Moore Tyler C, Pinkerton Terrence Scott, Petro Thomas M
Department of Biology, College of Science and Technology, Bellevue University, Bellevue, NE, 68005, USA.
Department of Oral Biology, University of Nebraska Medical Center, Lincoln, NE, 68583, USA.
J Inflamm Res. 2025 Jan 27;18:1255-1265. doi: 10.2147/JIR.S496930. eCollection 2025.
Macrophage responses to lipopolysaccharides (LPS) drive inflammatory diseases, such as periodontitis, with production of IL-6 and Nitric Oxide (NO). However, anti-inflammatory macrophages counter inflammation with the production of CCL22. Interferon regulatory factor 3 (IRF3) plays a significant role in expression of both IL-6 and NO during macrophage responses through Interferon-stimulated Response Elements (ISREs) of promoters.
To determine the role of IRF3 in LPS-induced pro- and anti-inflammatory macrophage responses, we used the macrophage cell line RAW264.7 modified with an ISRE promoter driving secreted luciferase (RAW264.7-Lucia) to assess IRF3 activity in response to and LPS. For comparison, responses to poly I:C and IFN-gamma and responses from RAW264.7 cells deficient in IRF3 were also assessed.
Herein, LPS of , significantly enhanced production of IL-6 and NO that was induced by LPS but significantly decreased poly I:C-induced ISRE promoter activity. Moreover, IRF3 deficiency depressed the LPS-induced ISRE promoter activity and NO production but increased IL-6 and CCL22 in response to LPS. Restoration of IRF3 expression in IRF3KO RAW cells increased IL-6, restored NO, and decreased CCL22 production in response to LPS of .
Therefore, IRF3 is critical to the expression of pro- and anti-inflammatory factors produced by macrophages responding to LPS and could be a target during periodontitis treatment.
巨噬细胞对脂多糖（LPS）的反应会引发炎症性疾病，如牙周炎，同时产生白细胞介素-6（IL-6）和一氧化氮（NO）。然而，抗炎性巨噬细胞通过产生CCL22来对抗炎症。干扰素调节因子3（IRF3）在巨噬细胞反应过程中，通过启动子的干扰素刺激反应元件（ISREs），在IL-6和NO的表达中发挥重要作用。
为了确定IRF3在LPS诱导的促炎和抗炎巨噬细胞反应中的作用，我们使用了用驱动分泌型荧光素酶的ISRE启动子修饰的巨噬细胞系RAW264.7（RAW264.7-Lucia）来评估对不同浓度LPS反应时的IRF3活性。为作比较，还评估了对聚肌胞苷酸（poly I:C）和干扰素-γ（IFN-γ）的反应以及IRF3缺陷的RAW264.7细胞的反应。
在此，1μg/mL的LPS显著增强了由0.1μg/mL LPS诱导的IL-6和NO的产生，但显著降低了聚肌胞苷酸诱导的ISRE启动子活性。此外，IRF3缺陷降低了LPS诱导的ISRE启动子活性和NO产生，但增加了对LPS反应时的IL-6和CCL22。在IRF3基因敲除（IRF3KO）的RAW细胞中恢复IRF3表达，增加了对1μg/mL LPS反应时的IL-6，恢复了NO，并降低了CCL22的产生。
因此，IRF3对于巨噬细胞对LPS反应所产生的促炎和抗炎因子的表达至关重要，并且可能是牙周炎治疗期间的一个靶点。
