建立L-02肝细胞脂肪变模型方法比较及脂肪乳诱导脂变条件的优化  

Assessment on different methods for establishing hepatocyte L-02 steatosis models and optimization of the steatosis induced by medical fat emulsion

作　　者：胡启蒙[1] 陈朝银[2] 庄馨英 张旭[1] 梁杏[1] 赵声兰[1] 

机构地区：[1]云南中医学院中药学院,云南昆明650500 [2]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《中国医学物理学杂志》2015年第4期469-473,共5页Chinese Journal of Medical Physics

基　　金：国家自然科学基金(21466037);科技部科技支撑计划项目(2011BAD46B03)

摘　　要：目的:比较二甲基亚砜(DMSO)溶解油酸、乙醇溶解油酸、无水乙醇、50%胎牛血清和医用脂肪乳对L-02肝细胞的影响,优化L-02肝细胞脂肪变性模型的良好条件与方法。方法:用含10%胎牛血清PRMI-1640培养液培养细胞,细胞均匀分布进入对数生长期后随机分为正常对照组、DMSO组、DMSO+油酸组、乙醇+油酸组、乙醇组、50%胎牛血清组和医用脂肪乳组,分别用DMSO+油酸、乙醇+油酸、0.4%乙醇、50%胎牛血清(FBS)和20%医用脂肪乳培养48 h,用MTT法检测细胞活力,油红O染色观察细胞数、细胞内脂滴,用试剂盒检测细胞内甘油三酯及蛋白质。用医用脂肪乳培养L-02肝细胞,研究医用脂肪乳诱导L-02肝细胞脂肪变性的浓度和时间。结果:DMSO组和乙醇组细胞生长呈下降趋势,DMSO+油酸组和乙醇+油酸组细胞生长均呈现先升高后降低趋势。甘油三酯和蛋白质测定表明,医用脂肪乳能很好地诱导L-02肝细胞脂肪变性,医用脂肪乳含量10%,造模培养49 h,细胞脂变效果最好。结论:医用脂肪乳诱导L-02肝细胞,脂变率高、脂变细胞形态好,适合L-02肝细胞脂变模型的建立。Objective To optimize the hepatoeyte L-02 steatosis models by respectively comparing the effects of oleie acid dissolved by Dimethyl sulfoxide （DMSO）, oleie acid dissolved by ethanol, absolute ethanol, 50% fetal bovine serum （FBS） and medical fat emulsion on hepatocyte L-02. Methods Hepatocyte L-02 were cultured in PRMI-1640 supplemented with 10% FBS. When the hepatocyte L-02 distributed evenly and entered the logarithmic phase, they were randomly divided into seven groups, the normal control group and other six groups respectively cultured in oleic acid dissolved by DMSO, oleic acid dissolved by ethanol, 0.4% ethanol, 50% FBS and 20% medical fat emulsion. The hepatocyte in every group were cultured for 48 h. The viability of cells was measured by methyl thiazolyl tetrazolium （MTT） method, and the cell counts and intracellular lipid droplets could be observed by oil red O staining under optical microscope. The content of triglyceride （TG） in cells and the concentration of BCA protein were detected by corresponding detection kits. The concentration and time of hepatocyte L- 02 steatosis induced by medical fat emulsion were researched by culturing hepatocyte L-02 in medical fat emulsion. Results The cell growths in DMSO group and ethanol group showed a decreased trend, while the cell growths in oleic acid ＋ DMSO group and oleic acid ＋ ethanol group showed a firstly increased and then decreased trend. The measured results of TG and BCA indicated that the hepatocytes L-02 steatosis could be successfully induced by medical fat emulsion, and the best steatosis effect was based on 10% medical fat emulsion and the culture time of 49 h. Conclusion Medical fat emulsion is suitable to establish the hepatocytes L-02 steatosis models for it can induce hepatocytes L-02 with higher steatosis ratio and better growth morphology of steatosis cells.

关 键 词：L-02肝细胞 脂肪变性 医用脂肪乳 油酸 

分 类 号：Q28[生物学—细胞生物学] R-332[医药卫生]