双酚A对原代培养胎鼠脑多巴胺神经元的氧化损伤作用研究  

Study on bisphenol A induced primary cultured mesencephalic neuronal cell injury by oxidative stress

作　　者：林勇[1] 曾祥贵[1] 吴德生[1] 王霞[2] 屈卫东[2] 

机构地区：[1]四川大学华西公共卫生学院环境卫生学教研室,成都610041 [2]复旦大学公共卫生学院环境卫生学教研室

出　　处：《卫生研究》2006年第4期419-423,共5页Journal of Hygiene Research

基　　金：国家自然科学基金重点项目资助(No.30030120);国家高科技研究发展计划(863)基金专项经费(No.8632002AA601130)

摘　　要：目的 探讨双酚A对原代培养的胎鼠脑多巴胺神经元的毒性作用及损伤机制。方法 孕14～15天SD胎鼠中脑多巴胺神经元在无血清条件下分别培养4或7天后，分别加入1、10、25、50、100μmol／L双酚A，于24h和48h后检测细胞内活性氧（ROS）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、丙二醛（MDA）；流式细胞术检测细胞凋亡；酪氨酸羟化酶（TH）免疫组化用于鉴定和计数多巴胺神经元比例的变化。结果 与对照组相比，各处理组均可导致细胞内活性氧升高，双酚A≥50μmol／L时，可无选择地使细胞内活性1氧明显升高，同时使SOD、GSH水平明显降低，MDA水平明显升高；双酚A浓度≥50μmol／L时可使凋亡细胞数显著增加。酪氨酸羟化酶阳性细胞数在25μmol／L以上时随双酚A浓度增加其比例显著降低。结论 双酚A可通过氧化损伤导致体外培养的多巴胺神经元凋亡。Objective To investigate the effect and mechanism of bisphenol A, a common EDCs, on the primary cultured mesencephalic neuronal cell injury. Methods mesencephalic neuronal cells were obtained from embryonic 14- 15day SD fetuses and were cultured in serum free medium for 4 or 7 days in vitro, 1,10,25,50 and 100μmol/L bisphenol A were added to the medium and intracellular ROS, SOD, MDA, GSH were measured after 24 or 48hours, respectively. Flow cytometry was used to detect the neuronal cell apoptosis. Tyrosine hydroxylase immunohistochemistry was used to identity and count the ratio of dopaminergic neuronal cell. Results The intracellular ROS was dose dependently increased by bisphenol A in all treated groups, and it appeared to be a nonselective effect when the concentration of bisphenol A were higher than 50μmol/L, and concomitantly decreased SOD, GSH and increased MDA. The apoptotic neuronal cells were significantly increased at 50μmol/L and 100μmol/L. The ratio of Tyrosine hydroxylase positive neuronal cell was dose dependently decreased with the concentration of bisphenol A. Conclusion ROS induced by bisphenol A may play a major role in its cytotoxicity on dopaminergic neuronal cell.

关 键 词：双酚A 无血清原代培养 多巴胺神经元 活性氧 凋亡 

分 类 号：R994.6[医药卫生—毒理学] Q426[医药卫生—药学]