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Article: Simultaneous and ultrarapid determination of reactive oxygen species and reduced glutathione in apoptotic leukemia cells by microchip electrophoresis

TitleSimultaneous and ultrarapid determination of reactive oxygen species and reduced glutathione in apoptotic leukemia cells by microchip electrophoresis
Authors
KeywordsAcute promyelocytic leukemia
Apoptosis
Microchip electrophoresis
Miniaturization
Reactive oxygen species
Reduced glutathione
Issue Date2005
PublisherWiley - V C H Verlag GmbH & Co KGaA.
Citation
Electrophoresis, 2005, v. 26 n. 6, p. 1155-1162 How to Cite?
AbstractA 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 (DHR-123) can be converted intracellularly by ROS to the fluorescent rhodamine-123 (Rh-123), 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 NB4 cells induced by arsenic trioxide (As2O3) at low concentration (1-2 μM). Buthionine sulfoximine (BSO), in combination with As2O3 enhanced the decrease of reduced GSH to a great extent. The combined treatment of As2O3 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. © 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Persistent Identifierhttp://hdl.handle.net/10722/167912
ISSN
2023 Impact Factor: 3.0
2023 SCImago Journal Rankings: 0.541
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorQin, Jen_US
dc.contributor.authorYe, Nen_US
dc.contributor.authorYu, Len_US
dc.contributor.authorLiu, Den_US
dc.contributor.authorFung, Yen_US
dc.contributor.authorWang, Wen_US
dc.contributor.authorMa, Xen_US
dc.contributor.authorLin, Ben_US
dc.date.accessioned2012-10-08T03:12:51Z-
dc.date.available2012-10-08T03:12:51Z-
dc.date.issued2005en_US
dc.identifier.citationElectrophoresis, 2005, v. 26 n. 6, p. 1155-1162en_US
dc.identifier.issn0173-0835en_US
dc.identifier.urihttp://hdl.handle.net/10722/167912-
dc.description.abstractA 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 (DHR-123) can be converted intracellularly by ROS to the fluorescent rhodamine-123 (Rh-123), 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 NB4 cells induced by arsenic trioxide (As2O3) at low concentration (1-2 μM). Buthionine sulfoximine (BSO), in combination with As2O3 enhanced the decrease of reduced GSH to a great extent. The combined treatment of As2O3 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. © 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.en_US
dc.languageengen_US
dc.publisherWiley - V C H Verlag GmbH & Co KGaA.-
dc.relation.ispartofElectrophoresisen_US
dc.subjectAcute promyelocytic leukemia-
dc.subjectApoptosis-
dc.subjectMicrochip electrophoresis-
dc.subjectMiniaturization-
dc.subjectReactive oxygen species-
dc.subjectReduced glutathione-
dc.subject.meshApoptosis - Drug Effects - Physiologyen_US
dc.subject.meshArsenicals - Therapeutic Useen_US
dc.subject.meshButhionine Sulfoximine - Therapeutic Useen_US
dc.subject.meshElectrophoresis, Microchip - Methodsen_US
dc.subject.meshFluorescent Dyesen_US
dc.subject.meshGlutathione - Analysisen_US
dc.subject.meshHumansen_US
dc.subject.meshLasersen_US
dc.subject.meshLeukemia, Promyelocytic, Acute - Drug Therapy - Metabolismen_US
dc.subject.meshMiniaturizationen_US
dc.subject.meshOxides - Therapeutic Useen_US
dc.subject.meshReactive Oxygen Species - Analysisen_US
dc.subject.meshReproducibility Of Resultsen_US
dc.subject.meshRhodaminesen_US
dc.subject.meshSignal Transduction - Physiologyen_US
dc.subject.meshSpectrometry, Fluorescenceen_US
dc.subject.meshTumor Cells, Cultureden_US
dc.titleSimultaneous and ultrarapid determination of reactive oxygen species and reduced glutathione in apoptotic leukemia cells by microchip electrophoresisen_US
dc.typeArticleen_US
dc.identifier.emailFung, Y:ysfung@hku.hken_US
dc.identifier.authorityFung, Y=rp00697en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1002/elps.200410136en_US
dc.identifier.pmid15706575-
dc.identifier.scopuseid_2-s2.0-17144391406en_US
dc.identifier.hkuros104851-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-17144391406&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume26en_US
dc.identifier.issue6en_US
dc.identifier.spage1155en_US
dc.identifier.epage1162en_US
dc.identifier.isiWOS:000228170600015-
dc.publisher.placeGermanyen_US
dc.identifier.scopusauthoridQin, J=7402895572en_US
dc.identifier.scopusauthoridYe, N=8880430900en_US
dc.identifier.scopusauthoridYu, L=7404163512en_US
dc.identifier.scopusauthoridLiu, D=7410099966en_US
dc.identifier.scopusauthoridFung, Y=13309754700en_US
dc.identifier.scopusauthoridWang, W=36071339100en_US
dc.identifier.scopusauthoridMa, X=7404550382en_US
dc.identifier.scopusauthoridLin, B=34668182800en_US
dc.identifier.issnl0173-0835-

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