四角蛤蜊不同水提醇沉部位降血糖作用及多糖和蛋白成分研究
Study on the Anti-Hyperglycemic Activity and Polysaccharide and Protein Components in Different Alcohol Precipitate Fraction of Mactra Veneriformis
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摘要: 目的 筛选四角蛤蜊的降血糖活性部位，测定水提液的不同浓度醇沉部位的多糖和蛋白含量，并对蛋白质类成分进行初步鉴定。方法 针对四氧嘧啶诱导的糖尿病模型ICR小鼠通过剪尾取血测定其给药前后的血糖，摘眼球取血测定小鼠血清胰岛素、甘油三酯及总胆固醇的含量；采用蒽酮-硫酸法和BCA法测定多糖和蛋白质含量；利用SDS-PAGE凝胶电泳结合LC-MS/MS分析蛋白质类成分。结果 40%、60%和80%醇沉部位均可降低小鼠血糖且能降低小鼠血清胰岛素和甘油三酯；80%醇沉部位可降低小鼠血清胆固醇。四角蛤蜊20%、40%、60%、80%醇沉部位的多糖含量分别为21.45%、45.82%、14.61%、22.84%；水溶性蛋白的含量分别为23.56%、16.85%、28.60%、36.10%。水溶性蛋白质类成分主要包括肌球蛋白、原肌球蛋白、肌动蛋白、钙调蛋白、弹性结构蛋白和微管蛋白等。结论 四角蛤蜊60%、80%醇沉部位具有降糖作用，可作为降糖产品进一步研究开发。Abstract: OBJECTIVE To screen the effective anti-hyperglycemic fraction of Mactra veneriformis， determine and identify polysaccharide and protein components in different alcohol precipitate fraction of Mactra veneriformis. METHODS With the blood glucose concentration of mice with alloxan-induced diabetes mellitus as screening index， then the blood was taken by removalling eyeball and INS， TG， T-CHO were determined; the content of polysaccharide and protein were determined by the anthrone-sulfuric and BCA method； the proteins were checked through SDS-polyacrylamide gel electrophoresis and LC-MS/MS. RESULTS Except 20% alcohol precipitate fraction， the others reduced the content of mice blood sugar， INS and TG. 80% alcohol precipitate fraction reduced T-CHO; the content of polysaccharide and protein in 20%， 40%， 60%， 80% alcohol precipitate fraction respectively were 21.45%， 45.82%， 14.61%， 22.84% and 23.56%， 16.85%， 28.60%， 36.10%；the kinds of protein included myosin， tropomyosin， actin， calmodulin， elastin and tubulin. CONCLUSION 60% and 80% alcohol precipitate fraction shows remarkable effects on reducing mice blood sugar content， which can be further developed as a hypoglycemic product.
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