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采用代谢流量分析方法评估二噁英对细胞的代谢干扰

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张保琴, 张海军, 王龙星, 王金成, 杨常青, 陈吉平. 采用代谢流量分析方法评估二噁英对细胞的代谢干扰[J]. 环境化学, 2012, 31(11): 1797-1802.
引用本文: 张保琴, 张海军, 王龙星, 王金成, 杨常青, 陈吉平. 采用代谢流量分析方法评估二噁英对细胞的代谢干扰[J]. 环境化学, 2012, 31(11): 1797-1802.
shu
ZHANG Baoqin, ZHANG Haijun, WANG Longxing, WANG Jincheng, YANG Changqing, CHEN Jiping. The influence of dioxin on cell metabolism evaluated by the metabolic network flexibility analysis[J]. Environmental Chemistry, 2012, 31(11): 1797-1802.
Citation: ZHANG Baoqin, ZHANG Haijun, WANG Longxing, WANG Jincheng, YANG Changqing, CHEN Jiping. The influence of dioxin on cell metabolism evaluated by the metabolic network flexibility analysis[J]. Environmental Chemistry, 2012, 31(11): 1797-1802.
shu

采用代谢流量分析方法评估二噁英对细胞的代谢干扰

  • 1.

    中国科学院大连化学物理研究所, 大连, 116023;

  • 2.

    满洲里出入境检验检疫局技术中心, 满洲里, 021400

  • 基金项目:

    国家重点基础研究发展计划(2009CB421602)资助.

The influence of dioxin on cell metabolism evaluated by the metabolic network flexibility analysis

  • 1.

    Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China;

  • 2.

    Manzhouli Entry-Exit Inspection and Quarantine Bureau, Manzhouli, 021400, China

  • Fund Project:

  • 摘要: 以肝癌细胞HepG2为供试细胞,选用5个不同浓度的二噁英(0、0.001、0.01、0.1和1.0 nmol·L-1),探讨2,3,7,8-TCDD对HepG2细胞的毒性效应.采用90%甲醇离心法实现蛋白与代谢产物的分离,建立了高效液相色谱-串联三重四极杆质谱(HPLC-MS/MS)分析胞外液中的23种代谢产物(20种氨基酸、乳酸、甘油和尿素)的方法.建立的HPLC-MS/MS分析方法对各化合物的加标回收率可达91%-105%之间,化合物浓度的标准偏差均小于10%.在此基础上,结合代谢流量分析系统研究了2,3,7,8-TCDD对HepG2细胞内代谢流的影响.结果表明,随着浓度的增高,2,3,7,8-TCDD对HepG2细胞代谢的干扰作用呈增强趋势.2,3,7,8-TCDD降低了细胞对葡萄糖的吸收,并进一步抑制了HepG2细胞糖酵解的速率.此外,2,3,7,8-TCDD导致了HepG2细胞乳酸代谢流的明显增加,由此使细胞产生了过量的乳酸.
    Abstract: This study was initiated to explore the toxic effects of dioxins in HepG2 cells when exposed to 2,3,7,8-TCDD at five different concentrations (0, 0.001, 0.01, 0.1, 1.0 nmol稬-1). The separation of proteins and metabolites was achieved by the centrifugation method with 90% methanol as solvent, and then 23 extracellular metabolites (20 underivatized amino acids, lactate, glycerol and urea) were qualitatively and quantitatively analyzed by HPLC-MS/MS. The recoveries of the 23 cell metabolites were 91%-105% and the relative standard deviations (RSDs) of peak areas of the extracted ion chromatograms were below 10%. Meanwhile, the influence of 2,3,7,8-TCDD on the metabolic flux analysis in HepG2 cells was evaluated. The results showed that the disturbance of 2,3,7,8-TCDD on the metabolism of HepG2 increased its concentration. The addition of 2,3,7,8-TCDD decreased the cell absorption of glucose and further inhibited the glycolysis. On the other hand, exposure to 2,3,7,8-TCDD induced a significant increase of lactic acid metabolic flux, and thus caused excessive lactic acid in HepG2 cells.
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    [2] Paul R W, April M W, Alan M S, et al. Predicting human developmental toxicity of pharmaceuticals using human embryonic stem cells and metabolomics[J]. Toxicol Appl Pharm, 2010, 247: 18-27
    [3] Gabriela G C, Jessica A Q, Alan M S, et al. Identification of small molecules from human embryonic stem gells using metabolomics[J]. Stem Cells Dev, 2007, 16: 869-882
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  • 收稿日期:  2012-01-10
张保琴, 张海军, 王龙星, 王金成, 杨常青, 陈吉平. 采用代谢流量分析方法评估二噁英对细胞的代谢干扰[J]. 环境化学, 2012, 31(11): 1797-1802.
引用本文: 张保琴, 张海军, 王龙星, 王金成, 杨常青, 陈吉平. 采用代谢流量分析方法评估二噁英对细胞的代谢干扰[J]. 环境化学, 2012, 31(11): 1797-1802.
shu
ZHANG Baoqin, ZHANG Haijun, WANG Longxing, WANG Jincheng, YANG Changqing, CHEN Jiping. The influence of dioxin on cell metabolism evaluated by the metabolic network flexibility analysis[J]. Environmental Chemistry, 2012, 31(11): 1797-1802.
Citation: ZHANG Baoqin, ZHANG Haijun, WANG Longxing, WANG Jincheng, YANG Changqing, CHEN Jiping. The influence of dioxin on cell metabolism evaluated by the metabolic network flexibility analysis[J]. Environmental Chemistry, 2012, 31(11): 1797-1802.
shu

采用代谢流量分析方法评估二噁英对细胞的代谢干扰

  • 1.  中国科学院大连化学物理研究所, 大连, 116023;
  • 2.  满洲里出入境检验检疫局技术中心, 满洲里, 021400
  • 收稿日期:  2012-01-10
基金项目:

国家重点基础研究发展计划(2009CB421602)资助.

摘要: 以肝癌细胞HepG2为供试细胞,选用5个不同浓度的二噁英(0、0.001、0.01、0.1和1.0 nmol·L-1),探讨2,3,7,8-TCDD对HepG2细胞的毒性效应.采用90%甲醇离心法实现蛋白与代谢产物的分离,建立了高效液相色谱-串联三重四极杆质谱(HPLC-MS/MS)分析胞外液中的23种代谢产物(20种氨基酸、乳酸、甘油和尿素)的方法.建立的HPLC-MS/MS分析方法对各化合物的加标回收率可达91%-105%之间,化合物浓度的标准偏差均小于10%.在此基础上,结合代谢流量分析系统研究了2,3,7,8-TCDD对HepG2细胞内代谢流的影响.结果表明,随着浓度的增高,2,3,7,8-TCDD对HepG2细胞代谢的干扰作用呈增强趋势.2,3,7,8-TCDD降低了细胞对葡萄糖的吸收,并进一步抑制了HepG2细胞糖酵解的速率.此外,2,3,7,8-TCDD导致了HepG2细胞乳酸代谢流的明显增加,由此使细胞产生了过量的乳酸.

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