摘要
目的研究吲哚胺加双氧酶(IDO)对巨噬细胞表型转化的影响。方法以10 ng/mL佛波酯诱导THP-1人单核细胞,建立分化的巨噬细胞模型,分别以100 U/mL IFN-γ和100 ng/mL巨噬细胞集落刺激因子(M-CSF)刺激分化的THP-1细胞,获得M1型和M2型巨噬细胞,利用实时定量PCR(qRT-PCR)、流式细胞术和Western blot法检测巨噬细胞HLA-DR、CC型趋化因子受体7(CCR7)、IL-12p35、IL-10、CXCR4和IDO的表达变化。采用细胞转染实验对分化的THP-1细胞进行IDO的过表达和针对IDO基因的siRNA干扰实验,检测其IL-12p35、CCR7、IL-10和CXCR4的表达变化。结果 IFN-γ可诱导THP-1细胞向M1型发生转化并上调IDO的表达。而在巨噬细胞内过表达IDO促进THP-1向M2型极化,沉默细胞内的IDO基因则导致THP-1细胞极化为M1型。结论 IDO表达可调控巨噬细胞极化。
Objective To investigate whether indoleamine2,3-dioxygenase（IDO） has an effect on macrophage polarization of differentiated THP-1（ dTHP-1） cells. Methods The macrophage model was established through incubating the human monocyte line（ THP-1 cells） with phorbol-12-myristate 13-acetate（ PMA）（ 10 ng /mL） for 48 hours. To generate M1 /M2-polarized macrophages,dTHP-1 cells were cultured with IFN-γ（ 100 U /mL） and M-CSF（ 100 ng /mL）,respectively. The expressions of molecular markers of macrophages（ including HLA-DR,CCR7,IL-12p35,IL-10,CXCR4） and IDO were examined by real-time quantitative PCR（ qRT-PCR）,flow cytometry,and Western blotting. To investigate the role of IDO in the dTHP-1 cell polarization,the plasmid pEGFP-N1-IDO and siRNA-IDO were respectively transfected into cells. The mRNA levels of molecular markers of macrophages were examined by qRT-PCR. Results IFN-γ could induce the differentiation of THP-1 cells into M1 phenotype and up-regulate the IDO expression. Interestingly,our results also indicated that the ectopic IDO could trigger the transformation of dTHP-1 cells to M2 phenotype. In contrast,the knockdown of IDO expression in dTHP-1 cells resulted in increased M1 markers and lower M2 markers. Conclusion The expression intensity of IDO could modulate macrophage polarization in THP-1 cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第9期901-905,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家重点基础研究发展计划(973)(2011CB935803)
国家自然科学基金(81071712,81272311)
