Microplate Superoxide Dismutase Assay Employing a Nonenzymatic Superoxide Generator

Ewing, James F.; Janero, David R.

doi:10.1006/abio.1995.0014

Cited by 392 publications

(204 citation statements)

References 4 publications

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“…Assim, dois radicais superóxido doam, cada um, um elétron para o NBT, reduzindo e formando a formazam, que é monitorado em um comprimento de onda de 560 nm, em espectrofotômetro UV-Vis modelo DTX 800, Beckman. 33 Foram preparados inicialmente os reagentes NBT (250 µM), NADH (390 µM) e PMS (10 µM) em tampão Tris-HCl (16 mM, pH=8,0) e, em seguida, em uma placa de 96 poços foram adicionados: 50 µL de solução dos extratos metanólicos de B. purpurea nas concentrações de 10 a 1000 µg/mL, 100 µL de NADH e 100 µL de NBT. A absorbância inicial foi obtida a partir da leitura da microplaca a 560 nm.…”

Section: Avaliação Do Potencial Antioxidante Frente O Radical Ounclassified

Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

Santos¹,

Almeida²,

Lima³

et al. 2014

Quím. Nova

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FLAVONOID PROFILES AND EVALUATION OF ANTIOXIDANT POTENTIAL AND CYTOTOXICITY OF AMAZONIANBauhinia purpurea (Fabaceae). Plants of the Bauhinia species (Fabaceae) are used in the treatment of several infections and diseases, especially diabetes. In this paper, with an aim to contribute to the knowledge of the chemistry and therapeutic potential of the Amazonian species of the genus Bauhinia, flavonoid profiles in the leaves and branches of Bauhinia purpurea collected in Manaus and São Gabriel da Cachoeira/AM were analyzed by HPLC-DAD and ESIMS. Differences in the flavonoid profiles were detected along with the presence of rutin and isoquercitrin in the leaves collected in Manaus. All samples showed antioxidant activity, however, no cytotoxic activity against cell lines MDA-MB435 (breast), , and SF-295 (glioblastoma) could be detected.Keywords: Bauhinia purpurea; rutin; isoquercitrin. INTRODUÇÃOO gênero Bauhinia pertencente à família Fabaceae (subfamília: Cercideae) contém cerca de 300 espécies que são conhecidas popularmente no Brasil como pata-de-vaca pelo formato bilobado de suas folhas. 1 Elas são usadas na medicina tradicional em diversas partes do mundo por suas propriedades analgésica, antidiabética, antiinflamatória, antimicrobiana, adstrigente, diurética no tratamento de inchaços e tumores estomacais, diarréia e úlcera. 2-5 O uso popular mais comumente relatado para esse gênero no Brasil está relacionado à atividade hipoglicemiante, com várias espécies usadas para esse fim, em especial Bauhinia forficata, que é amplamente comercializada em variadas formas farmacêuticas. 6,7 As espécies de Bauhinia contêm diversas substâncias, como flavonoides livres e glicosilados, terpenoides, esteroides, ácidos fenólicos e alcaloides. 6-9 De espécimes asiáticos de B. purpurea foram isolados além de flavonoides, como kaempferol-3-galactosídio, isoquercitrina, rutina e astragalina, compostos raros da classe oxepinos, bauhiniastatinas, alguns com atividade citotóxica. [10][11][12] De um modo geral, as propriedades terapêuticas das espécies, especialmente as atividades antioxidante e hipoglicemiante, têm sido correlacionadas à presença de compostos fenólicos, em especial de flavonoides glicosilados, como a kaempferitrina, vinculada à ação hipoglicemiante da espécie B. forficata. [13][14][15] O estudo da espécie B. purpurea frente à atividade antioxidante vem ganhando destaque no estudo do gênero, já que seus extratos têm exibido potencial intermediário ou equivalente aos padrões geralmente utilizados nos testes para esta atividade. As substâncias fenólicas, atribuídas como as principais responsáveis por essas atividades, tem sido investigadas por meios cromatográficos e espectrométricos, na busca para estabelecer um perfil cromatográfico dentro da espécie. [16][17][18][19] Na região amazônica, o uso de espécies desse gênero não é diferente, havendo relatos para o tratamento de diabetes nos estados do Acre, Amazonas e Pará. As espécies são encontradas em muitas feiras e comércios a céu aberto, especialmente folhas, para uso co...

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Section: Avaliação Do Potencial Antioxidante Frente O Radical Ounclassified

Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

Santos¹,

Almeida²,

Lima³

et al. 2014

Quím. Nova

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“…We investigated SOD activity using a biochemical assay (1 day post-MCAO; nϭ6 mice per group) (Ewing and Janero, 1995). Brains were homogenized in phosphate-buffered saline and frozen immediately in liquid nitrogen.…”

Section: Sod Activitymentioning

confidence: 99%

“…Brains were homogenized in phosphate-buffered saline and frozen immediately in liquid nitrogen. Aliquots of brain supernatants were processed as previously described (Ewing and Janero, 1995). Brain supernatants were added (25 l) to a 125 l solution containing 50 mM phosphate buffer (pH 7.4, 0.1 mM EDTA, 50 M NBT, 78 M NADH).…”

Section: Sod Activitymentioning

confidence: 99%

Decreased infarct size after focal cerebral ischemia in mice chronically infected with Toxoplasma gondii

et al. 2007

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Abstract-To determine whether Toxoplasma gondii infection could modify biological phenomena associated with brain ischemia, we investigated the effect of permanent middle cerebral artery occlusion (MCAO) on neuronal survival, inflammation and redox state in chronically infected mice. Infected animals showed a 40% to 50% decrease of infarct size compared with non-infected littermates 1, 4 and 14 days after MCAO. The resistance of infected mice may be associated with increased basal levels of anti-inflammatory cytokines and/or a marked reduction of the MCAO-related brain induction of two pro-inflammatory cytokines, tumor necrosis factor-alpha and interferon-gamma (IFN␥). In addition, potential anti-inflammatory/neuroprotective factors such as nerve growth factor, suppressor of cytokine signaling-3, superoxide dismutase activity, uncoupling protein-2 and glutathione (GSH) were upregulated in the brain of infected mice. Consistent with a role of GSH in central cytokine regulation, GSH depletion by diethyl maleate inhibited Toxoplasma gondii lesion resistance by increasing the proinflammatory cytokine IFN␥ brain levels. Overall, these findings indicate that chronic toxoplasmosis decisively influences both the inflammatory molecular events and outcome of cerebral ischemia.

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“…An indirect spectrophotometric method used for the assessment of SOD activity in erythrocytes and liver homogenate was based on the generation of O . 2 -by a mixture of nitro blue tetrazolium, NADH and phenazine methosulfate (Ewing and Janero, 1995). The glutathione peroxidase activity was assayed by a modification of the method of Tappel (1978).…”

Section: Clinical Chemistry and Oxidative Stress Parameters And Determentioning

confidence: 99%

Safety assessment of sanguiritrin, alkaloid fraction of Macleaya cordata, in rats

Psotová¹,

Večeřa²,

Zdařilová³

et al. 2006

Vet. Med.

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ABSTRACT:The subchronic safety of sanguiritrin, a mixture of sanguinarine (SA) and chelerythrine (CHE) quaternary benzo[c]phenanthridine alkaloids (QBA), obtained from Macleaya cordata was assessed. Rats were fed a diet containing 120 ppm sanguiritrin (100 ppm QBA) for 109 days. The feed consumption and the animal weight were monitored. The content of QBA in selected tissues and plasma was determined using HPLC. It was evidenced that 2% of QBA were absorbed through the GIT while 98% were excreted in the feces. In plasma, bilirubin, urea, creatinine, glomerular filtration, AST, ALT, GMT, ALP and total antioxidant capacity were determined. In liver, GSH level, lipoperoxidation products, SOD and GPx activities and total amount of cytochrome P450 were evaluated. Damage to nuclear DNA was assessed; a 32 P-postlabeling assay proved that no DNA-adducts were detected in nuclear and mitochrondrial DNA in liver. No adverse effects were observed on rat organism. QBA had no influence on the gut mucosal epithelium, liver tissue and any biochemical parameters tested. Oxidative stress was not manifested during the experiment.

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Microplate Superoxide Dismutase Assay Employing a Nonenzymatic Superoxide Generator

Cited by 392 publications

References 4 publications

Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

Decreased infarct size after focal cerebral ischemia in mice chronically infected with Toxoplasma gondii

Safety assessment of sanguiritrin, alkaloid fraction of Macleaya cordata, in rats

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