藏药蕨麻对原代培养小鼠肝细胞酒精损伤保护作用研究.PDFVIP

藏药蕨麻对原代培养小鼠肝细胞酒精损伤保护作用研究  罗慧英 1,2 ，曹茸茸 1 ，黄亚红 1(1.甘肃中医药大学药学院药理教研室，兰州 730000 ；2.甘肃省中药药理与毒理重点实验室，兰州 730000) 摘要：目的 研究藏药蕨麻对原代培养酒精损伤肝细胞的保护作用。方法 原代肝细胞经分离纯化培养后，MTT 法评价 藏药蕨麻对酒精损伤肝细胞存活率的影响；荧光染色法测定藏药蕨麻对酒精损伤肝细胞活性氧物质(ROS)含量、细胞内钙 离子浓度的影响；流式细胞仪检测藏药蕨麻对酒精损伤肝细胞凋亡的影响；免疫印迹法检测藏药蕨麻对 Bcl-2 和 Bax 表 达的影响。结果 经酒精损伤后，肝细胞存活率降低；细胞内 ROS 含量和钙离子浓度增高；凋亡抑制基因 Bcl-2 减弱、 促凋亡基因 Bax 表达增强。藏药蕨麻可明显提高细胞存活率；降低细胞内 ROS 含量和钙离子浓度；改善凋亡情况，增强 Bcl-2 表达，抑制 Bax 表达，且作用呈剂量依赖性。结论 藏药蕨麻对原代培养小鼠肝细胞酒精损伤具有一定的保护作用。 关键词：藏药蕨麻；原代培养肝细胞；酒精性肝损伤 中图分类号：R285.4 文献标志码：B 文章编号：1007-7693(2016)09-1130-04 DOI: 10.13748/ki.issn1007-7693.2016.09.009 Protective Effect of Tibetan Medicine Potentilla Anserina L. on Ethanol-induced Damage in Prime Liver Cell of Mice LUO Huiying1,2 1 1 , CAO Rongrong , HUANG Yahong (1.Department of Pharmacology, Gansu University of Chinese Medicine, Lanzhou 730000, China; 2.Key Laboratory of Pharmaco logy and Toxicology for Traditional Chinese Medicines of Gansu Province, Lanzhou 730000, China) ABSTRACT: OBJECTIVE To study the protective effct of Tibetan medicine Potentilla anserina L. on ethanol-induced damage in mice prime liver cell. METHODS After isolation, purification and culture of primary hepatocytes, the livability of cell was evaluated by MTT; fluorescence microscopy were used to evaluated the contents of reactive oxygen species (ROS) and concentration of Ca2+ ; flow cytometry was used to detect the cell apoptosis; the expression of Bcl-2 and Bax were detected by Western bloting. RESULTS After damaged by alcohol, the cell livability decreased, the contents of Ros and concentration of Ca2+ increased, the expression of Bcl-2 decreased while that of Bax increased. Potentilla anserina could increase the cell livability, decreased the intracel