Department of Anesthesiology, University of Illinois College of Medicine, Chicago, Illinois, United States.
Department of Pharmacology and Regenerative Medicine, University of Illinois College of Medicine, Chicago, Illinois, United States.
Am J Physiol Lung Cell Mol Physiol. 2023 May 1;324(5):L596-L608. doi: 10.1152/ajplung.00328.2022. Epub 2023 Mar 7.
Inflammasome activation is of central importance for the process of generation of overwhelming inflammatory response and the pathogenesis of sepsis. The intrinsic molecular mechanism for controlling inflammasome activation is still poorly understood. Here we investigated the role of p120-catenin expression in macrophages in regulating nucleotide-binding oligomerization domain (NOD) and leucine-rich repeat (LRR)- and pyrin domain-containing proteins 3 (NLRP3) inflammasome activation. Depletion of p120-catenin in murine bone marrow-derived macrophages enhanced caspase-1 activation and secretion of active interleukin (IL)-1β in response to ATP stimulation following LPS priming. Coimmunoprecipitation analysis showed that p120-catenin deletion promoted NLRP3 inflammasome activation by accelerating the assembly of the inflammasome complex comprised of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and pro-caspase-1. Depletion of p120-catenin also increased the production of mitochondrial reactive oxygen species. Pharmacological inhibition of mitochondrial reactive oxygen species nearly completely abolished NLRP3 inflammasome activation, caspase-1 activation, and the production of IL-1β in p120-catenin-depleted macrophages. Furthermore, p120-catenin ablation significantly disrupted mitochondrial function, evidenced by decreased mitochondrial membrane potential and lower production of intracellular ATP. In alveolar macrophage-depleted mice challenged with cecal ligation and puncture, pulmonary transplantation of p120-catenin-deficient macrophages dramatically enhanced the accumulation of IL-1β and IL-18 in bronchoalveolar lavage fluid. These results demonstrate that p120-catenin prevents NLRP3 inflammasome activation in macrophages by maintaining mitochondrial homeostasis and reducing the production of mitochondrial reactive oxygen species in response to endotoxin insult. Thus, inhibition of NLRP3 inflammasome activation by stabilization of p120-catenin expression in macrophages may be a novel strategy to prevent an uncontrolled inflammatory response in sepsis.
炎症小体的激活对于产生过度炎症反应和脓毒症发病机制的过程至关重要。控制炎症小体激活的内在分子机制仍知之甚少。在这里,我们研究了巨噬细胞中 p120-连环蛋白表达在调节核苷酸结合寡聚结构域 (NOD) 和富含亮氨酸重复 (LRR) 和吡喃结构域蛋白 3 (NLRP3) 炎症小体激活中的作用。在 LPS 引发后,用 ATP 刺激耗尽小鼠骨髓来源的巨噬细胞中的 p120-连环蛋白会增强半胱天冬酶-1 的激活和活性白细胞介素 (IL)-1β的分泌。共免疫沉淀分析表明,p120-连环蛋白缺失通过加速由 NLRP3、凋亡相关斑点样蛋白含有半胱氨酸酶募集域 (ASC) 和前半胱天冬酶-1 组成的炎症小体复合物的组装来促进 NLRP3 炎症小体的激活。p120-连环蛋白的缺失也增加了线粒体活性氧的产生。线粒体活性氧的药理学抑制几乎完全消除了 p120-连环蛋白耗尽的巨噬细胞中的 NLRP3 炎症小体激活、半胱天冬酶-1 的激活和 IL-1β的产生。此外,p120-连环蛋白缺失显着破坏了线粒体功能,表现为线粒体膜电位降低和细胞内 ATP 产量降低。在盲肠结扎和穿刺术挑战的肺泡巨噬细胞耗尽的小鼠中,p120-连环蛋白缺陷型巨噬细胞的肺移植显着增加了支气管肺泡灌洗液中 IL-1β和 IL-18 的积累。这些结果表明,p120-连环蛋白通过维持线粒体稳态和减少内毒素损伤后线粒体活性氧的产生来防止巨噬细胞中 NLRP3 炎症小体的激活。因此,通过稳定巨噬细胞中 p120-连环蛋白的表达来抑制 NLRP3 炎症小体的激活可能是预防脓毒症中失控炎症反应的一种新策略。