壳寡糖结合并激活巨噬细胞机制的研究.PDFVIP

·124 · JChinMedUniv 35 2 200 6 4 壳寡糖结合并激活巨噬细胞机制的研究 1 1, 2 侯丽娜 ,赵鲁杭 (1., 3 1003 1;2.) [] :研究壳寡糖结合并激活巨噬细胞的机制。:RT-PCR和 ELISA检测壳寡糖对巨噬细胞及经 γ干扰 (IFN- γ)预刺激巨噬细胞的白介 1β (IL-1β)基因表达水平的影响;钙离子脂化探针 Fluo-3/AM检测壳寡糖对巨噬细胞胞浆游离 2 + Ca浓度的影响;用四甲基异硫氰酸罗丹明标记甘露聚糖, 激光共聚焦显微镜和流式细胞仪观察标记糖结合及进入巨噬细胞 的情况;通过竞争性抑制实验探讨巨噬细胞摄取壳寡糖的可能途径。:壳寡糖对巨噬细胞 IL-1β 基因表达的影响呈浓度 2 + 与时间依赖性, IFN-γ预刺激巨噬细胞有增强作用;高浓度壳寡糖引起巨噬细胞胞浆游离 Ca浓度升高;壳寡糖竞争性抑制巨 噬细胞内吞甘露聚糖, 抑制强度达 44%。:壳寡糖可能是经由巨噬细胞表面的甘露糖受体介导结合并激活巨噬细胞,钙 离子相关的信号转导途径可能参与这 一过程。 [] 壳寡糖;巨噬细胞;甘露糖受体;白细胞介 1β;γ干扰 [] R34 [] A [] 025 8-4646 (2006 )02-0 124-04 Bindingandstmiulatoryeffectofoligochitosaninmacrophages 1 1, 2 HOULi-na, ZHAOLu-hang (1.DepartmentofBiochemistryandMolecularBiology, SchoolofMedicine, ZhejiangUniversity, Hangzhou3 10031, China; 2.CellularandMolecular ResearchCenter) [ Abstract] Objective:Toexplorethemechanismofthebindingandstmiulatoryeffectofoligochitosaninmacrophages(MP).Meth- ods:Theeffectofoligochitosanoninterleukin-1β (IL-1β)geneexpressioninrestingmacrophagesandmacrophagesprestmiulatedwith interferon-γ(IFN-γ)wasdetectedbyusingreversetranscriptionpolymerasechainreaction(RT-PCR)andenzyme-linkedimmunosor- bentassay(ELISA).MacrophagestreatedwitholigochitosanofdifferentconcentrationsweredyedwithFluo-3/AM toobservetheeffect 2 + ofoligochitosanontheleveloffreeCa inthecytoplasm.Mannanwaslabeledwithtetramethylrhodamineisothiocyanate.Theinterac- tionsbetweenmacrophagesandthelabeledsugarwereobservedunderconfocallasermicroscopeandanalyzedbyusingflowcytometry. Competitiveinhibitionstudywasperformedtodeterminethemechanismofoligochitosanuptakebymacrophage.sResults:Oligoch-i tosanstimulatedIL-1β geneexpressionintheconcentration-dependentandtmie-dependentmanne,randthisstmiulatoryeffectwasaug-