人肝细胞系L.pdf

J South Med Univ, 2014, 34(6): 837-842 doi 10.3969/j.issn. 1673-4254.2014.06.17 ·837 · 基础研究 人肝细胞系人肝细胞系L-L-0202 细胞单纯肝脂肪变性细胞模型的建立与应用细胞单纯肝脂肪变性细胞模型的建立与应用 殷锦锦，唐外姣，曾 璐，周本杰 南方医科大学珠江医院药学部，广东 广州 510282 摘要：目的探讨建立人肝细胞L-02 单纯肝脂肪变性细胞模型的方法与模型的应用。方法 体外用1640培养基培养L-02 细胞， 分为模型组和正常组。当细胞融合达到70%~80% ，正常组换用新鲜1640培养液，而模型组改为含有游离脂肪酸（油酸和棕榈酸） 混合物的1640培养基诱导培养24 h。以油红O染色定性观察细胞内脂滴并通过显色比较各组间总脂质差别，模型鉴定采用观察 细胞内脂滴形态，检测细胞内甘油三酯和细胞培养上清丙二醛含量，并检测细胞凋亡情况；同时利用此模型观察护肝清脂片对 脂肪变性肝细胞内脂滴的影响。结果 模型组L-02 细胞内脂滴大量形成，且甘油三酯明显升高，而培养液中肝酶释放量及丙二 醛较正常组没有明显升高，且凋亡率没有明显增加；护肝清脂片含药血清两次给药，可以使细胞内脂滴和甘油三酯明显减少。 结论 采用油酸和棕榈酸混合物（油酸∶棕榈酸=2 ∶1）可以成功诱导L-02 细胞脂肪变性，该模型适用于治疗脂肪肝药物的研究。 关键词：脂肪肝；脂肪变性；L-02 细胞；细胞模型 Establishment of a L-02 cell model of hepatic steatosis YIN Jinjin, TANG Waijiao, ZENG Lu, ZHOU Benjie Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China Abstract: Objective To establish an in vitro cell model for investigating hepatic steatosis in non-alcoholic fatty liver disease. Methods L-02 cells cultured in 1640 containing 10% fetal bovine serum were divided into control group and model group. At 70%-80% confluency, L-02 cells in the model group were exposed to a long-chain mixture of free fatty acids (FFA, oleate and palmitate ) for 24 h, and cells in control group were treated with fresh medium. Lipid droplets in the cells were observed and total lipid content was determined with Oil Red O staining. The morphology of lipid droplets, trilyceride level, malonaldehyde content and cell apoptosis rate were evaluated to verify the cell model, and the effect of Huganqingzhi tablet on the lipid droplets was observed. Results A large number of lipid droplets were found in the cell model, which showed markedly increased level of triglyceride without significant changes of malonadehyde content or cell apoptosis rate. Intervention with