A20 suppresses vascular inflammation by recruiting proinflammatory signaling molecules to intracellular aggresomes
A20 suppresses vascular inflammation by recruiting proinflammatory signaling molecules to intracellular aggresomes
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DOI：
10.1096/fj.14-258533
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发表时间：
2015-05-01
期刊：
影响因子：
4.8
通讯作者：
Evans, Paul C.
中科院分区：
文献类型：
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作者：
Enesa, Karine;Moll, Herwig P.;Evans, Paul C.
A20通过抑制炎症转录因子NF-kappa b来防止病理性血管重构。A20的功能归因于受体相互作用蛋白1 （RIP1）的泛素编辑，从而影响其活性/稳定性。用移植血管病变和人类细胞的小鼠模型测试了这一机制的有效性。用含有A20（或β -半乳糖苷酶作为对照）的腺病毒转导小鼠C57BL/6主动脉，移植到主要组织相容性复合物不匹配的BALB/c小鼠体内。将原代内皮细胞、平滑肌细胞或转化上皮细胞（均为人类）转染野生型A20或催化失活突变体作为对照。通过报告基因检测、免疫荧光染色和Western blotting检测NF-kappa B活性和RIP1的细胞内定位。原生A20和催化失活A20对NF-kappa B活性的抑制作用相似（-70% vs -76%; P < 0.05）。在小鼠异体血管移植物和人细胞中，A20促进RIP1向不溶性聚合体的定位（53%对0%），而不改变RIP1的表达，多泛素链的组装增加了这一过程（87%对28%，P < 0.05）。A20捕获不溶性聚合体中的多泛素化信号中间体，从而降低其下游nf - κ B信号的生物利用度。这一新机制有助于保护外源性A20治疗的移植器官免于血管病变。
A20 protects against pathologic vascular remodeling by inhibiting the inflammatory transcription factor NF-kappa B. A20's function has been attributed to ubiquitin editing of receptor-interacting protein 1 (RIP1) to influence activity/stability. The validity of this mechanism was tested using a murine model of transplant vasculopathy and human cells. Mouse C57BL/6 aortae transduced with adenoviruses containing A20 (or beta-galactosidase as a control) were allografted into major histocompatibility complex-mismatched BALB/c mice. Primary endothelial cells, smooth muscle cells, or transformed epithelial cells (all human) were transfected with wild-type A20 or with catalytically inactive mutants as a control. NF-kappa B activity and intracellular localization of RIP1 was monitored by reporter gene assay, immunofluorescent staining, and Western blotting. Native and catalytically inactive versions of A20 had similar inhibitory effects on NF-kappa B activity (-70% vs. -76%; P > 0.05). A20 promoted localization of RIP1 to insoluble aggresomes in murine vascular allografts and in human cells (53% vs. 0%) without altering RIP1 expression, and this process was increased by the assembly of polyubiquitin chains (87% vs. 28%; P < 0.05). A20 captures polyubiquitinated signaling intermediaries in insoluble aggresomes, thus reducing their bioavailability for downstream NF-kappa B signaling. This novel mechanism contributes to protection from vasculopathy in transplanted organs treated with exogenous A20.