Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/355877
Full metadata record
DC FieldValueLanguage
dc.contributor.CRUESPUNIVERSIDADE ESTADUAL DE CAMPINASpt_BR
dc.contributor.authorunicampCampos, Richard Piffer Soares de-
dc.contributor.authorunicampSilva, José Alberto Fracassi da-
dc.typeArtigopt_BR
dc.titleIndirect detection of superoxide in RAW 264.7 macrophage cells using microchip electrophoresis coupled to laser-induced fluorescencept_BR
dc.contributor.authorde Campos, Richard P. S.-
dc.contributor.authorSiegel, Joseph M.-
dc.contributor.authorFresta, Claudia G.-
dc.contributor.authorCaruso, Giuseppe-
dc.contributor.authorda Silva, Jose A. F.-
dc.contributor.authorLunte, Susan M.-
dc.subjectSuperóxidospt_BR
dc.subject.otherlanguageSuperoxidespt_BR
dc.description.abstractSuperoxide, a naturally produced reactive oxygen species (ROS) in the human body, is involved in many pathological and physiological signaling processes. However, if superoxide formation is left unregulated, overproduction can lead to oxidative damage to important biomolecules, such as DNA, lipids, and proteins. Superoxide can also lead to the formation of peroxynitrite, an extremely hazardous substance, through its reaction with endogenously produced nitric oxide. Despite its importance, quantitative information regarding superoxide production is difficult to obtain due to its high reactivity and low concentrations in vivo. MitoHE, a fluorescent probe that specifically reacts with superoxide, was used in conjunction with microchip electrophoresis (ME) and laser-induced fluorescence (LIF) detection to investigate changes in superoxide production by RAW 264.7 macrophage cells following stimulation with phorbol 12-myristate 13-acetate (PMA). Stimulation was performed in the presence and absence of the superoxide dismutase (SOD) inhibitors, diethyldithiocarbamate (DDC) and 2-metoxyestradiol (2-ME). The addition of these inhibitors resulted in an increase in the amount of superoxide specific product (2-OH-MitoE(+)) from 0.08 +/- 0.01 fmol (0.17 +/- 0.03 mM) in native cells to 1.26 +/- 0.06 fmol (2.5 +/- 0.1 mM) after PMA treatment. This corresponds to an approximately 15-fold increase in intracellular concentration per cell. Furthermore, the addition of 3-morpholino-sydnonimine (SIN-1) to the cells during incubation resulted in the production of 0.061 +/- 0.006 fmol (0.12 +/- 0.01 mM) of 2-OH-MitoE(+) per cell on average. These results demonstrate that indirect superoxide detection coupled with the use of SOD inhibitors and a separation method is a viable method to discriminate the 2-OH-MitoE(+) signal from possible interferencespt_BR
dc.relation.ispartofAnalytical and bioanalytical chemistrypt_BR
dc.relation.ispartofabbreviationAnal. bioanal. chem.pt_BR
dc.publisher.cityHeidelbergpt_BR
dc.publisher.countryAlemanhapt_BR
dc.publisherSpringerpt_BR
dc.date.issued2015-
dc.date.monthofcirculationSep.pt_BR
dc.language.isoengpt_BR
dc.description.volume407pt_BR
dc.description.issuenumber23pt_BR
dc.description.firstpage7003pt_BR
dc.description.lastpage7012pt_BR
dc.rightsFechadopt_BR
dc.sourceWOSpt_BR
dc.identifier.issn1618-2642pt_BR
dc.identifier.eissn1618-2650pt_BR
dc.identifier.doi10.1007/s00216-015-8865-1pt_BR
dc.identifier.urlhttps://link.springer.com/article/10.1007%2Fs00216-015-8865-1pt_BR
dc.description.sponsorshipFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPpt_BR
dc.description.sponsorshipCONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQpt_BR
dc.description.sponsorshipCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESpt_BR
dc.description.sponsordocumentnumbersem informaçãopt_BR
dc.date.available2021-02-16T10:58:19Z-
dc.date.accessioned2021-02-16T10:58:19Z-
dc.description.provenanceSubmitted by Sanches Olivia (olivias@unicamp.br) on 2021-02-16T10:58:19Z No. of bitstreams: 0. Added 1 bitstream(s) on 2021-05-25T12:08:33Z : No. of bitstreams: 1 000360412100010.pdf: 1057737 bytes, checksum: 7a87b01945bfbba025f8b3c74b1b8cae (MD5)en
dc.description.provenanceMade available in DSpace on 2021-02-16T10:58:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2015en
dc.identifier.urihttp://repositorio.unicamp.br/jspui/handle/REPOSIP/355877-
dc.contributor.departmentsem informaçãopt_BR
dc.contributor.departmentDepartamento de Química Analíticapt_BR
dc.contributor.unidadeInstituto de Químicapt_BR
dc.subject.keywordBioanalyticalmethodspt_BR
dc.subject.keywordFluorescencept_BR
dc.subject.keywordMicrochip electrophoresispt_BR
dc.identifier.source000360412100010pt_BR
dc.creator.orcid0000-0002-8037-8163pt_BR
dc.creator.orcid0000-0001-9522-6181pt_BR
dc.type.formArtigo de pesquisapt_BR
Appears in Collections:IQ - Artigos e Outros Documentos

Files in This Item:
File Description SizeFormat 
000360412100010.pdf1.03 MBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.