Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line.
Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line.
复制标题
DOI：
10.1016/j.taap.2011.03.009
复制
发表时间：
2011-05-15
影响因子：
3.8
通讯作者：
Burghardt RC
中科院分区：
文献类型：
--
作者：
Barhoumi R;Mouneimne Y;Ramos E;Morisseau C;Hammock BD;Safe S;Parrish AR;Burghardt RC
实时动态分析多环芳烃(PAHs)及其代谢产物在活细胞内的摄取和代谢，可能为研究PAHs引起细胞损伤的遗传毒性和非遗传毒性机制提供新的见解。本研究结合使用多光子光谱分析代谢物标准产生的代谢物光谱和“高级分解过程”，在24小时内识别和量化了存活培养的大鼠肝细胞中最重要的多环芳烃之一(苯并[a]芘，BaP)的摄取、分配和代谢物形成。应用先进的拆分工艺，可同时鉴定活细胞中的8种代谢物。使用特定的微粒体环氧化物水解酶抑制剂、葡萄糖醛酸化和硫酸盐化抑制剂以及几种代谢物标准混合物验证了这种混合过程的准确性。结果表明，双光子显微成像技术优于传统的荧光成像技术，分离过程是一种适用于分析活细胞中BaP代谢物的数学技术，尤其适用于分析最终致癌物质苯并[a]芘-r-7，t-8-二氢二醇-t-9，10-环氧化物的变化。因此，双光子获取和分解过程的结合将为BaP和其他多环芳烃改变细胞内稳态的细胞和分子机制提供重要的见解。
Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an “advanced unmixing process”, identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 hr. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.
登录
查看更多内容
影响因子：
4.7
作者：
GMUR, DJ;VARANASI, U
通讯作者：
VARANASI, U
影响因子：
3.5
作者：
Barhoumi, Rola;Awooda, Igbal;Burghardt, Robert C.
通讯作者：
Burghardt, Robert C.
影响因子：
--
作者：
Ramesh, A;Inyang, F;Knuckles, ME
通讯作者：
Knuckles, ME
影响因子：
3.8
作者：
Barhoumi, R;Mouneimne, Y;Burghardt, RC
通讯作者：
Burghardt, RC
DOI：
10.1080/009841096161203
发表时间：
1996-08-09
期刊：
JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH
影响因子：
--
作者：
Miles, PR;Ma, JYC;Miller, MR
通讯作者：
Miller, MR