血管活性肠肽对RAW264.7小鼠巨噬细胞TREM-1功能的影响

doi:10.13418/j.issn.1001-165x.2019.01.012

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (1): 55-59.doi: 10.13418/j.issn.1001-165x.2019.01.012

血管活性肠肽对RAW264.7小鼠巨噬细胞TREM-1功能的影响

宋卓慧，　李淑芬，　刘燕，　原丽，　韩玲娜，　张翠英

长治医学院基础医学部生理学系，山西长治 046000

收稿日期:2018-09-11 出版日期:2019-01-25 发布日期:2019-02-20
通讯作者: 张翠英，教授，E-mail: zhangcy157@sohu.com
作者简介:宋卓慧(1981-)，女，山西长治人，硕士，讲师，主要研究方向：急性肺损伤，E-mail: szh.11.com@163.com
基金资助:
山西省卫生计生科研课题（2015159）；长治医学院科技启动基金项目（QDZ201632）

Effects of vasoactive intestinal peptide on the function of triggering receptor expressed on myeloid cells-1 in RAW264.7 murine macrophages

SONG Zhuo-hui, LI Shu-fen, LIU Yan, YUAN Li, HAN Ling-na, ZHANG Cui-ying

Department of Physiology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China

Received:2018-09-11 Online:2019-01-25 Published:2019-02-20

摘要/Abstract

摘要：

目的　观察血管活性肠肽(VIP)对RAW264.7小鼠巨噬细胞髓样细胞表达的触发受体-1（TREM-1）功能的影响。方法　以小鼠巨噬细胞系RAW264.7为研究对象，细胞接种于培养板后采用TREM-1的单克隆激动抗体激活TREM-1，VIP预处理组采用10 nM的VIP进行干预。1 h后采用Western blot法检测胞内PLC-γ磷酸化改变，采用DCFH-DA荧光探针检测胞内活性氧簇（ROS）的含量；6 h后，采用real-time PCR法检测巨噬细胞TREM-1受体激活后下游靶基因肿瘤坏死因子-α（TNF-α）和单核细胞趋化蛋白-1（MCP-1）的基因表达；12 h后，采用ELISA法检测细胞培养上清液TNF-α和MCP-1的蛋白含量。结果　采用TREM-1单克隆激动抗体处理小鼠巨噬细胞后，可显著增高胞内PLC-γ磷酸化水平、ROS含量，以及TNF-α和MCP-1的mRNA与蛋白水平，而VIP预处理可明显抑制单克隆激动抗体所诱导的上述反应。结论　VIP可抑制RAW264.7小鼠巨噬细胞TREM-1激活后诱导的功能改变，从而发挥抗氧化抗炎的作用。

关键词: 血管活性肠肽, 　髓样细胞表达的触发受体-1, 　巨噬细胞, 　炎症

Abstract:

Objective　To observe the effect of vasoactive intestinal peptide (VIP) on the function of triggering receptor expressed on myeloid cells-1 (TREM-1) in mouse macrophages.　Methods　The mouse macrophage cell line RAW264.7 was used as the research object. After cells were seeded on the culture plates, TREM-1 was activated by the monoclonal agonistic antibody of TREM-1, and cells in the VIP pretreatment group was treated with 10 nM VIP. After 1 h, the intracellular PLC-γ phosphorylation and intracellular reactive oxygen species (ROS) were detected by Western blot and DCFH-DA, respectively. After 6 h, real-time PCR was used to detect the gene expression of tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) as the downstream target genes of TREM-1 activation. After 12 h, TNF-α and MCP-1 contentration in the cell culture supernatants were detected by ELISA. Results After treatment of TREM-1 monoclonal agonistic antibody, the intracellular PLC-γ phosphorylation level, ROS content, and mRNA and protein levels of TNF-α and MCP-1 were significantly increased in murine macrophages, while VIP pretreatment significantly inhibited the above reactions induced by monoclonal agonistic antibodies. Conclusion VIP can inhibit the functional changes induced by the activation of TREM-1 in murine macrophages RAW264.7, thereby exerting anti-oxidant and anti-inflammatory effects.

Key words: Vasoactive intestinal peptide, 　Triggering receptor expressed on myeloid cells-1, Macrophages, 　Inflammation

宋卓慧,李淑芬,刘燕,原丽,韩玲娜,张翠英. 血管活性肠肽对RAW264.7小鼠巨噬细胞TREM-1功能的影响[J]. 中国临床解剖学杂志, 2019, 37(1): 55-59.

SONG Zhuo-hui, LI Shu-fen, LIU Yan, YUAN Li, HAN Ling-na, ZHANG Cui-ying. Effects of vasoactive intestinal peptide on the function of triggering receptor expressed on myeloid cells-1 in RAW264.7 murine macrophages[J]. Chinese Journal Of Clinical Anatomy, 2019, 37(1): 55-59.

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血管活性肠肽对RAW264.7小鼠巨噬细胞TREM-1功能的影响

Effects of vasoactive intestinal peptide on the function of triggering receptor expressed on myeloid cells-1 in RAW264.7 murine macrophages

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