两种细胞系用于建立体外脂肪肝模型比较.docVIP

两种细胞系用于建立体外脂肪肝模型比较 　 作者：潘雪丰，温彩霞，许建华 【摘要】 目的 对比油酸诱导肝癌细胞株HepG2、正常肝细胞株L02建立的肝细胞脂肪变性模型,通过检验复方蛋氨酸胆碱的防治作用，寻找简便、稳定的体外药物筛选模型。方法 设正常组、HepG2模型组、HepG2不同浓度给药组、L02模型组、L02不同浓度给药组、用油酸诱导肝细胞脂肪变,复方蛋氨酸胆碱干预，以油红O染色镜下观察细胞内脂滴形成状况,并检测细胞上清中的ALT、AST、γGT水平。结果 油酸刺激HepG2、L02 肝细胞24 h,细胞内典型脂肪滴形成。复方蛋氨酸胆碱2次给药，药物难以使HepG2细胞内脂滴减少，L02细胞在加油酸48 h后油红O 染色证明细胞内脂滴可减少，脂滴表达的红色IOD值，各给药组与模型组比较，差异有极显著性(Plt;0.01)。细胞上清液中ALT、AST、γGT，复方蛋氨酸胆碱各组与模型组比较差异有显著性，证实其对肝细胞损伤较小。结论 L02比HepG2更适合作为体外的筛药细胞模型，油酸刺激L02细胞形成脂肪变，药物提前24 h干预，24 h后再给药1次，这种造模和给药方案可作为一种可行的脂肪肝细胞筛药模型使用。 【关键词】 肝癌细胞株HepG2;L02肝细胞株;油酸;脂肪肝;模型;体外;复方蛋氨酸胆碱 Abstract:Objective To compare two models of hepatocytic steatosis in vitro，which were established by treating hepatocarcinoma cell HepG2 and human liver cell L02 with oleic acid in vitro. Through examination the role of compound methionine and choline bitartrate,to find a convenient and stable model in vitro for drug screening.Methods Normal group,HepG2 model group,groups of HepG2 cells treated with compound methionine and choline bitartrate at different concentrations,L02 model group,and model groups of L02 cells treated with drug at different concentratrions were set up. The hepatocytic steatosis models were established by treating HepG2 and L02 with oleic acid in vitro. The fatty droplets in cells could be observed by oil red O staining under light microscope. ALT,AST,and γGT in cell supernatant were detected by reactant kits.Results After treated with oleic acid for 24 hours,the fatty droplets in cells were observed by oil red O staining under light microscope. With the prevention with compound methionine and choline bitartrate,the amount of lipid droplet in HepG2 cells did not decrease,but decreased in L02 cells after treated with oleic acid for 48 hours. Compared with the model group,the integral optical density (IOD) value of positive of red fatty droplets in the group treated with drugs decreased markedly (Plt;0.01). There were significant differences in ALT, AST, and γGT in cell supernatant in the group