甲嘎松汤对肝损伤的保护作用研究  

Study on protective effect of Jiagasong Tang for liver injury animal model and cell oxidative stress model in vitro

作　　者：熊天琴[1] 李红侠[1] 陈元元[1] 赵玉民[1] 

机构地区：[1]广州中医药大学.国家中医药管理局中药药理三级科研实验室,广州510006

出　　处：《中药药理与临床》2013年第1期132-135,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(81202997);广东省科技计划项目(2011B031700075);广东省自然科学基金(S2011010003925)

摘　　要：目的:研究藏药甲嘎松汤对整体动物急慢性肝损伤模型及体外L02细胞氧化应激损伤模型的保护作用,并进行初步机制研究。方法:制备5种小鼠急性肝损伤模型、1种大鼠慢性肝损伤模型,观察甲嘎松汤对动物血清AST、ALT活性及四氯化碳致小鼠急性肝损伤的肝组织中CAT、GSH活性;采用RT-PCR技术,测定慢性酒精性肝损伤肝脏缺氧诱导因子-1 mRNA的表达。先给予Nrf2激活剂t-BHQ干预12h,再给予甲嘎松汤的含药血清作用12h后,利用终浓度为100mU/ml的葡萄糖氧化酶(Glucose Oxi-dase,GO)诱导人肝细胞L02氧化应激损伤模型,检测细胞培养上清液中AST、ALT活性;四甲基氮唑(MTT)比色法检测细胞活力。结果:在6种肝损伤动物中均可见甲嘎松汤在0.66g/kg剂量下的AST、ALT活性比模型组显著降低;显著升高四氯化碳引起的小鼠急性肝损伤模型中抗氧化酶CAT、抗氧化剂GSH活性;在一定程度上降低模型组HIF-1α因子mRNA表达,但与模型组比较无统计学差异。在细胞模型中,20%甲嘎松汤含药血清组与t-BHQ组均能于不同时间点显著升高细胞活力;显著降低AST,ALT活性;与t-BHQ组比较,20%甲嘎松汤含药血清+t-BHQ组于不同时间点上述指标有明显改善,并且对单用t-BHQ作用8小时后表现出的肝细胞活力降低有改善作用。结论:甲嘎松汤有一定的保肝作用,与Nrf2激活剂t-BHQ有协同效应,能纠正Nrf2持续激活导致的细胞损伤,说明甲嘎松汤保肝作用可能与维持Nrf2在细胞内的动态平衡有关。To study the protective effect of Traditional Tibetan Medicine-Jiagasong Tang for animal acute and chronic liver injury model and L02 c611 oxidative stress model in vitro, and to preliminarily explore its mechanism. Methods： Five acut~ liver damage models and one mice SD rats chronic liver damage model were prepared ,and given different doses of Jiagasong Tang. The contents of ALT,AST in serum of all animals and the activity of CAT, GSH in the liver of the acute liver injury animals inducted by carbon tetrachloride. The mRNA expressions of hypoxia-inducible factor-1 were detected by RT-PCR technology in liver of chronic liver injury animals inducted by alcohol. Human L02 hepatocytes were intervened primarily by t-BHQ of Nrt2 activating agent, for 12 hours. Then fed with Jiagasong Tang drug-containing serum. After 12 hours, the oxidative stress hepatocyte injury model of human L02 hepatocytes was established by being cultured with 100mU/ml glueose oxidase （GO）. The AST, ALT of cell culture supematant were tested by speetrophotometer;the vitality of L02 cells was measured by MTT assay. Results： In all six kinds of liver damaged animals models, compared with animals model group, Jiagasong Tang could significantly reduced the active of AST, ALT（ P 〈 0.01, P 〈 0.05 ）at the dosage of 0.66g/kg; the activities of CAT and GSH in the carbon tetrachloride-injured liver were significantly increased（ P 〈 0; 01 ） ;the expression of hypoxia inducible factor-1 were partly decreased without statistical differenees. Compared with the cell model, 20% Jiagasong Tang drug-containing serum and t-BHQ groups at different time points all can significantly reduce the AST, ALT activity （ P 〈 0.01 ）, significantly increased the cell vitality （ P 〈 0.01 ） ; compared to t-BHQ group, ＂20% Jiagasong Tang drug-containing serum ＋ t-BHQ＂ group at different time points all can significantly reduce the AST, ALT activity（P 〈 0.01 ）, significantly increased the cell vitality（ P 〈 0.01 ）, and can. imp

关 键 词：甲嘎松汤 t-BHQ 人肝细胞 氧化应激 

分 类 号：R29[医药卫生—民族医学]