MTT法观察微囊藻毒素-LR对原代大鼠肝细胞的双向毒性作用  

STUDY OF THE DUALISTIC TOXICITY OF MICROCYSTIN-LR ON RAT PRIMARY HEPATOCYTES BY MTT ASSAY

作　　者：帅怡[1] 王彦琴[1] 尹建勋[2] 王俐[3] 肖萍[1] 仲伟鉴[1,2] 

机构地区：[1]上海市疾病预防控制中心,上海200336 [2]复旦大学公共卫生学院营养与食品卫生教研室 [3]上海交通大学医学院营养系

出　　处：《现代预防医学》2010年第12期2289-2291,2300,共4页Modern Preventive Medicine

基　　金：国家自然科学基金(30800935);上海市公共卫生优秀青年人才培养计划(08GWQ004)

摘　　要：[目的]系统性探讨微囊藻毒素-LR(MC-LR)引起肝细胞毒性效应的量效和时效关系特点。[方法]采用胶原酶灌注法建立原代大鼠肝细胞模型,并通过抗大鼠细胞角蛋白(CK18)的特异性抗体来进行免疫荧光染色观察细胞纯度。对分离培养得到的原代肝细胞分别给予10-5～10-12mol/l的MC-LR,在给药后的24、48、72和96h后分别采用MTT法观察细胞存货率的改变情况。[结果]分离培养所得细胞均为肝实质细胞,细胞纯度为100%;高剂量MC-LR(μM水平)能引起细胞死亡或凋亡,而低剂量MC-LR(nM^pM水平)则具有促进细胞增殖的效应,双效性作用的分界浓度10-7mol/L左右。[结论]MC-LR对原代培养肝细胞的毒性效应上表现为双向性。[Objective] To discuss the dose-response and time-response relationship of hepato-toxicity of microcystinLR（MC-LR） on rat primary hepatocytes.[Methods] Rat Primary hepatocytes were isolated by collagenase perfusion method.The purity of cells was identified by immunofluorescence using specific anti-cytokeratin 18（CK18） antibody.The isolated hepatocytes were treated with 10^-5-10^-12 mol/L of MC-LR for 24 h,48 h,72 h and 96 h,respectively,and cell survival rate was measured by MTT assay.[Results] The isolated cells were 100% parenchymal hepatocytes.High dose of MC-LR（μM） could induce cell death or apoptosis,however,low dose of MC-LR（nM-pM） enhanced cell proliferation.The threshold of MC-LR was about 10^-7 mol/L.[Conclusion] MC-LR showed the dualistic toxicity on primary hepatocytes.

关 键 词：微囊藻毒素 原代肝细胞 双效性 

分 类 号：R575[医药卫生—消化系统]