RESUMO
According to the reported gene sequence of Rhizopus oryzae glucoamylases, the glucoamylase gene containing four introns was cloned from the total DNA of the natural Rhizopus arrhizu. Specific primers were designed to delete introns by overlapping PCR and a new cDNA sequence of Rhizopus arrhizu glucoamylase was obtained. The accession number in gene bank is DQ903853. This gene is successfully expressed in the Picha pastoris, producing a new protein with a high activity of glucoamylase.
Assuntos
Biocatálise , Western Blotting , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Proteínas Fúngicas , Genética , Metabolismo , Regulação Enzimológica da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Glucana 1,4-alfa-Glucosidase , Genética , Metabolismo , Dados de Sequência Molecular , Pichia , Genética , Proteínas Recombinantes , Metabolismo , Rhizopus , Genética , Análise de Sequência de DNARESUMO
<p><b>OBJECTIVE</b>To study the effect and mechanism of sodium ferulate (SF) in preventing and treating ozone (O3) induced lung oxidative injury in mice.</p><p><b>METHODS</b>Lung oxidative injury model mice were established by making them inhale O3. The activity of anti-oxidase and membranous microviscosity in epithelial cells in the lung of mice were determined, and the ultrastructural change of lung tissues was observed with electromicroscopy.</p><p><b>RESULTS</b>Activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were reduced, while membranous lipo-microviscosity significantly increased in the pulmonary epithelial cells of model mice, revealing ultrastructural change. These abnormal changes were reversed by SF treatment, which was manifested as the significantly raised activities of SOD and GSH-Px after treatment with high and moderate doses of SF, showing a significant difference compared with those in the model group (P<0.01). Membranous lipo-microviscosity basically approached that in the control group (P>0.05); electron microscopic examination showed a basically normal morphological structure of pulmonary epithelial cells, with the change in lung injury significantly milder than that in the model group.</p><p><b>CONCLUSION</b>O3 could induce oxidative injury of lungs in mice, and SF could enhance the anti-oxidation capacity of mice and scavenge the oxygen free radicals so as to alleviate the injury.</p>
Assuntos
Animais , Masculino , Camundongos , Ácidos Cumáricos , Farmacologia , Usos Terapêuticos , Glutationa Peroxidase , Metabolismo , Pulmão , Pneumopatias , Tratamento Farmacológico , Lipídeos de Membrana , Metabolismo , Ozônio , Superóxido Dismutase , Metabolismo , ViscosidadeRESUMO
<p><b>OBJECTIVE</b>To study the effect of sodium ferulate (SF), an active component of Radix Angelica, on lung damage induced by ozone (03).</p><p><b>METHODS</b>Mice model of lung injury was induced by ozone inhalation and treated with SF. The level of lipid peroxide and microviscosity in alveolar epithelial cell membrane of the mice was determined, and the structural change of lung cells was observed by microscopy.</p><p><b>RESULTS</b>Ozone could increase the level of malondialdehyde (MDA) and the microviscosity in alveolar epithelial cell membrane, and induce inflammatory changes in morphologic structure. These abnormal changes were improved after SF administration, which was manifested as alleviation of heightened microviscosity, increase of membrane fluidity, as well as the basically normalized pulmonary cellular structure under microscope.</p><p><b>CONCLUSION</b>SF has a preventive effect against oxidized pulmonary injury induced by ozone, the action of which could be through scavenging oxygen free radicals, reducing lipid peroxide production, increasing membranous fluidity and mitigating inflammatory changes in cell structure. sodium ferulate, ozone, malondialdehyde, membranous fluidity, morphology</p>