磷酸鞘胺醇对小鼠单核巨噬细胞吞噬功能的调控及机制研究

发布时间：2018-06-15 15:17

本文选题：吞噬 + 单核巨噬细胞　；参考：《中国细胞生物学学报》2015年09期

【摘要】：采用Western blot、免疫荧光和PCR检测小鼠单核巨噬细胞系RAW264.7中S1P受体1-3(S1PR1-3)的表达,然后应用吞噬实验和免疫荧光的方法检测磷酸鞘胺醇(sphingosine 1-phosphate,S1P)对其吞噬功能的调节。分别应用药理学工具和小干扰RNA的方法研究S1P调节其吞噬活性的作用机制。结果显示,小鼠单核巨噬细胞系RAW264.7表达S1PR1-3;S1P剂量依赖地增强小鼠单核巨噬细胞系RAW264.7的吞噬功能,应用S1PR2或S1PR3的拮抗剂和si RNAs可抑制S1P增强的小鼠单核巨噬细胞系RAW264.7的吞噬活性;而应用S1PR1的拮抗剂和si S1PR1并不影响S1P增强的RAW264.7的吞噬作用;且S1P可以显著上调RAW264.7中S1PR2和S1PR3的表达,但是不改变S1PR1的表达,提示S1P通过正反馈机制增强其介导的小鼠单核巨噬细胞系RAW264.7的吞噬功能。结果表明,S1P/S1PR2/3信号通路增强小鼠单核巨噬细胞吞噬活性,为单核巨噬细胞吞噬作用的分子机制调控研究提供了新线索。
[Abstract]:Western blot, immunofluorescence and PCR were used to detect the expression of S1P receptor 1-3P1PR1-3 in mouse monocyte macrophage cell line RAW264.7. The phagocytic function of sphingosine 1-phosphateS1P was detected by phagocytosis test and immunofluorescence. Pharmacological tools and small interfering RNA were used to study the mechanism of S1P regulating phagocytic activity. The results showed that the phagocytic activity of mouse mononuclear macrophage RAW264.7 was enhanced in a dose-dependent manner by using S1PR2 or S1PR3 antagonists and siRNAs. The phagocytic activity of murine mononuclear macrophage RAW264.7 was inhibited by using S1PR2 or S1PR3 antagonists. The antagonists of S1PR1 and si S1PR1 did not affect the phagocytosis of RAW264.7 enhanced by S1P, and S1P significantly up-regulated the expression of S1PR2 and S1PR3 in RAW264.7, but did not change the expression of S1PR1. The results suggested that S1P enhanced the phagocytic function of mouse macrophage RAW264.7 mediated by S1P through positive feedback mechanism. The results showed that S1P / S1PR2 / 3 signaling pathway enhanced the phagocytic activity of murine mononuclear macrophages and provided a new clue for the study of the molecular mechanism of mononuclear macrophage phagocytosis.
【作者单位】：首都医科大学基础医学院;
【基金】：国家自然科学基金(批准号:81430013) 北京市属高等学校创新团队建设与教师职业发展计划项目(批准号:IDHT20150502)资助的课题~~
【分类号】：R392.12

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