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题名: Simultaneous and ultrarapid determination of reactive oxygen species and reduced glutathione in apoptotic leukemia cells by microchip electrophoresis
作者: Qin, JH;  Ye, NN;  Yu, LF;  Liu, DY;  Fung, YS;  Wang, W;  Ma, XJ;  Lin, BC
通讯作者: 林炳承
关键词: acute promyelocytic leukemia ;  apoptosis ;  microchip electrophoresis ;  miniaturization ;  reactive oxygen species ;  reduced glutathione
刊名: ELECTROPHORESIS
发表日期: 2005-03-01
DOI: 10.1002/elps.200410136
卷: 26, 期:6, 页:1155-1162
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1807,1802
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: A microchip electrophoresis method coupled with laser-induced fluorescence (LIF) detection was established for simultaneous determination of two kinds of intracellular signaling molecules (reactive oxygen species, ROS, and reduced glutathione, GSH) related to apoptosis and oxidative stress. As the probe dihydrorhodamine-123 (DHR123) can be converted intracellularly by ROS to the fluorescent rhodamine-123 (Rh123), and the probe naphthalene-2,3-dicarboxaldehyde (NDA) can react quickly with GSH to produce a fluorescent adduct, rapid determination of Rh-123 and GSH was achieved on a glass microchip within 27 s using a 20 mm borate buffer (pH 9.2). The established method was tested to measure the intracellular ROS and GSH levels in acute promyelocytic leukemia (APL)-derived NB4 cells. An elevation of intracellular ROS and depletion of GSH were observed in apoptotic N134 cells induced by arsenic trioxide (AS(2)O(3)) at low concentration (1-2 mu m). Buthionine sulfoximine (BSO), in combination with AS(2)O(3) enhanced the decrease of reduced GSH to a great extent. The combined treatment of AS(2)O(3) and hydrogen peroxide (H2O2) led to an inverse relationship between the concentrations of ROS and GSH obtained, showing the proposed method can readily evaluate the generation of ROS, which occurs simultaneously with the consumption of the inherent antioxidant.
关键词[WOS]: CAPILLARY-ELECTROPHORESIS ;  ARSENIC TRIOXIDE ;  SYSTEM ;  DERIVATIZATION ;  CHROMATOGRAPHY ;  QUANTITATION ;  LINES
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000228170600015
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/92275
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Univ Hong Kong, Dept Chem, Hong Kong, Hong Kong, Peoples R China

Recommended Citation:
Qin, JH,Ye, NN,Yu, LF,et al. Simultaneous and ultrarapid determination of reactive oxygen species and reduced glutathione in apoptotic leukemia cells by microchip electrophoresis[J]. ELECTROPHORESIS,2005,26(6):1155-1162.
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