不同诱导极化方式对大鼠骨髓来源巨噬细胞增殖、凋亡及吞噬能力的影响  

Effects of different induced polarization methods on the proliferation,apoptosis and phagocytosis of rat bone marrow-derived macrophages

作　　者：李莉 卓瑾[1,2] 郑玲 周玲 王启松 罗岚 骆凯[2] Li Li;Zhuo Jin;Zheng Ling;Zhou Ling;Wang Qisong;Luo Lan;Luo Kai(Fujian Key Laboratory of Oral Diseases&Fujian Provincial Engineering Research Center of Oral Biomaterial&Stomatological Key Laboratory of Fujian College and University,Fuzhou 350002,Fujian Province,China;Institute of Stomatology,Fujian Medical University&Center of Oral Tissue Engineering,Fujian Medical University&Hospital of Stomatology,Fujian Medical University,Fuzhou 350002,Fujian Province,China;Fujian Armed Police Corps Hospital,Fuzhou 350003,Fujian Province,China;Fujian Provincial Governmental Hospital,Fuzhou 350003,Fujian Province,China)

机构地区：[1]福建省口腔疾病研究重点实验室,福建省口腔生物材料工程技术研究中心,福建省高校口腔医学重点实验室,福建省福州市350002 [2]福建医科大学口腔医学研究院,福建医科大学口腔组织工程研究中心,福建医科大学附属口腔医院,福建省福州市350002 [3]福建武警总队医院,福建省福州市350003 [4]福建省级机关医院,福建省福州市350003

出　　处：《中国组织工程研究》2021年第25期4032-4037,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81870766),项目负责人:骆凯;福建省科技创新联合资金项目(2016Y9023),项目负责人:骆凯;福建省卫生计生委青年科研项目(2016-1-72),项目负责人:罗岚。

摘　　要：背景:经典激活巨噬细胞和替代激活巨噬细胞具有不同的表型和功能,目前尚鲜有研究探讨不同诱导方式对骨髓来源巨噬细胞生物学功能的影响。目的:探讨不同诱导极化方式对大鼠骨髓来源巨噬细胞生物学行为的影响。方法:通过原代细胞形态学观察、瑞氏染色和流式细胞术对体外分离培养的大鼠骨髓来源巨噬细胞进行鉴定。经γ-干扰素和脂多糖、白细胞介素4分别诱导激活24 h,倒置显微镜观察细胞形态,Real time-PCR检测细胞表面标记物Nos2和Arg1的表达,CCK-8法检测细胞增殖能力,流式细胞术检测细胞凋亡水平,中性红染色鉴定细胞吞噬功能。结果与结论:经γ-干扰素和脂多糖联合刺激的骨髓来源巨噬细胞高表达M1型细胞表面标记物Nos2,可促进细胞凋亡并且降低细胞的吞噬功能;而经白细胞介素4刺激的骨髓来源巨噬细胞则高表达M2型细胞表面标记物Arg1,提高细胞增殖和吞噬能力,抑制细胞凋亡。结果表明,不同诱导极化方式可影响骨髓来源巨噬细胞的生物学行为。BACKGROUND:Classically activated and alternative activated macrophages have different phenotypes and functions,and there are few studies to explore the effect of different induction methods on the biological functions of bone marrow-derived macrophages.OBJECTIVE:To explore the effect of different induced polarization ways on biological behaviors of bone marrow-derived macrophages from rats.METHODS:Rat bone marrow-derived macrophages were isolated and cultured in vitro,and identified by morphological observation,Wright Giemsa staining and flow cytometry.After being induced and activated by interferon-γ,lipopolysaccharide and interleukin-4 for 24 hours,the morphology of bone marrowderived macrophages was observed by inverted microscope.The expression levels of cell surface markers Nos2 and Arg1 were detected by real time-PCR.CCK8 assay was performed to determine the proliferation ability of bone marrow-derived macrophages.Flow cytometry was performed to detect the apoptosis,and neutral red staining was performed to identify the phagocytic function of bone marrow-derived macrophages.RESULTS AND CONCLUSION:Bone marrow-derived macrophages that stimulated by interferon-γand lipopolysaccharide were highly expressing the cell surface marker Nos2 of M1,and apoptosis of bone marrow-derived macrophages can be slightly promoted as well as the phagocytic function of bone marrow-derived macrophages is reduced;while bone marrow-derived macrophages stimulated by interleukin-4 are highly expressing the cell surface marker Arg1 of M2.Cell proliferation and phagocytosis of bone marrow-derived macrophages are promoted,and apoptosis of bone marrow-derived macrophages is inhibited.The results confirm that the biological behavior of bone marrow-derived macrophages can be affected by different methods of induced polarization.

关 键 词：骨髓 巨噬细胞 1型巨噬细胞 2型巨噬细胞 极化 Γ-干扰素 脂多糖 白细胞介素4 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]