Hydrogen sulphide promotes osteoclastogenesis by inhibiting autophagy through the PI3K/AKT/mTOR pathway
Hydrogen sulphide promotes osteoclastogenesis by inhibiting autophagy through the PI3K/AKT/mTOR pathway
复制标题
硫化氢通过 PI3K/AKT/mTOR 途径抑制自噬，促进破骨细胞生成
DOI：
10.1080/1061186x.2019.1624969
复制
发表时间：
2019-06-25
影响因子：
4.5
通讯作者：
Ye, TianWen
中科院分区：
文献类型：
--
作者：
Ma, Jun;Du, Di;Ye, TianWen
摘要硫化氢（H2S）是一种气体递质，在调节骨稳态中起重要作用。然而，H2S对破骨细胞的确切影响仍然是难以捉摸的。本研究的目的是确定H2S在调节破骨细胞中的潜在作用以及H2S影响破骨细胞生成的潜在机制。本研究应用了western blot、实时荧光定量PCR、抗酒石酸酸性磷酸酶染色、小凹形成试验、免疫荧光共聚焦显微镜、透射电镜和annexin V-异硫氰酸荧光素/碘化丙啶双染试验。结果表明，在核因子κ-β受体激活剂配体（RANKL）诱导的破骨细胞分化过程中，胱硫醚b合酶（CBS）和胱硫醚c裂解酶（CSE）的表达明显增加。此外，H2S促进RANKL诱导的破骨细胞生成并抑制成熟破骨细胞的凋亡。机制上，H2S抑制Raw 264.7细胞中的自噬。自噬激活剂（雷帕霉素）减轻了H2S对破骨细胞分化的诱导作用。进一步的研究表明，H2S通过激活PI 3 K/AKT/mTOR信号通路抑制自噬。综上所述，我们的研究结果表明，外源性H2S可以通过激活PI 3 K/AKT/mTOR通路来促进破骨细胞的发生，从而下调自噬。
Abstract Hydrogen sulphide (H2S), a gasotransmitter, plays an important role in the regulation of bone homeostasis. However, the precise effect of H2S on osteoclasts was still elusive. The goal of this study was to determine the potential role of H2S in regulation of osteoclasts and the underlying mechanisms by which H2S affected osteoclastogenesis. The present study applied western blot, quantitative real-time PCR, tartrate-resistant acid phosphatase staining, pit formation assay, immunofluorescence confocal microscopy, transmission electron microscopy and annexin V-fluorescein isothiocyanate/propidium iodide double-staining assay. The results showed that the expressions of cystathionine b-synthase (CBS) and cystathionine c-lyase (CSE) were obviously increased in osteoclast differentiation induced by the receptor activator of nuclear factor kappa-β ligand (RANKL). In addition, H2S promoted RANKL-induced osteoclastogenesis and inhibited apoptosis of mature osteoclasts. Mechanistically, H2S inhibited autophagy in Raw 264.7 cells. Autophagy activator (rapamycin) alleviated the induction of osteoclast differentiation by H2S. Further studies showed that H2S inhibited autophagy by activating the PI3K/AKT/mTOR signalling pathway. Taken together, our results implicated that exogenous H2S could promote osteoclastogenesis by activating the PI3K/AKT/mTOR pathway to downregulate autophagy.