乙醇对小鼠原代培养肝细胞损伤及黄芩茎叶总黄酮保护作用的实验研究.docVIP

乙醇对小鼠原代培养肝细胞损伤及黄芩茎叶总黄酮保护作用的实验研究 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：乙醇对小鼠原代培养肝细胞损伤及黄芩茎叶总黄酮保护作用的实验研究 1 1 材料与方法 2 2 结果 4 3 讨论 5 文2：秦皮提取物对小鼠急性肝损伤保护作用的实验研究 6 1 材料与方法 7 2 结果 8 3 讨论 8 参考文摘引言： 9 原创性声明（模板） 11 文章致谢（模板） 11 正文 乙醇对小鼠原代培养肝细胞损伤及黄芩茎叶总黄酮保护作用的实验研究 文1：乙醇对小鼠原代培养肝细胞损伤及黄芩茎叶总黄酮保护作用的实验研究 The Protective Effect of Total Flavonoid in scutellaria baicaleis Stem-Leaf on Primary Cultured Mice Hepatocytes Injured by Ethanol Abstract：ObjectiveTo study the protective effect of total flavonoid in Scutellaria baicaleis stem-leaf (SSTF) on primary mice hepatocyes injured by ethanol. MethodsPrimary hepatocytes were isolated, cultured by trypsin digestion and induced by ethanol. Various concentratio of SSTF were added to the primary cultured hepatocytes for content of malondialdehyde(MDA) in hepatocytes and the activity of ALT in cultural supernatant were determined by general methods,the viabilities of hepatocytes were measured by elevation of MDA content of hepatocytes and ALT level in supermatant of cultural hepatocyes were restored remarkably by SSTF, hepatocytes viability was improved significantly by has a protective action on primary mice hepatocyes injured by might be associated with its anti-lopoperoridation. Key words：Ethanol; Hepatocytes; Primary cultured; SSTF; Protective effect 肝脏是酒精代谢的主要脏器，过量饮酒可造成肝脏不同程度的损害，并进一步 发展 为脂肪肝、肝纤维化和肝硬化，严重危害人类健康。了解酒精性肝损伤的发病机制，筛选有效预防和 治疗 的药物应用于临床，是当前面临的重要课题。黄芩茎叶总黄酮(SSTF)是我院中药所对黄芩的茎叶部分提取的有效成分，整体实验已证实其对肝损伤有保护作用［1］。本文建立体外乙醇性肝细胞损伤模型，在细胞水平上进一步研究乙醇性肝损伤的机制及SSTF对肝细胞保护作用机制。 1 材料与方法 药品与试剂SSTF（含总黄酮73%）：承德医学院中药研究所提供，临用前以蒸馏水配制，用饱和碳酸氢钠调pH=；Hanks液高压灭菌,4℃保存； g#12539;L-1胰蛋白酶：用Hanks液溶解，过滤除菌，调pH ，分装,-30℃保存；基础培养液：RPMI 1 640培养基 g,用超净水900 ml溶解,%NaHCO3调pH至，定溶到1 000 ml,过滤除菌，临用前每80 ml,加入新生小牛血清20 ml,青霉素100 u#12539;ml-1,链霉素100 μg#12539;ml-1,胰岛素5 μg#12539;ml-1；MTT：SIGMA公司；ALT，MDA测定试剂盒:南京建成生物工程研究所。 动物昆明种小鼠，1~3 d龄乳鼠，雌雄不限，清洁级，承德医学院实验动物管理中心提供。 仪器CO2培养箱（Taibai LNA-122D 日本），MD-100半自动生化分析仪（美国Bayer公司），721W微机型分光光度计（上海光学仪器五厂），离心机（ 上海） 肝细胞分离培养［2