人肝细胞系L-02细胞单纯肝脂肪变性细胞模型的建立与应用.pdfVIP

J South Med Un川 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L-02细胞单纯肝脂肪变性细胞模型的建立与应用 殷锦锦，房外妓，曾璐，用本杰 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筷型的应用。 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70γ俨80%，正常组换用新鲜1 640培养液，而模型组改为含有游离脂肪酸(汹酸和棕榈酸) 混合物的1640培养基诱导培养24 h。 以油红O染包定性观察细胞内B自商并通过显包比较各组间总指质差别，模型鉴定采用观察 细胞内脂滴形态，检测细胞内甘油三酶和细胞培养上消丙二瞪含量， 并检测细胞凋亡情况i 同时利用此筷型观察护肝清脂片对 脂肪变性肝细胞内脂滴的影响。 结果 筷型组L 02 细胞内脂滴大量形成，且甘油三百首明显升高， 而培养液中肝酶释放量及两二 H较正常组没有明显升高， 且凋亡率没有明显增加;护肝清脂片含药血消两次给药，可以使细胞内脂滴和甘油三百旨明显减少。 结论 采用汹酸和1棕榈酸混合物(油酸·棕榈酸 2: 1 )可以成功诱导L 02 细月刨旨肪变性，该模型适用于治疗脂肪府药物的研究。 关键词:脂肪肝;脂肪变险; 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 Be叫le Department of Phamlacy, Zht可iang Hospital, Southem Medical Universi哆 Guangzhou 510282, China Abstrad: Objective To establish al1 in vitro cell model for inves挝gating hepatic steatosis in nOI1- alcoholic fatty liver disease. Methods L-02 cells cultured in 1640 cOl1taining 10% fetal bovine 目1t11l1 were divided into control group and model group. At 70%- 80% ∞nfluency， L-02 cells in the model gr∞pwe但回归sed to a long- chain m以tu目。f free fatty acids (FFA, oleate and palmitate ) for 24 h , al1d cells in contlol gr∞p were treated w油fresh medium. Lipid droplets in the cells w町e observed and totallipid cOl1tel1t was determined with Oil Red 0 staining. The morphology of lipid dloplets, trilyceride level, malonaldehyde COl1t自lt and cell apoptosis rate were evaluated to ve1i句， the cell model, and the effect of Huganqingzhi tablet on the lipid dloplets was observed. Results A large numb臼. of lipid droplets were 旬出ld in the cell model, which showed markedly il1creased level of triglycelide without signific缸飞t changes of malol1adehyde cont.四t 0 1 cell apoptosis rate. Interv白白。11 with two doses of Huganqingzhi tablet