色谱

色谱 ›› 2017, Vol. 35 ›› Issue (1): 1-7.DOI: 10.3724/SP.J.1123.2016.09001

• 研究论文 • 上一篇    下一篇

高效液相色谱-电喷雾-串联质谱研究四溴双酚A双(2-羟乙基醚)诱导大鼠嗜铬细胞瘤细胞呼吸链氧化磷酸化和能量代谢紊乱

刘倩, 胡立刚, 周群芳, 江桂斌   

  1. 中国科学院生态环境研究中心环境化学与生态毒理学国家重点实验室, 北京 100085
  • 收稿日期:2016-08-31 出版日期:2017-01-08 发布日期:2013-05-06
  • 通讯作者: 胡立刚,Tel:(010)62849129,E-mail:lghu@rcees.ac.cn.
  • 基金资助:

    国家重点基础研究发展计划（“973”）项目（2015CB453102）；国家自然科学基金-重点国际（地区）合作研究项目（21461142001）；中国科学院先导专项B（14040302）.

Tetrabromobisphenol A bis(2-hydroxyethyl ether) induced dysfunction of the respiratory chain oxidative phosphorylation and energy metabolism in rat pheochromocytoma cells by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry

LIU Qian, HU Ligang, ZHOU Qunfang, JIANG Guibin   

  1. State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
  • Received:2016-08-31 Online:2017-01-08 Published:2013-05-06
  • Supported by:

    National Basic Research Program of China (No. 2015CB453102); National Natural Science Foundation of China-Major International (Regional) Joint Project (No. 21461142001); Strategic Priority Research Program of the Chinese Academy of Sciences (No. 14040302).

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摘要：

四溴双酚A衍生物的毒理学研究亟须开展。已有研究发现四溴双酚A双（2-羟乙基醚）（tetrabromobisphenol A bis（2-hydroxyethyl ether），TBBPA-BHEE）可诱导大鼠嗜铬细胞瘤细胞（PC12）活性氧（reactive oxygen species，ROS）的生成。然而TBBPA-BHEE对PC12细胞线粒体呼吸链氧化磷酸化过程的干扰机制尚不明确，TBBPA-BHEE是否通过破坏线粒体功能干扰细胞能量代谢亟待进一步探讨。建立了基于HPLC-ESI-MS/MS分析PC12细胞内ATP、ADP、AMP及cAMP（cyclic AMP）浓度的方法，在此基础上评价了TBBPA-BHEE暴露对PC12线粒体呼吸链氧化磷酸化过程及能量代谢的影响。研究发现，TBBPA-BHEE可加速PC12线粒体呼吸链氧化磷酸化过程；TBBPA-BHEE诱导的PC12线粒体功能紊乱可引起细胞能量代谢紊乱。一方面揭示了TBBPA-BHEE对PC12潜在的毒性作用机制，另一方面也证实HPLC-ESI-MS/MS是研究细胞线粒体呼吸链氧化磷酸化及能量代谢过程的有力工具。

关键词: 大鼠嗜铬细胞瘤细胞, 高效液相色谱-电喷雾-串联质谱, 呼吸链氧化磷酸化, 能量代谢, 四溴双酚A衍生物

Abstract:

Toxicological effects of tetrabromobisphenol A (TBBPA) derivatives are highly concerned due to their increasing usages in diverse fields. It has been reported that tetrabromobisphenol A bis(2-hydroxyethyl ether) (TBBPA-BHEE), a representative of TBBPA derivatives, can induce cellular toxicity on rat pheochromocytoma cells (PC12) through reactive oxygen species mediated caspase activation. Nevertheless, underlying molecular mechanisms of intracellular reactive oxygen species (ROS) formation and effects of TBBPA-BHEE on PC12 energy metabolism remain unclear. This study developed the method for quantification of ATP, ADP, AMP and cAMP (cyclic AMP) simultaneously in PC12 with HPLC-ESI-MS/MS. The disturbance of TBBPA-BHEE on the respiratory chain oxidative phosphorylation and energy metabolism in PC12 cells was investigated via evaluation of the four compounds. It was shown that the ratio of ADP to ATP (ADP/ATP) decreased in PC12 cells after TBBPA-BHEE stimulation, implying that TBBPA-BHEE accelerated the process of respiratory chain oxidative phosphorylation in PC12 mitochondria. In addition, the concentrations of cAMP and the ratio of AMP to ATP (AMP/ATP) decreased in PC12 cells upon TBBPA-BHEE treatment, which suggested that TBBPA-BHEE potentially induced the mitochondrial dysfunction and led to the consequent energy metabolism imbalance in PC12. The results not only provided insight in the toxicological effects of TBBPA-BHEE, but also confirmed that HPLC-ESI-MS/MS was a powerful tool for studies on the cellular respiratory chain oxidative phosphorylation and energy metabolism.

Key words: energy metabolism, high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS), rat pheochromocytoma cells (PC12), respiratory chain oxidative phosphorylation, tetrabromobisphenol A (TBBPA) derivatives

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