Yu Yaqiong, Yang Di, Qiu Lihong, Okamura Hirohiko, Guo Jiajie, Haneji Tatsuji
Department of Endodontics, School of Stomatology, China Medical University, Shenyang, Liaoning 110002, P.R. China.
Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Kuramoto, Tokushima 770-8504, Japan.
Mol Med Rep. 2014 Sep;10(3):1371-6. doi: 10.3892/mmr.2014.2353. Epub 2014 Jun 25.
Osteoblasts produce various types of cytokines under pathological conditions and control osteoclast differentiation. Tumor necrosis factor-α (TNF-α) has been demonstrated to exert complex effects in osteoblasts under local inflammatory conditions, including in periodontal and periapical diseases. Interleukin-34 (IL-34) has been recently identified as a novel regulatory factor for the differentiation and function of osteoclasts. The present study provides the first evidence, to the best of our knowledge, that the expression of IL-34 is induced by TNF-α through nuclear factor-κB (NF-κB) activation in MC3T3-E1 osteoblastic cells. TNF-α induced IL-34 expression in a dose- and time-dependent manner. Immunocytochemistry with an NF-κB antibody demonstrated that NF-κB was mainly localized in the cytoplasm of the untreated MC3T3-E1 cells. Rapid translocation of NF-κB from the cytoplasm to the nucleus was observed in the cells treated with TNF-α for 15 min. Translocation and transcriptional activity of NF-κB were also determined by western blotting and a luciferase reporter assay, respectively. Pretreatment with 100 µM CAPE, an inhibitor of NF-κB, significantly inhibited TNF-α-induced IL-34 expression. These results indicate that TNF-α induces IL-34 expression via NF-κB in osteoblasts.
在病理条件下,成骨细胞会产生多种细胞因子并控制破骨细胞的分化。肿瘤坏死因子-α(TNF-α)已被证明在局部炎症条件下,包括在牙周病和根尖周病中,对成骨细胞发挥复杂的作用。白细胞介素-34(IL-34)最近被确定为破骨细胞分化和功能的一种新型调节因子。据我们所知,本研究首次提供证据表明,在MC3T3-E1成骨细胞中,TNF-α通过激活核因子-κB(NF-κB)诱导IL-34的表达。TNF-α以剂量和时间依赖性方式诱导IL-34表达。用NF-κB抗体进行免疫细胞化学分析表明,NF-κB主要定位于未处理的MC3T3-E1细胞的细胞质中。在用TNF-α处理15分钟的细胞中,观察到NF-κB从细胞质快速转移到细胞核。分别通过蛋白质印迹法和荧光素酶报告基因检测法测定NF-κB的转移和转录活性。用100μM的NF-κB抑制剂CAPE预处理可显著抑制TNF-α诱导的IL-34表达。这些结果表明,TNF-α通过NF-κB在成骨细胞中诱导IL-34表达。