胶体金-荧光素双标配体法研究细胞低密度脂蛋白受体的功能

 目的建立一种利用胶体金-荧光素双标记配体研究细胞低密度脂蛋白受体(LDLR)功能的方法。方法①先用荧光素DiI标记LDL,再在一定pH值下将DiI-LDL吸附在胶体金颗粒表面上,获得稳定的双标记配体Au-DiI-LDL。②利用双标配体Au-DiI-LDL结合荧光显微镜、流式细胞仪和透射电镜等技术分别对经LDLR表达诱导剂、抑制剂或姜黄素作用后的ANA-1细胞LDLR进行定性定量分析以及内吞作用的直观检测。结果荧光显微镜、流式细胞仪和透射电镜的检测结果均表明,能够利用双标配体Au-DiI-LDL检测ANA-1细胞LDLR的活性(P<0.05)。结论胶体金-荧光素双标记配体法是一种可用于细胞LDLR功能检测的新型高效的方法。

Study on LDL Receptor Function Using Colloid Gold-Fluorescein Double-Labeled Ligand Method

OBJECTIVE To establish a method for investigating the function of LDL receptor using the double labeled of ligand tagged with the colloid gold and the fluorescein in cell.METHODS ①LDL was labeled by a fluorescent reagent,DiI.Then the DiI-LDL was adsorbed on the colloid gold surface under some pH value and the double labeled of ligand Au-DiI-LDL was obtained.②Using the double labeled ligand,the effects of LDL receptor gene expression inductor inhibitor or curcumin on the function of LDL receptor in ANA-1 cells were measured by fluo-microscopy(FM),fluorescence flow cytometric methods(FFCM)and transmission electron microscope(TEM).RESULTS The function of LDL receptor in ANA-1 cells were measured successfully by using the Au-DiI-LDL(P<0.05).CONCLUSION The colloid gold and fluorescein double-labeled ligand method is a new,high-performance ways to study the function of LDL receptor in cells.