Department of Orthopedics, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Oncology, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
J Cell Physiol. 2019 May;234(5):6831-6841. doi: 10.1002/jcp.27432. Epub 2018 Nov 1.
The aim of this study is to evaluate the effect of transient receptor potential vanilloid 4 (TRPV4) on osteoclast differentiation and osteoporosis, and to investigate the underlying mechanism. The results showed that TRPV4 expression and intracellular Ca concentration were significantly upregulated in macrophage colony-stimulating factor (M-CSF)-stimulated and receptor activator of nuclear factor κΒ ligand (RANKL)-stimulated RAW264.7 cells. Furthermore, TRPV4 overexpression further increased the M-CSF- and RANKL-induced number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts and expression of osteoclastogenesis-related genes (TRAP, c-Fos, and nuclear factor of activated T cells [NFATc1]), activated the Ca -calcineurin-NFATc1 signaling and increased autophagy-related proteins (light chain [LC] 3II and Beclin-1) during osteoclast differentiation. In contrast, TRPV4 knockdown exerted the opposite effects. Mechanically, inhibition of Ca -calcineurin-NFATc1 signaling by FK506 or 11R-VIVIT abrogated the TRPV4 overexpression-induced osteoclast differentiation and autophagy induction. Moreover, suppression of autophagy by 3-methyladenine attenuated the TRPV4-induced osteoclast differentiation. In addition, short hairpin RNA TRPV4-lentivirus administration significantly diminished the increased levels of several osteoclastogenesis-related genes (RANKL, TRAP, and tumor necrosis factor-α), alleviated the disturbed microarchitecture of lumbar vertebrae, restored the decreased bone mineral density, ratio of bone volume to total tissue volume, trabecular thickness, and trabecular number, and diminished the increased trabecular separation, in ovariectomy (OVX)-induced osteoporosis mice. Consistent with the in vitro data, TRPV4 knockdown significantly decreased the induced number of TRAP-positive osteoclasts, the increased LC3 and NFATc1 expression in the lumbar vertebrae of OVX mice. In conclusion, TRPV4 knockdown suppresses osteoclast differentiation and osteoporosis by inhibiting autophagy through Ca -calcineurin-NFATc1 pathway.
本研究旨在评估瞬时受体电位香草酸 4（TRPV4）对破骨细胞分化和骨质疏松症的影响，并探讨其潜在机制。结果表明，在巨噬细胞集落刺激因子（M-CSF）刺激和核因子κΒ 配体（RANKL）刺激的 RAW264.7 细胞中，TRPV4 表达和细胞内 Ca 浓度显著上调。此外，TRPV4 过表达进一步增加了 M-CSF 和 RANKL 诱导的抗酒石酸酸性磷酸酶（TRAP）阳性破骨细胞数量和破骨细胞分化相关基因（TRAP、c-Fos 和激活 T 细胞核因子 [NFATc1]）的表达，激活了 Ca -钙调神经磷酸酶-NFATc1 信号通路，并增加了自噬相关蛋白（LC3II 和 Beclin-1）在破骨细胞分化过程中的表达。相反，TRPV4 敲低则产生相反的效果。机制上，FK506 或 11R-VIVIT 抑制 Ca -钙调神经磷酸酶-NFATc1 信号通路可阻断 TRPV4 过表达诱导的破骨细胞分化和自噬诱导。此外，3-甲基腺嘌呤抑制自噬可减弱 TRPV4 诱导的破骨细胞分化。此外，短发夹 RNA TRPV4-慢病毒给药可显著降低几种破骨细胞分化相关基因（RANKL、TRAP 和肿瘤坏死因子-α）的上调水平，缓解腰椎微结构紊乱，恢复骨密度、骨体积与组织总体积比、骨小梁厚度和骨小梁数量的降低，并减少骨小梁分离的增加，在卵巢切除（OVX）诱导的骨质疏松症小鼠中。与体外数据一致，TRPV4 敲低显著减少了诱导的 TRAP 阳性破骨细胞数量，以及 OVX 小鼠腰椎中 LC3 和 NFATc1 表达的增加。总之，TRPV4 敲低通过 Ca -钙调神经磷酸酶-NFATc1 通路抑制自噬来抑制破骨细胞分化和骨质疏松症。
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