收稿日期: 2018-06-26
网络出版日期: 2018-12-29
基金资助
湖南省自然科学基金(2018JJ3368)；中南大学博士后基金（160320001）；2017年度湖南省大学生研究性学习和创新性实验计划（项目序号1003）
Triggering receptor expressed on myeloid cells-1 inhibits autophagy of macrophages under LPS stress
Received date: 2018-06-26
Online published: 2018-12-29
目的 观察髓样细胞表达的触发受体1（TREM-1）对脂多糖（LPS）应激下巨噬细胞自噬相关基因表达的影响。 方法 观察LPS应激下的巨噬细胞TREM-1蛋白的表达。分别选用TREM-1的激动剂（MAB1187）和TREM-1的拮抗剂（LR12）作用于巨噬细胞，利用qPCR检测巨噬细胞自噬相关基因ATG7、ATG5、ATG12及自噬标志蛋白微管相关蛋白1轻链3（LC3）mRNA的表达；采用Western Blot检测巨噬细胞TREM-1、LC3Ⅱ/Ⅰ、ATG7蛋白的表达；采用免疫荧光检测在LPS应激与TREM-1激活的情况下，巨噬细胞的自噬标志蛋白LC3表达。 结果 LPS（400 ng/mL、1000 ng/mL）应激下巨噬细胞TREM-1蛋白表达明显增加；LPS（1000 ng/mL）作用巨噬细胞24 h，巨噬细胞TREM-1蛋白表达达到峰值。LPS应激的巨噬细胞自噬基因ATG7、ATG5、ATG12及自噬标志分子LC3 mRNA表达均降低；当给予TREM-1拮抗剂后，巨噬细胞的ATG7、ATG5、ATG12、LC3 mRNA表达均升高，而采用TREM-1激动剂后，自噬基因表达被抑制。Western Blot检测结果显示，TREM-1可抑制LPS应激下巨噬细胞LC3 Ⅱ/Ⅰ、ATG7蛋白的表达。免疫荧光检测表明TREM-1激动剂可使巨噬细胞胞内的LC3蛋白表达量减少。 结论 巨噬细胞胞膜上TREM-1激活后，可使巨噬细胞自噬减弱，提示LPS应激下的巨噬细胞TREM-1可能通过抑制巨噬细胞的正常自噬从而发挥炎症放大作用。
关键词： ; 髓样细胞表达的触发受体-1; 巨噬细胞; 脂多糖; 自噬
朱爱萍 , 梁道淼 , 粟青 , 李莉芳 , 贾哲 , 贺丽萍 , 任翔 , 孙国瑛 . 髓样细胞表达的触发受体1抑制LPS应激下巨噬细胞的自噬[J]. 中国临床解剖学杂志, 2018 , 36(6) : 642 -647 . DOI: 10.13418/j.issn.1001-165x.2018.06.009
Objective To observe the effect of TREM-1 on the expression of autophagy related genes in macrophages. Methods The expression of TREM-1 protein in macrophages exposed to lipopolysaccharide (LPS) was observed. TREM-1 agonists (MAB1187) and TREM-1 antagonists (LR12) were used to regulate the activities of macrophage. qPCR was used to detect macrophage autophagy related protein ATG7, ATG5, ATG12, and microtubule associated protein 1 light chain 3 (LC3) mRNA, respectively. Expression of TREM-1, LC3 II/I, and ATG7 protein were detected by Western Blot. The expression of autophagy marker protein LC3 in macrophages under LPS stress and TREM-1 activation was detected by immunofluorescence. Results The expression of TREM-1 protein in macrophages was significantly increased under the stress of LPS (400 ng/mL, 1000 ng/mL), and the TREM-1 protein expression in macrophages reached a peak when LPS (1000 ng/mL) acted as macrophage 24 h. ATG7, ATG5, ATG12 and LC3 mRNA expression of macrophage were reduced by LPS stress. When TREM-1 antagonists were given, the expression of ATG7, ATG5, ATG12 and LC3 mRNA increased, and the autophagic gene expression of macrophage was reversed after the use of TREM-1 activator. Western Blot assay showed that TREM-1 inhibited the expression of LC3 II/I and ATG7 proteins in macrophages under LPS stress. Immunofluorescence assay showed that TREM-1 could reduce the content of LC3 protein in macrophages. Conclusions The macrophage autophagy is inhibited after the activation of TREM-1 on the membrane of macrophage, suggesting that the TREM-1 of macrophage under LPS stress may play an inflammatory magnification by inhibiting the normal autophagy of macrophages.
Key words： ; TREM-1; Macrophage; LPS; Autophagy
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