Maresin-1通过ALX/NF-κB通路促进巨噬细胞的自噬

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摘要目的：探讨促炎症消退介质maresin-1（MaR1）对巨噬细胞自噬的影响及其机制。方法：培养小鼠骨髓来源巨噬细胞，给予MaR1后，用激光扫描共聚焦显微镜技术观察LC3B聚集，用Western blot法测定LC3B与LC3A比值，用qPCR法测定ATG5、ATG7、BECN-1基因表达。用溶酶体抑制剂氯喹（CQ）、PI3K抑制剂（LY294002）、雷帕霉素、ALX抑制剂Boc-2分别预处理细胞后，测定beclin-1表达和LC3B与LC3A比值的变化。取p50-/-和WT小鼠，提取骨髓巨噬细胞，给予MaR1刺激，测定LC3B与LC3A比值的变化，及p50、p65的表达。结果：流式细胞仪测定结果显示F4/80和CD16的共表达达90%以上。巨噬细胞经MaR1作用后，LC3B的荧光强度明显增强，LC3B与LC3A的比值上升（P＜0.05），ATG5、ATG7、BECN-1基因表达量增高（P＜0.01）。经CQ或雷帕霉素预处理后，MaR1仍旧使巨噬细胞自噬增强。而加入LY294002预处理后，MaR1促进自噬的作用消失。经MaR1作用，巨噬细胞的p50表达量增多（P＜0.01），且在p50-/-小鼠提取的巨噬细胞上，MaR1促进自噬的作用消失。结论：MaR1通过ALX/NF-κB通路促进骨髓来源的巨噬细胞的自噬。

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	林静
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关键词 ：自噬；maresin-1；NF-&kappa, B；巨噬细胞；mTOR

Abstract：Objective: To detect the activation of macrophage autophgy caused by maresin-1 (MaR1) and the possible related signaling pathway. Methods: The mice bone marrow-derived macrophages (BMDMs) was cultured in vitro. The expressin of LC3B was observed under laser scanning confocal microscope, the ratio of LC3B and LC3A was detected by Western blot, the expresion of ATG5, ATG7, BECN-1 gene were detected by qPCR after giving MaR1. Then macrophages were pretreated separately with lysosomal inhibitors (CQ), PI3K inhibitors (LY294002), mTOR inhibitors (rapamycin), ALX inhibitors (Boc-2), and the ratio of LC3B and LC3A or the expression of beclin-1 were detected. Results: Mice BMDMs were consisted of approximately 90% F4/80 expression asassessed by flow cytometry. The expression of LC3B was distinctly increased when given MaR1 which observed under laser scanning confocal microscope (P<0.05), so is the ratio of LC3B and LC3A which detected by Western blot (P<0.05), and the expression of ATG5, ATG7, BECN-1 which detected by qPCR (P<0.01). When cells were pretreated by CQ or rapamycin, the effect of MaR1 was not diappeared, when pretreated by LY294002, the effect of MaR1 was diappeared. After treatment by MaR1, the expression of p50 was increased (P<0.01), the effect of MaR1 in the macropages come from p50 KO mice was disappeard. Conclusion: MaR1 activates autophagy in macrophages via ALX/NF-κB pathway.

Key words： autophagy maresin-1 NF-kappa B macrophages mTOR

收稿日期: 2017-03-01

基金资助:国家自然科学基金资助项目（81571862）。

通讯作者: 高昉，教授，博士生导师，Email：f.gaosmith@bham.ac.uk

作者简介: 林静（1991-），女，浙江温州人，硕士生。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I7/474

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