摘要
目的:研究游离脂肪酸(FFA)诱导的非酒精性脂肪性肝炎(NASH)细胞模型,动态观测细胞脂肪变性、炎症、过氧化损伤及糖转运缺陷发生发展的特点。方法:以HepG2细胞为实验对象,采用不同浓度游离脂肪酸(0.25mM,0.5mM,1mM)对HepG2细胞进行不同时间(12小时,18小时,24小时)的干预刺激,并通过油红O染色,酶联免疫的方法分别对细胞内脂肪变及细胞培养上清液中白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、超氧化物歧化酶(SOD)、丙二醛(MDA)、葡萄糖转运蛋白4(GLUT4)进行测定评估。结果:FFA作用的实验各组,随着时间的延长及浓度的增加,细胞脂肪样变、IL-8、TNF-α、SOD、MDA及GLUT4均有不同程度的改变,以24小时0.5mM FFA、24小时1mM FFA最为明显,与同期空白对照组相比,其在IL-8、TNF-α、SOD、MDA、GLUT4的表达方面均有统计学意义(P<0.05),油红O染色可见典型的"印戒"状。结论:应用游离脂肪酸诱导建立体外NASH模型成模效果好,重复率高,以0.5mM FFA 24小时为佳。
Objective： To establish a cell model of nonalcoholic steatohepatitis （NASH） by free fatty acid （FFA） and to dynamically monitor the pathological features of steatosis, oxidative damage and inflammation in these cells. Methods： Liver cells HepG2 were exposed to increasing concentrations of a fresh mixture of exogenous FFA （0. 25mM, O. 5mM, lmM） in molar ratio 1：2 palmitic： oleic at different time points （12h, 18h, 24h） respectively. Oil red O staining and the IL-8, TNF-ot, SOD, MDA, GLUT4 in supernatant were evaluated. Results： Steatosis could be seen in FFA groups to different extents. IL-8, TNF- ct, SOD, MDA and GLUT4 gradually differed according to the change of stimulation time by different FFA concentrations, espe- cially in the groups with 0. 5mM and lmM FFA concentrations for 24h, of which groups there could be seen a typical steatosis. Conclusion： A cell model of NASH has been created successfully by FFA with 0. 5mM for 24h.
出处
《中西医结合肝病杂志》
CAS
2013年第4期225-227,I0002,共4页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
高等学校学科创新引智计划资助项目(No.B07007)
北京市自然科学基金资助项目(No.7102094)
北京中医药大学创新团队资助项目(No.2011-CXTD-24)
