降血脂体外评价技术的建立  

Establishment of a model for evaluating hypolipidemic effect in HepG2 cells

作　　者：牛玉存[1] 吕娜[1] 李颖[1] 赵丹[1] 孙长颢[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院营养与食品卫生学教研室,哈尔滨150086

出　　处：《卫生研究》2010年第2期155-158,共4页Journal of Hygiene Research

基　　金：国家"十一五"科技支撑计划项目(No.2006BAD27B01;No.2006BAD27B05)

摘　　要：目的建立一种降血脂的体外评价的方法。方法以HepG2细胞为模型,在培养液中加入胆固醇,测定细胞内胆固醇含量和胆固醇代谢相关蛋白及基因的表达情况来评价胆固醇模型是否成功,并用阳性降胆固醇药物洛伐他汀进行验证。培养液中加入游离脂肪酸,测定细胞内甘油三酯和培养液游离脂肪酸吸收率来评价模型是否成功,并用阳性降甘油三酯药物非诺贝特进行验证。结果培养液中加入25-羟胆固醇后,细胞内胆固醇含量显著升高,HMG-CoA还原酶、SREBP-2及LDLR表达降低,表明胆固醇模型成功。阳性药洛伐他汀能够显著降低细胞内胆固醇含量,上调SREBP-2及LDLR的表达,发挥降胆固醇的功效。培养液中加入50μmol油酸6h后油酸吸收率达到40%,细胞内甘油三酯有升高趋势即表明高甘油三酯模型成功。非诺贝特能够促进游离脂肪酸的吸收并且不增加细胞内甘油三酯的含量达到降低甘油三酯的作用。结论在体外可以将HepG2细胞作为细胞模型进行降血脂功能因子的筛选。Objective To establish a model of evaluating hypolipidemic effect in vitro. Methods Adding cholesterol to the culture medium for HepG2 cells to induce a hypercholesterolemia model. The content of cellular cholesterol and the expression of protein regulating cholesterol metabolism in HepG2 cells were determined. The validation of the model was identified by lovastatin,a widely used cholesterol-lowering drug. Free fatty acid was added to the culture medium for HepG2 cells to induce a hypertriglyceridemia model. The content of cellular triglyceride and the absorption rate of free fatty acid were determined. The validation of the model was identified by fenofibrate,a triglyceride-lowering drug. Results Cellular cholesterol content was increased and the expression of HMG-CoA redutase,SREBP-2 and LDLR were decreased after adding cholesterol and 25-hydrocholesterol to the culture medium. Cellular cholesterol was decreased and the expression of SREBP-2 and LDLR were up-regulated by Lovastatin. The absorption of oleic acid in cells was up to 40% after adding oleic acid （50μmol） to the culture medium for 6h. The absorption of free fatty acid was increased but the content of cellular triglyceride was not increased in cells by Fenofibrate. Conclusion This model might be an effective method for screening and assessing functional factors for lowing plasma lipids.

关 键 词：降血脂 体外评价 HEPG2 细胞 高脂血症 

分 类 号：R589.2[医药卫生—内分泌]