摘要
目的研究适用于巨噬细胞甘露糖受体(mannosereceptor,MR)与配体亲和力检测的方法,并用以检测魔芋葡甘露聚糖与MR亲和力的大小。方法采用腹腔冲洗法获得腹腔巨噬细胞(macrophage,Mφ),经纯化、鉴定后与异硫氰酸荧光标记的甘露糖基化的牛血清白蛋白(M-FITC-BSA)孵育20min,分别加入D-甘露糖、提取、纯化的不同浓度的魔芋葡甘露聚糖以及ConA,采用荧光显微镜和流式细胞仪检测Mφ表面甘露糖受体与配体的结合情况。结果①流式细胞仪辅以荧光显微镜能很好的对MR与配体的亲和力进行定性和定量检测;②MR与配体结合后可被甘露糖、魔芋葡甘露聚糖、ConA抑制,且魔芋葡甘露聚糖对其抑制能力最强。结论流式细胞仪辅以荧光显微镜是一种简便、快捷、准确的检测MR与配体亲和力的方法;魔芋葡甘露聚糖与MR的亲和力强于甘露糖和ConA,可能靶向作用于MR调节机体免疫功能。
Objective To establish a method of measuring affinity of mannose receptor（MR） of peritoneal macrophage and its ligands,and to measure the affinity of MR and glucomannan.Methods Peritoneal macrophages are acquired through Irrigation of peritoneal cavity.After purification and identification,macrophages are incubated with fluorescent isothiocyanate（FITC）-labeled mannosylated albumin（M-FITC-BSA,Sigma A7790） for 20 minites,the affinity of MR and its ligands are measured by flow cytometry（FCM） and fluorescene microscope.Results ①The affinity of MR and its ligands can be measured by cytometry（FCM） and fluorescene microscope;②The binding of MR and its ligands can be inhibited by the existence of D-mannose, glucomanna and ConA, especially by glucomannan. Conclusion The measurement of the affinity of MR and its ligands by flow cytometry（FCM） and fluorescene microscope is a simple,quick and accurate;The affinity of glucomannan and MR is higher than that of mannose,ConA and MR,it may be used to regulate the immunological function by targeting at MR.
出处
《中国医药指南》
2011年第30期215-217,共3页
Guide of China Medicine
