2,2′,4,4′-四溴联苯醚(BDE-47)诱导孕烷X受体(PXR)活化能力的探讨  

The effect of BDE-47 on activation of pregnane X receptor(PXR)

作　　者：胡辛楠[1,2] 张建清[2] 田亚楠 雷毅雄[1] 方道奎[2] 黄海燕[2] 蒋友胜[2] 周健[2] 

机构地区：[1]广州医学院公共卫生与全科医学学院,广东广州510182 [2]深圳市疾病预防控制中心 [3]美国德克萨斯州A&M大学

出　　处：《毒理学杂志》2010年第1期10-15,共6页Journal of Toxicology

基　　金：国家自然科学基金项目(30771778)

摘　　要：目的探讨2,2′,4,4′-四溴联苯醚(BDE-47)是否为孕烷X受体(pregnane X receptor,PXR)的诱导剂及其诱导PXR受体下游基因细胞色素P4503A4(CYP3A4)的转录表达能力。方法采用CCK-8法测定分析BDE-47对人肝肿瘤细胞株HepG2的细胞毒性作用,并以BDE-47分别处理双萤光素酶hPXR报告基因系统和稳定高表达hPXR的HepG2细胞株,观察其对CYP3A4的诱导作用和对其mRNA及蛋白表达的诱导作用。结果BDE-47对HepG2细胞有明显的细胞毒性作用,且在6～48h呈明显剂量-时间-效应关系(P<0.01)。48h为毒作用兴奋点,其半数抑制浓度(IC50)为110μmol/L。BDE-47能诱导CYP3A4表达量的增高,呈明显剂量-时间-效应关系(P<0.01)。Q-PCR和Western Blot分析发现,其对CYP3A4的mRNA转录和蛋白表达有显著诱导作用,并呈剂量-效应关系(P<0.01),对PXR受体诱导能力明显高于已知的阳性诱导物利福平。结论在本试验条件下,BDE-47是PXR受体的强力诱导剂,可能通过激活PXR受体而发挥其一系列毒性作用。Objective To explore whether can Tetra-brominated diphenyl ethers（BDE-47） induce CYP3A4 by activating the Pregnane X receptor（PXR）.Methods After exposure to BDE-47,HepG2 cell toxic effect was analyzed by Cell Counting Kit-8（CCK-8） assay.The dual-luciferase reporter assay system and stable expression system with karyocyte HepG2 cell of hPXR gene were constructed,then the induction ability of BDE-47 on PXR and CYP3A4 expression in mRNA and protein level were investigated and analyzed.Results BDE-47 had significant cytotoxicity to HepG2 cells depending on time and dose,P0.01.IC50 of BDE47 to HepG2 was 110μmol/L and expressed the strongest toxicity effect at 48 hrs exposure.Luciferase determination results suggested that BDE-47 could induce the expression of CYP3A4 increase,and showed dose-time-effect relationship significantly.Q-PCR and Western Blot analysis showed that the mRNA and protein of CYP3A4 transcription expression could be significantly induced by BDE-47 in a dose-dependent mannerP0.01.Conclusion BDE-47 shows cytotoxicity toward HepG2 cells and can activate PXR,and induce the controlling gene CYP3A4 elevated.Toxicity effect of BDE-47 may be expressed by PXR activation.

关 键 词：2 2′ 4 4′-四溴联苯醚(BDE-47) PXR受体 CYP3A4 hPXR双萤光素酶报告基因 

分 类 号：R994.6[医药卫生—毒理学]