琐琐葡萄多糖对大鼠体外免疫性肝损伤保护作用探究.doc

琐琐葡萄多糖对大鼠体外免疫性肝损伤保护作用探究 　 作者：刘涛 马龙 赵军 苏德奇 【摘要】 目的研究琐琐葡萄多糖（VVP）在体外对大鼠免疫性肝损伤的保护作用。方法采用热水提取、乙醇沉淀，Sevage法除蛋白、透析分离纯化提取VVP，苯酚硫酸法测定多糖含量。采用BCG整体致敏，LPS离体攻击法建立大鼠原代肝细胞免疫损伤模型，取细胞上清液测定其ALT，AST活性和NO含量。结果琐琐葡萄中多糖含量为8.19％；VVP 5，20，80μg/ml均可程度不同地使BCG+LPS损伤后升高的大鼠肝细胞上清ALT，AST活性和NO含量明显下降，其作用与阳性对照药物甘立欣（Grz）相似。结论VVP在体外对大鼠免疫性肝损伤具有明显的保护作用。 【关键词】 琐琐葡萄； 多糖； 肝细胞； 免疫性肝损伤 Abstract：ObjectiveTo study the effect of polysaccharide isolated form Vitis viniferal on immunological liver injury model in vitro.MethodsVitis amurensis Polysaccharide was obtained by extracting with water, precipitation with 95% ethanol, clearing protein with the sevage assay, and dialyzing and phenol - sulphate colorimetry was used to determine its content. The immunological hepatotoxicity model was induced by Bacillus Calmette Guerin (BCG) plus lipopolysaccharides (LPS) in vitro. The supernatant of cells after 3～24 h incubation with VVP were taken for the estimation of ALT, AST and NO respectively. ResultsThe content of polysaccharide in Vitis amurensis was 8.19％. Elevated ALT, AST activities and NO level were decreased after treatment with VVP (5，20，80 μg/ml) in different doses. The effects of VVP mentioned above were similar to those of the standard drug, Dinmmonium Glycyrrhizinate (Grz) . ConclusionVitis viniferal polysaccharide shows significant protective effects against immunological liver damage induced by BCG plus LPS in vitro. Key words：vitis viniferal; Polysaccharide; Hepatocyte; Immunological liver injury 琐琐葡萄为葡萄科葡萄属植物琐琐葡萄Vitis viniferal L.的成熟果实，主产于新疆吐鲁番、和田、鄯善等地，是维吾尔医习用的一种药食兼用植物，药用果实，具有健脾胃、理肺、生津、养血等作用。维医用于治疗脾胃不和、神志不安等，民间用于小儿麻疹、肝炎等症［1］。琐琐葡萄中除含有大量的葡萄糖、果糖、维生素、氨基酸等营养成分外，还有鞣质类、黄酮、萜类、多糖等多种生物活性物质［2～4］。研究表明多糖（polysaccharides）具有增强免疫、抗氧化、抗肿瘤、降血脂、抗衰老、抗辐射等多方面的药理作用［5］，是近年来天然药物研究的热点课题之一。为探讨多糖与琐琐葡萄抗病毒免疫调节活性的相互关系，本研究采用热水提取、乙醇沉淀，Sevage法除蛋白、透析分离纯化提取了琐琐葡萄多糖（VVP），苯酚硫酸法测定多糖含量，并采用BCG整体致敏，LPS离体攻击法建立大鼠原代肝细胞免疫损伤模型，探讨了VVP在体外对大鼠免疫性肝损伤的保护作用。 1 器材 1.1 动物SD大鼠，雄性，体重（200±20）g，新疆医科大学实验动物中心提供（SPF级，动物使用许可证号：SYXK［新］2003-0001）。 1.2 药材 琐琐葡萄购自新