Niu Ya, Xu Guangyu, Zhu Shaoping, Wei Xiurong, Wu Changli, Zhang Ruigang, Chen Chunling, Yan Lvbin, Luo Haihua, Deng Simin, Wu Weijian, Li Yaojing, Liu Ming, Jiang Yong, Zhang Xiujuan
Department of Physiology, Guangdong Medical University, Zhanjiang, Guangdong 524023, China.
Laboratory Animal Center, Guangdong Medical University, Zhanjiang, Guangdong 524023, China.
Mol Immunol. 2023 Jan;153:94-105. doi: 10.1016/j.molimm.2022.11.017. Epub 2022 Nov 29.
The massive release of pro-inflammatory cytokines is a crucial step in triggering the inflammatory cascade in sepsis. Exploring the key molecules regulating the expression and release of multiple cytokines has important value for revealing the mechanism of the cytokine storm in sepsis. This study aimed to investigate the role of multifunctional nuclear protein non-POU domain containing octamer-binding protein (NONO) in the sepsis cytokine storm and to elucidate the underlying mechanism. We found that NONO expression in tissues and cells of sepsis mice was significantly upregulated. Downregulation of NONO expression inhibited the mRNA expression of multiple cytokines, including IL-6, IL-1β, MCP-1, MIP-1α, and MIP-1β in inflammatory cells from mice and human leukemic monocyte-THP1 cells challenged with lipopolysaccharide (LPS), and significantly decreased the level of these cytokines and TNF-α in the supernatant of THP1 cells challenged by LPS. Nono knockout also reduced the levels of TNF-α, IL-6, MIP-1α, and MIP-1β in serum, alleviated hepatocyte edema, and improved the survival rate of sepsis mice. Reduced NONO expression decreased the phospho-ERK1/2 level in inflammatory cells from sepsis mice or THP1 cells challenged by LPS. Phospho-ERK1/2 inhibitor decreased the mRNA expression and concentration of cytokines in the culture supernatant of LPS-induced THP1 cells, similar to the effect of NONO knockdown. After LPS challenge, the levels of phospho-ERK1/2 and NONO were increased, with obvious colocalization in the nucleus and vesicular-like organelles in macrophages. NONO knockdown decreased nuclear translocation of phospho-ERK1/2 in LPS-challenged THP1 cells. These results suggest that NONO is a potentially critical molecule involved in multiple cytokine production in sepsis. Upregulated NONO in sepsis may promote the expression and release of multiple cytokines to participate in a sepsis cytokine storm by promoting ERK1/2 phosphorylation.
促炎细胞因子的大量释放是引发脓毒症炎症级联反应的关键步骤。探索调节多种细胞因子表达和释放的关键分子对于揭示脓毒症细胞因子风暴的机制具有重要价值。本研究旨在探讨多功能核蛋白含八聚体结合蛋白的非POU结构域（NONO）在脓毒症细胞因子风暴中的作用，并阐明其潜在机制。我们发现脓毒症小鼠组织和细胞中NONO表达显著上调。下调NONO表达可抑制多种细胞因子的mRNA表达，包括脂多糖（LPS）刺激的小鼠炎症细胞和人白血病单核细胞-THP1细胞中的白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、单核细胞趋化蛋白-1（MCP-1）、巨噬细胞炎性蛋白-1α（MIP-1α）和巨噬细胞炎性蛋白-1β（MIP-1β），并显著降低LPS刺激的THP1细胞上清液中这些细胞因子和肿瘤坏死因子-α（TNF-α）的水平。敲除Nono还可降低血清中TNF-α、IL-6、MIP-1α和MIP-1β的水平，减轻肝细胞水肿，并提高脓毒症小鼠的存活率。NONO表达降低可降低脓毒症小鼠炎症细胞或LPS刺激的THP1细胞中磷酸化细胞外信号调节激酶1/2（phospho-ERK1/2）的水平。磷酸化ERK1/2抑制剂可降低LPS诱导的THP1细胞培养上清液中细胞因子的mRNA表达和浓度，类似于敲低NONO的效果。LPS刺激后，磷酸化ERK1/2和NONO水平升高，在巨噬细胞核和囊泡样细胞器中有明显共定位。敲低NONO可减少LPS刺激的THP1细胞中磷酸化ERK1/2的核转位。这些结果表明，NONO是脓毒症中参与多种细胞因子产生的潜在关键分子。脓毒症中NONO上调可能通过促进ERK1/2磷酸化来促进多种细胞因子的表达和释放，从而参与脓毒症细胞因子风暴。
