小鼠骨髓和腹膜腔来源巨噬细胞的功能差异  

The functional differences of macrophages derived from murine bone marrow and peritoneal cavity

作　　者：黄镘静 罗扬淦 卢姿含 王伟玲 郝逗逗 廖晗婧 朱枝祥 HUANG Manjing;LUO Yanggan;LU Zihan;WANG Weiling;HAO Doudou;LIAO Hanjing;ZHU Zhixiang(School of Chinese Materia Medica,Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Modern Research Center of Traditional Chinese Medicine,Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院,北京100029 [2]北京中医药大学北京中医药研究院中药现代研究中心,北京100029

出　　处：《细胞与分子免疫学杂志》2024年第3期215-221,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81603361)。

摘　　要：目的研究骨髓来源巨噬细胞和腹膜腔巨噬细胞的功能差异,为免疫学研究和免疫调节药物评价中巨噬细胞的选择提供依据。方法利用巨噬细胞集落刺激因子(M-CSF)诱导骨髓单核细胞分化为巨噬细胞,利用巯基乙酸盐培养基诱导腹膜炎获得腹膜腔巨噬细胞,两种巨噬细胞在分别经过酵母多糖(zymosan)、脂多糖(LPS)、雷西莫特(R848)和非甲基化的胞嘧啶鸟嘌呤二核苷酸寡聚体(CpG)四种Toll样受体(TLR)配体诱导后,实时荧光定量PCR测定巨噬细胞肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、巨噬细胞炎性蛋白1α(MIP-1α)和单核细胞趋化蛋白1(MCP-1)mRNA水平,ELISA检测巨噬细胞培养上清液TNF-α、IL-6、MIP-1α、MCP-1含量,流式细胞术分析细胞表面共刺激分子CD80、CD86、CD40和主要组织相容性复合体Ⅱ(MHCⅡ)的表达。结果两种巨噬细胞在经过不同TLR配体诱导后,炎症因子和趋化因子的mRNA表达水平均有不同程度的增加,腹膜腔巨噬细胞TNF-α、IL-6、MIP-1α、MCP-1分泌水平以及表面CD86和MHCⅡ表达显著高于骨髓来源巨噬细胞。结论骨髓来源巨噬细胞和腹膜腔巨噬细胞均表达炎症因子、趋化因子、共刺激分子和MHCⅡ,腹膜腔巨噬细胞具有更加完整的巨噬细胞功能,在免疫学研究和免疫调节药物评价中可以优选腹膜腔巨噬细胞。Objective To compare the functional differences between bone marrow derived macrophages and peritoneal macrophages,which may provide the basis for the selection of macrophages in immunological research and immunoregulatory drug evaluation.Methods Marophage colony-stimulating factor(M-CSF)was used to induce the differentiation of bone marrow monocytes into macrophages,and thioglycolate medium was used to induce peritonitis to obtain peritoneal macrophages.After both macrophages being stimulated by zymosan,LPS,R848 and CpG respectively,mRNA levels of tumor necrosis factorα(TNF-α),interleukin 6(IL-6),macrophage inflammatory protein 1α(MIP-1α),monocyte chemoattractant protein 1(MCP-1)were measured by Real-time fluorescent quantitative PCR and the concentrations of secreted TNF-α,IL-6,MIP-1αand MCP-1 were detected by ELISA.In addition,the expression of costimulatory molecules CD80,CD86,CD40 and histocompatibility complexⅡ(MHCⅡ)on the cell surface was analyzed by flow cytometry.Results After inducing by different TLR ligands,mRNA expression levels of inflammatory cytokines and chemokines were increased in both macrophages.The secretion of TNF-α,IL-6,MIP-1αand MCP-1 in peritoneal macrophages and the expression of CD86 and MHCⅡon the surface of peritoneal macrophages were significantly higher than those of bone marrow derived macrophages.Conclusion There are significant differences in the expression of inflammatory factors,chemokines,costimulatory molecules,and histocompatibility complex between bone marrow derived macrophages and peritoneal macrophages.Peritoneal macrophages have more complete macrophage function and is more suitable for immunological research and immunomodulatory drug evaluation.

关 键 词：单核细胞 巨噬细胞 功能差异 Toll样受体(TLR)配体 

分 类 号：R364.5[医药卫生—病理学] R392.12[医药卫生—基础医学]