Kido Rie, Hiroshima Yuka, Kido Jun-Ichi, Bando Mika, Yoshida Kaya, Inagaki Yuji, Yumoto Hiromichi
Department of Periodontology and Endodontology, Tokushima University Graduate School of Biomedical Sciences, Japan.
Department of Oral Microbiology, Tokushima University Graduate School of Biomedical Sciences, Japan.
J Oral Biosci. 2025 Mar;67(1):100624. doi: 10.1016/j.job.2025.100624. Epub 2025 Feb 1.
Lipocalin 2 (LCN2) is a glycoprotein with multiple functions, including antimicrobial activity, inflammatory response modulation, and cell migration. LCN2 is expressed in some cells, such as epithelial cells and neutrophils, and its levels are increased in inflammatory diseases. This study investigated the presence of LCN2 receptor (24p3R) in cells around periodontal tissues and function of LCN2 in cells with a receptor to explore the role of LCN2 in periodontal diseases.
The presence of 24p3R was examined in periodontal cells, including human gingival fibroblasts, periodontal ligament fibroblasts, human oral epithelial cells (HOECs), and neutrophil-like cells (HL-60), by Western blotting. Changes in periodontal disease-associated proteins in the presence of recombinant LCN2 (rLCN2) were examined using a protein array in differentiated HL-60 (dHL-60) cells. Interleukin-8 (IL-8) and macrophage inflammatory protein-1α (MIP-1α) mRNA expressions were analyzed by qRT-PCR, and IL-8 and MIP-1α levels in dHL-60 cells treated with rLCN2 or Porphyromonas gingivalis-lipopolysaccaharide (P.g-LPS) were determined by enzyme-linked immunosorbent assay.
We detected 24p3R in dHL-60 cells. IL-8 was highly expressed and MIP-1α was weakly expressed in dHL-60 cells using a protein array. rLCN2 significantly decreased IL-8 mRNA and protein levels and suppressed P.g-LPS-induced IL-8 production in dHL-60 cells. As dHL-60 cells were co-cultured with HOECs in which LCN2 was knocked down, IL-8 mRNA expression increased in dHL-60 cells. Furthermore, rLCN2 inhibited MIP-1α production in dHL-60 cells.
LCN2 suppresses inflammatory responses by regulating IL-8 and MIP-1α expression in periodontal diseases.
脂质运载蛋白2（LCN2）是一种具有多种功能的糖蛋白，包括抗菌活性、炎症反应调节和细胞迁移。LCN2在某些细胞中表达，如上皮细胞和中性粒细胞，并且其水平在炎症性疾病中升高。本研究调查了牙周组织周围细胞中LCN2受体（24p3R）的存在情况以及LCN2在有该受体的细胞中的功能，以探讨LCN2在牙周疾病中的作用。
通过蛋白质印迹法检测牙周细胞中24p3R的存在情况，这些牙周细胞包括人牙龈成纤维细胞、牙周膜成纤维细胞、人口腔上皮细胞（HOECs）和中性粒细胞样细胞（HL-60）。使用蛋白质芯片检测在重组LCN2（rLCN2）存在的情况下牙周疾病相关蛋白的变化，该蛋白质芯片用于分化的HL-60（dHL-60）细胞。通过qRT-PCR分析白细胞介素-8（IL-8）和巨噬细胞炎性蛋白-1α（MIP-1α）的mRNA表达，并通过酶联免疫吸附测定法测定用rLCN2或牙龈卟啉单胞菌脂多糖（P.g-LPS）处理的dHL-60细胞中IL-8和MIP-1α的水平。
我们在dHL-60细胞中检测到了24p3R。使用蛋白质芯片发现IL-8在dHL-60细胞中高表达，而MIP-1α在dHL-60细胞中弱表达。rLCN2显著降低dHL-60细胞中IL-8的mRNA和蛋白水平，并抑制P.g-LPS诱导的dHL-60细胞中IL-8的产生。当dHL-60细胞与LCN2被敲低的HOECs共培养时，dHL-60细胞中IL-8的mRNA表达增加。此外，rLCN2抑制dHL-60细胞中MIP-1α的产生。
LCN2通过调节牙周疾病中IL-8和MIP-1α的表达来抑制炎症反应。
