【摘要】 目的:研究过表达小鼠树突状细胞表面受体Fpr2对树突状细胞成熟和趋化功能的影响。方法:通过RVG-P靶向多肽介导质粒PEGFP-N1-Fpr2转染小鼠树突状细胞,上调表面受体Fpr2的表达。ELISA检测IL-6、IL-10,IL-12 p70及TNF-α的水平;通过流式细胞术检测细胞表面分子CD40,CD80,CD83,CD86,MHCII的表达;使用Transwell小室检测细胞迁移能力;Western blot检测MAPK通路(P38,ERK1/2,JNK)的磷酸化情况。结果:上调Fpr2后,表面分子CD83的表达水平明显升高。在细胞因子水平上,转染PEGFP-N1-Fpr2组的树突状细胞的IL-6,IL-10,TNF-α分泌量明显增多。转染质粒DNA后,细胞对CCL21的趋化减弱,但与转染空载体pEGFP-N1组相比,转染pEGFP-N1-Fpr2组的趋化细胞数明显增多。转染pEGFP-N1-Fpr2组的P38,JNK及ERK1/2磷酸化水平均显著高于转染pEGFP-N1组。结论:过表达细胞表面受体Fpr2能促进树突状细胞的成熟,并可能通过上调MAPK信号通路中的P38和JNK的磷酸化来增加其趋化能力。更多还原

【Abstract】 Objective: To investigate the regulatory effect of over-expression of Formyl Peptide Receptor-2( Fpr2) on dendritc cell maturation and chemotaxis. Methods: Dendritic cell line DC2. 4 was transfected with plasmid DNA pEGFP-Fpr2 to upregulate the expression of Fpr2 with the aid of a targeted peptide RVG-P. The concentration of selected cytokines( IL-6,IL-10,TNF-α,IL-12 p70) was measured by using the ELISA KIT. The expression levels of surface marker molecules( CD40,CD80,CD83,CD86,MHCII) were measured by FACS. Migration assay was performed by using Transwell migration chambers. Western blot was performed to evaluate the phosphorylated forms of intracellular signal transducers of MAPK( P38,ERK1/2,JNK). Results: The expression levels of CD83 in DC2. 4 increased upon up-regulation of Fpr2. The cytokine levels of IL-6,IL-10 and TNF-α increased after up-regulation of Fpr2. The average number of cell migration decreased after plasmid DNA transfection. However,the migration ability of pEGFP-Fpr2 transfected cells increased compared with the pEGFPN1 transfected cells. Phosphorylation levels of P38,JNK and ERK1/2 in the pEGFP-Fpr2 transfected cells were observed with a significant increase when compared with the pEGFP-N1 transfected cells. Conclusion: Over-expression of Formyl Peptide Receptor-2 promoted maturation and chemotaxis of dendritic cells.更多还原