Augmenting ATG14 alleviates atherosclerosis and inhibits inflammation via promotion of autophagosome-lysosome fusion in macrophages
Augmenting ATG14 alleviates atherosclerosis and inhibits inflammation via promotion of autophagosome-lysosome fusion in macrophages
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DOI：
10.1080/15548627.2021.1909833
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发表时间：
2021-04
期刊：
影响因子：
13.3
通讯作者：
Hui Zhang;S. Ge;Buqing Ni;Keshuai He;Pengcheng Zhu;Xiaohong Wu;Y. Shao
中科院分区：
文献类型：
--
作者：
Hui Zhang;S. Ge;Buqing Ni;Keshuai He;Pengcheng Zhu;Xiaohong Wu;Y. Shao
巨噬细胞中的巨自噬/自噬功能障碍有助于动脉粥样硬化。自噬-溶酶体降解系统受损导致脂质蓄积，促进动脉粥样硬化斑块形成。ATG 14是自噬体与溶酶体融合的重要调节因子。ATG 14是否在动脉粥样硬化中的巨噬细胞自噬功能障碍中起作用尚不清楚。为了研究ATG 14对巨噬细胞自噬的影响，评价了人动脉粥样硬化斑块、apoe-/-小鼠和培养的小鼠巨噬细胞。利用腺病毒过表达ATG 14来揭示其在自噬、炎症和动脉粥样硬化斑块形成中的功能。结果表明，人和小鼠动脉粥样硬化斑块中巨噬细胞存在自噬功能受损，ATG 14表达减少。Ox-LDL损害巨噬细胞中自噬体-溶酶体融合，降低ATG 14表达。ATG 14在巨噬细胞中的过表达增强了自噬体与溶酶体的融合，促进了脂质降解，减少了Ox-LDL诱导的细胞凋亡和炎症反应。增强ATG 14表达逆转了apoe−/-小鼠斑块巨噬细胞中的自噬功能障碍，减弱了SQSTM 1/p62的积累，抑制了炎症，并上调了Treg细胞的数量，从而减轻了动脉粥样硬化病变。缩略语：ABCC1：ATP结合盒，亚家族C（CFTR/MRP），成员1; ABCA 1：ATP结合盒，亚家族A（ABC 1），成员1; Ad-Atg 14：携带小鼠Atg 14基因的腺病毒载体; Ad-LacZ：携带细菌β-半乳糖苷酶基因的腺病毒载体; apoe-/-：载脂蛋白E敲除; ATG 14：自噬相关14; CD 68：CD 68抗原; DAPI：4 β，6-二脒基-2-苯基吲哚; Dil-ox-LDL：Dil-氧化低密度脂蛋白; ELISA：酶联免疫吸附试验; HFD：高脂饮食（致动脉粥样硬化饮食）; IL：白细胞介素; LAMP 2：溶酶体相关膜蛋白2; LDL-C：低密度脂蛋白胆固醇; MAP 1 LC 3/LC 3：微管相关蛋白1轻链3; ND：正常饮食; Ox-LDL：氧化低密度脂蛋白; PBMC：外周血单核细胞; SQSTM 1/p62：隔离体1; SREBF 1/SREBP 1c：固醇调节元件结合转录因子1; SREBF 2/SREBP 2：固醇调节元件结合因子2; STX 17：突触融合蛋白17; TC：血清总胆固醇; TG：甘油三酯; TNF：肿瘤坏死因子; IFN：干扰素; Treg细胞：调节性T细胞
ABSTRACT Dysfunction of macroautophagy/autophagy in macrophages contributes to atherosclerosis. Impaired autophagy-lysosomal degradation system leads to lipid accumulation, facilitating atherosclerotic plaque. ATG14 is an essential regulator for the fusion of autophagosomes with lysosomes. Whether ATG14 plays a role in macrophage autophagy dysfunction in atherosclerosis is unknown. To investigate the effects of ATG14 on macrophage autophagy, human atherosclerotic plaque, apoe−/- mice and cultured mouse macrophages were evaluated. Overexpression of ATG14 by adenovirus was used to reveal its function in autophagy, inflammation and atherosclerotic plaque formation. Results showed that impaired autophagy function with reduction of ATG14 expression existed in macrophages of human and mouse atherosclerotic plaques. Ox-LDL impaired autophagosome-lysosome fusion with reduction of ATG14 expression in macrophages. Overexpression of ATG14 in macrophages enhanced fusion of autophagosomes with lysosomes and promoted lipid degradation, decreasing Ox-LDL-induced apoptosis and inflammatory response. Augmenting ATG14 expression reversed the autophagy dysfunction in macrophages of apoe−/- mice plaque, blunted SQSTM1/p62 accumulation, inhibited inflammation, and upregulated the population of Treg cells, resulting in alleviating atherosclerotic lesions. Abbreviations: ABCC1: ATP-binding cassette, sub-family C (CFTR/MRP), member 1; ABCA1: ATP-binding cassette, sub-family A (ABC1), member 1; Ad-Atg14: adenovirus vector carrying the mouse Atg14 gene; Ad-LacZ: adenovirus vector carrying the gene for bacterial β-galactosidase; apoe−/- : apolipoprotein E knockout; ATG14: autophagy-related 14; CD68: CD68 antigen; DAPI: 4ʹ,6-diamidino-2-phenylindole; Dil-ox-LDL: Dil-oxidized low density lipoprotein; ELISA: enzyme-linked immunosorbent assay; HFD: high-fat diet (an atherogenic diet); IL: interleukin; LAMP2: lysosomal-associated membrane protein 2; LDL-C: low density lipoprotrein cholesterol; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; ND: normal diet; Ox-LDL: oxidized low density lipoprotein; PBMC: peripheral blood mononuclear cells; SQSTM1/p62: sequestosome 1; SREBF1/SREBP1c: sterol regulatory element binding transcription factor 1; SREBF2/SREBP2: sterol regulatory element binding factor 2; STX17: syntaxin 17; TC: serum total cholesterol; TG: triglyceride; TNF: tumor necrosis factor; IFN: interferon; Treg cell: regulatory T cell