RESUMO
BACKGROUND: Recently, a prominent antiviral and macrophage stimulatory activity of cacao lignin-carbohydrate complex (LCC) has been reported. However, the solubility and sterility of LCC have not been considered yet. In the present study, complete solubilisation and sterilisation was achieved by autoclaving under mild alkaline conditions and the previously reported biological activities were re-examined. MATERIALS AND METHODS: LCCs were obtained by 1% NaOH extraction and acid precipitation, and a repeated extraction-precipitation cycle. Nitric oxide (NO) and cytokine productions were assayed by the Griess method and ELISA, respectively. Inducible NO synthase (iNOS) expression was determined by Western blot analysis. Superoxide anion, hydroxyl radical and nitric oxide radical-scavenging activity was determined by ESR spectroscopy. RESULTS: Cacao mass LCC showed reproducibly higher anti-HIV activity than cacao husk LCC. Cacao mass LCC, up to 62.5 µg/ml, did not stimulate mouse macrophage-like cells (RAW264.7 and J774.1) to produce NO, nor did it induce iNOS protein, in contrast to lipopolysaccharide (LPS). Cacao mass LCC and LPS synergistically stimulated iNOS protein expression, suggesting a different point of action. Cacao mass LCC induced tumour necrosis factor-α production markedly less than LPS, and did not induce interleukin-1ß, interferon-α or interferon-γ. ESR spectroscopy showed that cacao mass LCC, but not LPS, scavenged NO produced from NOC-7. CONCLUSION: This study demonstrated several new biological activities of LCCs distinct from LPS and further confirmed the promising antiviral and immunomodulating activities of LCCs.
Assuntos
Fármacos Anti-HIV/farmacologia , Cacau/química , Carboidratos/farmacologia , Fatores Imunológicos/farmacologia , Lignina/farmacologia , Extratos Vegetais/farmacologia , Animais , Western Blotting , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Espectroscopia de Ressonância de Spin Eletrônica , Ensaio de Imunoadsorção Enzimática , HIV-1/efeitos dos fármacos , HIV-1/crescimento & desenvolvimento , Humanos , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/virologia , Superóxidos/metabolismoRESUMO
Cacao husk lignin fractions, prepared by acid precipitation and 50% ethanol precipitation showed unexpectedly higher anti-human immunodeficiency virus (HIV) activity, as compared with the corresponding fractions from the cacao mass, amounting to the level comparable with that of popular anti-HIV compounds. The cacao husk lignin fractions also showed anti-influenza virus activity, but did not show antibacterial activity. The cacao husk lignin fractions synergistically enhanced the superoxide anion and hydroxyl radical scavenging activity of vitamin C. The cacao husk lignin fractions stimulated nitric oxide generation by mouse macrophage-like cells, to a level higher than that attained by lipopolysaccharide (LPS). The present study suggests the functionality of cacao husk lignin fractions as complementary alternative medicine.
