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）（ tetrabromobisphe?nol 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。
Toxicological effects of tetrabromobisphenol A ( TBBPA) derivatives are highly con?cerned due to their increasing usages in diverse fields. It has been reported that tetrabromobis?phenol 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 intra?cellular 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.
参考文献
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