目的 探讨SETD4（SET-domain containing protein 4）基因敲除对小鼠腹腔巨噬细胞（peritoneal macrophages，pMφ）分化成熟、功能的影响以及对小鼠外周血、脾脏免疫细胞分化，胸腺和脾脏发育的影响。 方法 (1)利用流式细胞术检测腹腔灌洗液中pMφ表面CD31分子的表达情况；(2)用液相芯片技术检测和比较SETD4-/-和SETD4+/+小鼠pMφ在受到脂多糖（LPS）刺激后对细胞因子TNF-α、IL-6释放的影响, 流式细胞术检测SETD4基因敲除对巨噬细胞吞噬细菌能力及Transwell检测对巨噬细胞迁移能力的影响；(3)流式细胞术分析和比较两组小鼠外周血及脾脏主要免疫细胞的数量及比例的差异；(4)HE染色对比两组小鼠胸腺和脾脏组织结构的差异。 结果 （1）与SETD4+/+小鼠相比，SETD4-/-小鼠pMφ受到LPS刺激后TNF-α、IL-6的释放明显减少，但是两组小鼠pMφ的比例及分化成熟程度无明显差异；（2）吞噬细菌能力和迁移能力两组无明显差异；（3）两组小鼠外周血及脾脏主要免疫细胞的数量及比例无明显差异；（4）SETD4基因敲除对小鼠胸腺和脾脏的结构无明显影响。 结论 （1）SETD4能促进炎性细胞因子TNF-α、IL-6的产生，但对pMφ的分化成熟及细菌吞噬、迁移能力无明显影响；（2）SETD4基因敲除对小鼠外周血及脾脏免疫细胞的分化没有明显影响，对胸腺和脾脏组织的发育无明显影响。
Abstract
Objective To investigate the effects of SETD4 (SET-domain containing protein 4) knockout on the differentiation, maturity and function of pMφ (peritoneal macrophages), differentiation of immune cells in peripheral blood and spleen, and development of thymus and spleen in mice. Methods The expression of CD31 on pMφ was detected by flow cytometry. The release of both TNF- α and IL-6 from LPS-treated pMφ in SETD4-/- and SETD4+/+ mice were examined and compared by liquid chip technology. The phagocytosis of pMφ was detected by flow cytometry and the migration of pMφ was measured by Transwell system. The number and proportion of immune cells in blood and spleen from SETD4-/- and SETD4+/+ mice were detected by flow cytometry. Difference on the structure of thymus and spleen from SETD4-/- and SETD4+/+ mice was compared by HE staining. Results (1)Compared with SETD4+/+ cells, the release of TNF- α and IL-6 decreased in SETD4-/- pMφ in response to LPS stimulation, while there were no significant difference in ratio, differentiation and maturity of pMφ. (2)There were no significant difference in phagocytosis and migration between SETD4-/- and SETD4+/+ pMφ. (3)There were no significant difference in number and proportion of immune cells in peripheral blood and spleen from SETD4-/- and SETD4+/+ mice. (4)SETD4 knockout had no significant effect on the structure of thymus and spleen from SETD4-/- and SETD4+/+ mice. Conclusions (1)SETD4 has a positive role in the production of TNF- α and IL-6 from LPS stimulated pMφ, while no significant effect on the differentiation, maturity, phagocytosis and migration of pMφ. (2)SETD4 knockout has no obvious effect on the differentiation of immune cells in peripheral blood and spleen, and the development of thymus and spleen.
关键词
SETD4 /
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免疫细胞 /
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巨噬细胞 /
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细胞因子
Key words
SETD4 /
Immune cells /
Peritoneal macrophages /
Inflammatory cytokines
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基金
国家自然科学基金项目（81471901）；广州市科学计划项目（201904010382）
京ICP备08002354号-3
