Tanaka Atsuko, Hayano Satoru, Nagata Masayo, Kosami Takahiro, Wang Ziyi, Kamioka Hiroshi
Department of Orthodontics, Okayama University Hospital, Okayama, Japan.
Department of Orthodontics, Okayama University Hospital, Okayama, Japan.
Arch Oral Biol. 2023 Nov;155:105797. doi: 10.1016/j.archoralbio.2023.105797. Epub 2023 Aug 22.
This study aimed to evaluate the role of ruxolitinib in the interferon beta (IFN-β) mediated osteoblast differentiation using human dental pulp stem cells (hDPSCs).
hDPSCs from five deciduous teeth of healthy patients were stimulated by adding human recombinant IFN-β protein (1 or 2 ng/ml) to the osteogenic differentiation induction medium. Substrate formation was determined using Alizarin Red staining, calcium concentration, and osteoblast marker expression levels. Ruxolitinib was used to inhibit the Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathway. Apoptosis was detected using terminal deoxynucleotidyl nick-end labeling (TUNEL) staining, and necroptosis was detected using propidium iodide staining and phosphorylated mixed lineage kinase domain-like protein (pMLKL) expression.
In the IFN-β-treated group, substrate formation was inhibited by a reduction in alkaline phosphatase (ALP) expression in a concentration-dependent manner. Although the proliferation potency was unchanged between the IFN-β-treated and control groups, the cell number was significantly reduced in the experimental group. TUNEL-positive cell number was not significantly different; however, the protein level of necroptosis markers, interleukin-6 (IL-6) and pMLKL were significantly increased in the substrate formation. Cell number and ALP expression level were improved in the group administered ruxolitinib, a JAK-STAT inhibitor. Additionally, ruxolitinib significantly suppressed IL-6 and pMLKL levels.
Ruxolitinib interfered with the IFN-β-mediated necroptosis and osteogenic differentiation via the JAK-STAT pathway.
本研究旨在利用人牙髓干细胞（hDPSCs）评估鲁索替尼在干扰素β（IFN-β）介导的成骨细胞分化中的作用。
将人重组IFN-β蛋白（1或2 ng/ml）添加到成骨分化诱导培养基中，刺激来自健康患者五颗乳牙的hDPSCs。使用茜素红染色、钙浓度和成骨细胞标志物表达水平来确定基质形成。使用鲁索替尼抑制Janus激酶/信号转导子和转录激活子（JAK-STAT）途径。使用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色检测细胞凋亡，使用碘化丙啶染色和磷酸化混合谱系激酶结构域样蛋白（pMLKL）表达检测坏死性凋亡。
在IFN-β处理组中，碱性磷酸酶（ALP）表达降低以浓度依赖的方式抑制了基质形成。尽管IFN-β处理组和对照组之间的增殖能力没有变化，但实验组的细胞数量显著减少。TUNEL阳性细胞数量没有显著差异；然而，坏死性凋亡标志物白细胞介素-6（IL-6）和pMLKL的蛋白水平在基质形成中显著增加。在给予JAK-STAT抑制剂鲁索替尼的组中，细胞数量和ALP表达水平得到改善。此外，鲁索替尼显著抑制了IL-6和pMLKL水平。
鲁索替尼通过JAK-STAT途径干扰IFN-β介导的坏死性凋亡和成骨细胞分化。
