Hydrogen peroxide-induced oxidative stress promotes expression of CXCL15/Lungkine mRNA in a MEK/ERK-dependent manner in fibroblast-like synoviocytes derived from mouse temporomandibular joint
Hydrogen peroxide-induced oxidative stress promotes expression of CXCL15/Lungkine mRNA in a MEK/ERK-dependent manner in fibroblast-like synoviocytes derived from mouse temporomandibular joint
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DOI：
10.1016/j.job.2022.12.002
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发表时间：
2023-03-02
影响因子：
2.4
通讯作者：
Ishisaki, Akira
中科院分区：
文献类型：
--
作者：
Asanuma, Kanna;Yokota, Seiji;Ishisaki, Akira
目的：颞下颌关节骨关节炎（TMJ-OA）是一种由炎症和氧化应激引起的多因素疾病。据推测，机械应力诱导的TMJ组织损伤诱导活性氧（ROS）的产生，例如羟基自由基（OH），在滑液（SF）中。一般来说，ROS的过度产生有助于OA中软骨下骨的滑膜炎症和功能障碍。然而，ROS损伤的滑膜细胞募集炎症细胞到TMJ-OA lesions.Methods的机制尚不清楚：逆转录-定量聚合酶链反应（RT-qPCR）进行评估的mRNA表达的化学引诱物分子。使用western blot analysis.Results：过氧化氢（H2O2）处理显着促进中性粒细胞趋化因子CXCL15/Lungkine的mRNA表达的成纤维细胞样滑膜细胞（FLS）来自小鼠TMJ的剂量依赖性方式（100 - 500 μ M）的细胞内信号分子的磷酸化水平进行了评估。H2O2（500 μ M）显着上调FLS细胞外信号调节激酶（ERK）1和ERK 2的磷酸化。有趣的是，丝裂原活化蛋白（MAP）/ERK激酶（MEK）抑制剂U0126（10 μ M）无效H2O2诱导的CXCL 15/Lungkine mRNA表达的增加。此外，H2O2（500 μ M）管理显着上调OH。FLS中的生产，如通过针对OH的活细胞渗透荧光探针所评估的。在荧光显微镜下。结论：H2O2诱导的氧化应激以MEK/ERK依赖的方式促进TMJ源性FLS中CXCL15/Lungkine mRNA的表达，提示FLS通过产生CXCL15/Lungkine募集中性粒细胞至TMJOA损伤区，加重局部炎症反应。(c)2022日本口腔生物学协会。由爱思唯尔公司出版。这是一篇开放获取的文章，获得了CC BY-NC-ND许可证（http：//creativecommons.org/licenses/by-nc-nd/4.0/）。
Objectives: Temporomandibular joint osteoarthritis (TMJ-OA) is a multifactorial disease caused by inflammation and oxidative stress. It has been hypothesized that mechanical stress-induced injury of TMJ tissues induces the generation of reactive oxygen species (ROS), such as hydroxyl radical (OH.), in the synovial fluid (SF). In general, the overproduction of ROS contributes to synovial inflammation and dysfunction of the subchondral bone in OA. However, the mechanism by which ROS-injured synoviocytes recruit inflammatory cells to TMJ-OA lesions remains unclear.Methods: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the mRNA expression of chemoattractant molecules. The phosphorylation levels of intracellular signaling molecules were evaluated using western blot analysis.Results: Hydrogen peroxide (H2O2) treatment significantly promoted mRNA expression of neutrophil chemoattractant CXCL15/Lungkine in a dose-dependent manner (100-500 mu M) in fibroblast-like synoviocytes (FLSs) derived from mouse TMJ. H2O2 (500 mu M) significantly upregulated the phosphorylation of extracellular signal-regulated kinase (ERK)1 and ERK2 in FLSs. Intriguingly, the mitogen-activated protein (MAP)/ERK kinase (MEK) inhibitor U0126 (10 mu M) nullified H2O2-induced increase in CXCL15/ Lungkine mRNA expression. Additionally, H2O2 (500 mu M) administration significantly upregulated OH. production in FLSs, as assessed by live-cell permeant fluorescent probe targeted against OH. under fluorescence microscopy. Furthermore, the ROS inhibitor N-acetyl-L-cysteine (5 mM) partially but significantly reversed H2O2-mediated phosphorylation of ERK1/2.Conclusions: H2O2-induced oxidative stress promoted the expression of CXCL15/Lungkine mRNA in a MEK/ERK-dependent manner in mouse TMJ-derived FLSs, suggesting that FLSs recruit neutrophils to TMJOA lesions through the production of CXCL15/Lungkine and exacerbate the local inflammatory response. (c) 2022 Japanese Association for Oral Biology. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).