体外肝摄取高密度脂蛋白(HDL)细胞模型的建立

Establishment of cellular high density lipoprotein( HDL) uptake model in vitro

目的:建立肝摄取HDL的体外细胞模型,用于促进胆固醇逆转运活性成分的高通量筛选。方法:以Di I(1,1-Dioctadecyl-3,3,3',3'-tetramethylindocao cyanine per chlorate)荧光染料标记HDL,体外培养人HepG2肝细胞株,细胞贴壁后与2μg/ml DiIHDL于37℃下恒温孵育,在动态细胞观察系统下连续观察细胞摄取HDL的实时状态,摄取完成后利用荧光显微镜照相并采用流式细胞术测定细胞的荧光值。结果:荧光显微镜和流式细胞术结果显示,5 h后HepG2细胞摄取HDL饱和。结论:通过体外培养HepG2细胞摄取Di I标记HDL建立的细胞模型并利用流式细胞术测定荧光值,可用于促进胆固醇逆转运活性成分的高通量筛选。

Objective： To promote high throughput screening of the active ingredient in reversing cholesterol transport,the hepatocyte uptake of HDL in vitro was established. Methods： The high density lipoprotein was labeled by DiI fluorescent dyes and human liver cell line HepG2 was cultured in vivo. The cells were incubated with 2 μg/ml Di I-HDL at 37℃ for continuous observation by a dynamic cell observation system. The cellular fluorescence values were measured by fluorescence microscopy and flow cytometry. Results： Fluorescence microscopy and flow cytometry showed that HepG2 cells received saturated HDL after 5 hours. Conclusion： The establishment of cell models and flow cytometry could be used in high throughput screening for reversing cholesterol transport.