Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan.
Department of Medical Materials for Musculoskeletal Reconstruction, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan.
Inflamm Res. 2017 Nov;66(11):999-1009. doi: 10.1007/s00011-017-1083-x. Epub 2017 Jul 27.
To investigate whether janus kinase (JAK) inhibitor exhibits a chondro-protective effect against mechanical stress-induced expression of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) and matrix metalloproteinase (MMPs) in human chondrocytes.
Normal human articular chondrocytes were seeded onto stretch chambers and incubated with or without tofacitinib (1000 nM) for 12 h before mechanical stimulation or cytokine stimulation. Uni-axial cyclic tensile strain (CTS) (0.5 Hz, 10% elongation, 30 min) was applied and the gene expression levels of type II collagen α1 chain (COL2A1), aggrecan (ACAN), ADAMTS4, ADAMTS5, MMP13, and runt-related transcription factor 2 (RUNX-2) were examined by real-time polymerase chain reaction. Nuclear translocation of RUNX-2 and nuclear factor-κB (NF-κB) was examined by immunocytochemistry, and phosphorylation of mitogen-activated protein kinase (MAPK) and signaling transducer and activator of transcription (STAT) 3 was examined by western blotting. The concentration of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α in the supernatant was examined by enzyme-linked immunosorbent assay.
COL2A1 and ACAN gene expression levels were decreased by CTS, but these catabolic effects were canceled by tofacitinib. Tofacitinib significantly down-regulated CTS-induced expression of ADAMTS4, ADAMTS5, MMP13, and RUNX2, and the release of IL-6 in supernatant by chondrocytes. Tofacitinib also reduced CTS-induced nuclear translocation of RUNX-2 and NF-κB, and phosphorylation of MAPK and STAT3.
Tofacitinib suppressed mechanical stress-induced expression of ADAMTS4, ADAMTS5, and MMP13 by human chondrocytes through inhibition of the JAK/STAT and MAPK cascades.
研究 Janus 激酶（JAK）抑制剂是否对机械应力诱导的人类软骨细胞中解整合素金属蛋白酶与凝血酶 3 型（ADAMTS）和基质金属蛋白酶（MMPs）的表达具有软骨保护作用。
将正常的人关节软骨细胞接种到拉伸室中，在机械刺激或细胞因子刺激前用或不用托法替尼（1000 nM）孵育 12 小时。施加单轴循环拉伸应变（CTS）（0.5 Hz，10%伸长，30 分钟），通过实时聚合酶链反应检测 II 型胶原α1 链（COL2A1）、聚集蛋白（ACAN）、ADAMTS4、ADAMTS5、MMP13 和 runt 相关转录因子 2（RUNX-2）的基因表达水平。通过免疫细胞化学检测 RUNX-2 和核因子-κB（NF-κB）的核转位，通过蛋白质印迹法检测丝裂原激活蛋白激酶（MAPK）和信号转导和转录激活因子（STAT）3 的磷酸化。通过酶联免疫吸附试验检测上清液中白细胞介素（IL）-1β、IL-6 和肿瘤坏死因子-α的浓度。
CTS 降低了 COL2A1 和 ACAN 的基因表达水平，但托法替尼可消除这些分解代谢作用。托法替尼显著下调 CTS 诱导的 ADAMTS4、ADAMTS5、MMP13 和 RUNX2 表达，以及上清液中 IL-6 的释放。托法替尼还降低了 CTS 诱导的 RUNX-2 和 NF-κB 的核转位，以及 MAPK 和 STAT3 的磷酸化。
托法替尼通过抑制 JAK/STAT 和 MAPK 级联反应，抑制人软骨细胞机械应力诱导的 ADAMTS4、ADAMTS5 和 MMP13 的表达。
