摘要
目的为非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)发病机制和防治研究提供可靠的原代肝细胞脂肪变性细胞模型。方法采用Seglen两步灌流法及其改良方法分离获取小鼠原代肝细胞,体外培养48 h后,给予不同浓度的游离脂肪酸(free fatty acid,FFA)混合物(油酸∶棕榈酸=2∶1)诱导24 h,采用油红O染色,观察肝细胞内脂滴沉积情况,全自动生化分析仪测定细胞内三酰甘油(TG)含量。结果与对照组相比,模型组肝细胞内出现红色脂滴沉积,且与加入的FFA混合物浓度呈剂量依赖性;模型组肝细胞内TG含量明显高于对照组(P<0.05),且随着加入的FFA浓度升高而增加,提示肝细胞内脂质沉积随着加入的FFA浓度升高而增加。结论成功构建了原代肝细胞脂肪变性细胞模型,为NAFLD的研究提供了可靠的细胞模型。
Objective To provide a reliable primary hepatocyte steatosis cell model for the pathogenesis,prevention and treatment of non-alcoholic fatty liver disease(NAFLD).Methods Primary mouse hepatocytes were isolated by Seglen two-step perfusion method and its improved method.After cultured in vitro for 48 hours,different concentrations of free fatty acid(FFA)mixture[oleic acid(OA)∶palmitic acid(PA)=2∶1]were induced for 24 hours.The lipid droplet deposition in hepatocytes was observed by oil red O staining and the content of triglyceride(TG)in cells was determined by automatic biochemical analyzer.Results Compared with the control group,there were red lipid droplets in hepatocytes in the model group,and the concentration of FFA mixture was dose-dependent,and the content of TG in hepatocytes in the model group was significantly higherthan that in the control group(P<0.05),and increased with the increase of the concentration of FFA,suggesting that the lipid deposition in hepatocytes increased with the increase of FFA concentration.Conclusion The primary hepatocyte steatosis cell model is successfully established,which provides a reliable cell model for the study of non-alcoholic fatty liver disease.
作者
梁蕊
滕景芳
李钊
徐文斌
孟明
LIANG Rui;TENG Jingfang;LI Zhao;XU Wenbin;MENG Ming(School of Basic Medicial Sciences,Hebei University,Baoding 071000,China;Key Laboratory of Pathogenesis Mechanism and Control of Inflammatory-Autoimmune Diseases of Hebei Province,Baoding 071000,China)
出处
《医学研究与教育》
CAS
2021年第2期1-7,共7页
Medical Research and Education
基金
国家自然科学基金面上项目(81771755)
河北省自然科学基金(H2020201022,H2020201300)。
关键词
非酒精性脂肪性肝病
原代肝细胞
肝脂肪变性细胞模型
nonalcoholic fatty liver disease(NAFLD)
primary hepatocytes
hepatic steatosis cell model
