Guaiacol suppresses osteoclastogenesis by blocking interactions of RANK with TRAF6 and C-Src and inhibiting NF-κB, MAPK and AKT pathways
Guaiacol suppresses osteoclastogenesis by blocking interactions of RANK with TRAF6 and C-Src and inhibiting NF-κB, MAPK and AKT pathways
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愈创木酚通过阻断 RANK 与 TRAF6 和 C-Src 的相互作用以及抑制 NF-kappa B、MAPK 和 AKT 通路来抑制破骨细胞生成
DOI：
10.1111/jcmm.15153
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发表时间：
2020-05-01
影响因子：
5.3
通讯作者：
Su, Jiacan
中科院分区：
文献类型：
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作者：
Zhi, Xin;Fang, Chao;Su, Jiacan
当归（AS；当归）是一种传统中药，几个世纪以来一直被用于治疗骨质疏松症和骨坏死等骨骼疾病。然而，其有效成分和潜在机制尚不清楚。本研究采用二维细胞膜色谱/C18柱/飞行时间质谱（2D CMC/C18柱/TOFMS）鉴定愈创木酚为as的有效成分。愈创木酚在体外以剂量依赖的方式抑制骨髓单核细胞（BMMCs）和RAW264.7细胞的破骨细胞生成和破骨细胞功能。共免疫沉淀表明愈创木酚阻断RANK-TRAF6关联和RANK-C-Src关联。愈创木酚可抑制p65、p50、I κ B （nf - κ B通路）、ERK、JNK、c-fos、p38 （MAPK通路）和Akt （Akt通路）的磷酸化，降低Cathepsin K、CTR、MMP-9和TRAP的表达水平。愈创木酚还能抑制活化t细胞胞浆核因子1（NFATc1）的表达和rankl诱导的Ca2+振荡。在体内，它通过抑制过度的破骨细胞生成来改善卵巢切除术引起的骨质流失。综上所述，我们的研究结果表明愈创木酚通过阻断RANK与TRAF6和C-Src的相互作用，以及通过抑制nf - κ B、MAPK和AKT信号通路，抑制rankl诱导的破骨细胞生成。因此，该化合物显示出治疗破骨细胞发生相关骨病的潜力，包括绝经后骨质疏松症。
Angelica sinensis (AS; Dang Gui), a traditional Chinese herb, has for centuries been used for the treatment of bone diseases, including osteoporosis and osteonecrosis. However, the effective ingredient and underlying mechanisms remain elusive. Here, we identified guaiacol as the active component of AS by two-dimensional cell membrane chromatography/C18 column/time-of-flight mass spectrometry (2D CMC/C18 column/TOFMS). Guaiacol suppressed osteoclastogenesis and osteoclast function in bone marrow monocytes (BMMCs) and RAW264.7 cells in vitro in a dose-dependent manner. Co-immunoprecipitation indicated that guaiacol blocked RANK-TRAF6 association and RANK-C-Src association. Moreover, guaiacol prevented phosphorylation of p65, p50, I kappa B (NF-kappa B pathway), ERK, JNK, c-fos, p38 (MAPK pathway) and Akt (AKT pathway), and reduced the expression levels of Cathepsin K, CTR, MMP-9 and TRAP. Guaiacol also suppressed the expression of nuclear factor of activated T-cells cytoplasmic 1(NFATc1) and the RANKL-induced Ca2+ oscillation. In vivo, it ameliorated ovariectomy-induced bone loss by suppressing excessive osteoclastogenesis. Taken together, our findings suggest that guaiacol inhibits RANKL-induced osteoclastogenesis by blocking the interactions of RANK with TRAF6 and C-Src, and by suppressing the NF-kappa B, MAPK and AKT signalling pathways. Therefore, this compound shows therapeutic potential for osteoclastogenesis-related bone diseases, including postmenopausal osteoporosis.