自噬通过下调ATF6通路抑制促炎型巨噬细胞活化  

Autophagy inhibits activation of pro-inflammatory macrophages by suppressing ATF6 pathway

作　　者：李慧媛 黄小荣 黄丽凤 杨陈 安宁 LI Huiyuan;HUANG Xiaorong;HUANG Lifeng;YANG Chen;AN Ning(Guangdong Provincial Key Laboratory of Autophagy and Major Chronic Non-communicable Diseases,Institute of Nephrology,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001,China)

机构地区：[1]广东医科大学附属医院肾病研究所,省市共建细胞自噬与慢性重大非传染性疾病研究广东省重点实验室,湛江524001

出　　处：《中国免疫学杂志》2025年第4期861-866,共6页Chinese Journal of Immunology

基　　金：广东省基础与应用基础研究基金(2023A1515030024,2021A1515011581);省市共建细胞自噬与慢性重大非传染性疾病研究广东省重点实验室项目(2022B1212030003);国家临床重点专科建设(肾病科)单位项目。

摘　　要：目的:揭示自噬抑制促炎型巨噬细胞过度活化的可能分子机制。方法:利用LysM-Cre小鼠和Atg5^(flox/+)小鼠繁殖巨噬细胞自噬缺陷小鼠(Atg5^(△mye))。使用巨噬细胞集落刺激因子诱导小鼠骨髓细胞分化为骨髓源性巨噬细胞(BMDMs)。使用不同浓度的脂多糖(LPS)刺激BMDMs,利用Western blot检测BMDMs内葡萄糖调节蛋白78(GRP78)等内质网应激分子、iNOS等炎症因子以及LC3-Ⅱ和p62等自噬相关蛋白的表达变化。利用LPS刺激来自Atg5^(flox/flox)和Atg5^(△mye)小鼠的BMDMs,并进行蛋白组学检测。利用牛磺熊去氧胆酸(TUDCA)或激活转录因子6(ATF6)抑制剂Ceapin-A7处理BMDMs。结果:LPS促进内质网应激相关蛋白表达和促炎型巨噬细胞活化,同时抑制BMDMs自噬。通过蛋白组学检测和GO富集分析发现,BMDMs自噬缺陷引起内质网应激关键下游ATF6通路变化。利用TUDCA抑制内质网应激,则显著下调自噬缺陷引起的BMDMs内iNOS的表达以及炎症因子IL-18和IL-1β分泌。利用ATF6特异性抑制剂(Ceapin-A7)处理LPS刺激下的BMDMs,发现Ceapin-A7显著下调Atg5缺失引起的iNOS和炎症因子表达增加。结论:自噬通过下调ATF6通路抑制促炎型巨噬细胞活化。Objective:To explore the potential mechanisms of autophagy suppressing the activation of pro-inflammatory macrophages.Methods:Macrophage-specific knockout mice(Atg5^(△mye))was generated by the hybridization of Lyz2-Cre mice and Atg 5^(flox/+)mice.Bone marrow cells were differentiated into bone marrow-derived macrophages(BMDMs)induced by monocyte colony-stimulating factor.The expressions of endoplasmic reticulum stress(ER stress)-related proteins,such as glucose regulatory protein 78(GRP78),autophagy-related marker LC3-Ⅱand p62,and pro-inflammatory iNOS,were detected by Western blot.GO analysis was performed on BMDMs from Atg5^(flox/flox) and Atg5^(△mye) mice stimulated by LPS.Tauroursodeoxycholate(TUDCA)or ATF6 inhibitor Ceapin-A7 was used to suppress the ER stress in BMDMs.Results:LPS promoted the expression of ER stress-related proteins and the activation of pro-inflammatory macrophages,while inhibiting autophagy in BMDMs.By utilizing proteomic detection and GO enrichment analysis,it was found that the autophagy deficiency in BMDMs caused changes in the ATF6 pathway,a key downstream pathway of ER stress.Inhibition of ER stress by TUDCA significantly down-regulated the expression of iNOS and the secretion of inflammatory cytokines IL-18 and IL-1βin autophagy-deficient BMDMs.ATF6-specific inhibitor(Ceapin-A7)was used to exculpate LPS-stimulated BMDMs,and it was found that Ceapin-A7 significantly down-regulated the elevated expression of iNOS and inflammatory factors caused by Atg5 deletion in BMDMs.Conclusion:Autophagy inhibits the activation of pro-inflammatory macrophages by suppressing ATF6 pathway.

关 键 词：巨噬细胞 自噬 内质网应激 炎症反应 

分 类 号：R392.12[医药卫生—免疫学]