1A1（CYP1A1） gene expression in H4IIE cell treated with the organic pollutants in Chongqing source water between 2004 and 2005.Methods The organic pollutants were extracted with solid phase extraction,dissolved the organic pollutants in dimethyl sulphoxide（DMSO）.The H4IIE cell was treated with the extracts.The differences of CYP1A1 mRNA expression among different treatment groups were detected by reverse transcriptase-polymerase chain reaction（RT-PCR）.The activities of CYP1A1 promotor were deteceted by electrophoretic mobility shift assay（EMSA）.Results CYP1A1 mRNA expressions in H4IIE cells were observed in all the groups at the time of 12h and 24h,as well as in the 2005.01 Daxigou and 2005.01 Cuntan treated groups at the time of 48h,but were not observed in all the groups at the time of 72h.The aryl hydrocarbon receptor-aryl hydrocarbon receptor nuclear translocator（AhR-ARNT） heterdimers were observed in all groups at the time of 24h.Conclusion All samples could induce the activity of the CYP1A1 gene promoter XRE,and could induce the expression of CYP1A1 mRNA,but the quantities of CYP1A1 mRNA expression were different according to the different treating time.

关 键 词：有机污染物 水污染 H4IIE细胞 细胞色素P450 1A1 

分 类 号：X522.032[环境科学与工程—环境工程] R994.6[医药卫生—毒理学]