Sphingosine Kinase 1/S1P Signaling Contributes to Pulmonary Fibrosis by Activating Hippo/YAP Pathway and Mitochondrial Reactive Oxygen Species in Lung Fibroblasts
Sphingosine Kinase 1/S1P Signaling Contributes to Pulmonary Fibrosis by Activating Hippo/YAP Pathway and Mitochondrial Reactive Oxygen Species in Lung Fibroblasts
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DOI：
10.3390/ijms21062064
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发表时间：
2020-03-01
影响因子：
5.6
通讯作者：
Natarajan, Viswanathan
中科院分区：
文献类型：
--
作者：
Huang, Long Shuang;Sudhadevi, Tara;Natarajan, Viswanathan
鞘氨醇激酶1（SPHK 1）/鞘氨醇-1-磷酸（S1 P）信号传导轴正在成为小鼠特发性肺纤维化（IPF）和博莱霉素（BLM）诱导的肺纤维化发展的关键参与者。最近的证据表明Hippo/Yes相关蛋白（雅普）1通路参与了肺部疾病，包括IPF，但其与SPHK 1/S1 P信号通路的合理联系尚不清楚。在此，我们证明了BLM攻击小鼠肺成纤维细胞中YAP 1与成纤维细胞标志物FSP 1的共定位增加，并且小鼠肺成纤维细胞（MLF）中Sphk 1的遗传缺失减少了YAP 1在纤维化灶中的定位。PF 543对小鼠SPHK 1活性的抑制减弱了肺成纤维细胞中YAP 1与FSP 1的共定位。在体外，TGF-β刺激YAP 1易位到原代MLF的细胞核，Sphk 1的缺失或PF 543的抑制减弱了TGF-β介导的YAP 1核定位。此外，PF 543抑制SPHK 1或维替泊芬抑制YAP 1可降低人肺成纤维细胞（HLF）中TGF-β或BLM诱导的线粒体活性氧（mtROS）以及纤连蛋白（FN）和α-平滑肌肌动蛋白（α-SMA）的表达。此外，用MitoTEMPO清除mtROS减弱了TGF-β诱导的FN和α-SMA表达。向HLF中添加S1 P抗体可减少TGF-β或S1 P介导的YAP 1激活、mtROS以及FN和alpha-SMA的表达。这些结果表明SPHK 1/S1 P信号在TGF-β诱导的YAP 1活化和mtROS产生中的作用，导致成纤维细胞活化，这是肺纤维化的关键驱动因素。
The sphingosine kinase 1 (SPHK1)/sphingosine-1-phosphate (S1P) signaling axis is emerging as a key player in the development of idiopathic pulmonary fibrosis (IPF) and bleomycin (BLM)-induced lung fibrosis in mice. Recent evidence implicates the involvement of the Hippo/Yes-associated protein (YAP) 1 pathway in lung diseases, including IPF, but its plausible link to the SPHK1/S1P signaling pathway is unclear. Herein, we demonstrate the increased co-localization of YAP1 with the fibroblast marker FSP1 in the lung fibroblasts of BLM-challenged mice, and the genetic deletion of Sphk1 in mouse lung fibroblasts (MLFs) reduced YAP1 localization in fibrotic foci. The PF543 inhibition of SPHK1 activity in mice attenuated YAP1 co-localization with FSP1 in lung fibroblasts. In vitro, TGF-beta stimulated YAP1 translocation to the nucleus in primary MLFs, and the deletion of Sphk1 or inhibition with PF543 attenuated TGF-beta-mediated YAP1 nuclear localization. Moreover, the PF543 inhibition of SPHK1, or the verteporfin inhibition of YAP1, decreased the TGF-beta- or BLM-induced mitochondrial reactive oxygen species (mtROS) in human lung fibroblasts (HLFs) and the expression of fibronectin (FN) and alpha-smooth muscle actin (alpha-SMA). Furthermore, scavenging mtROS with MitoTEMPO attenuated the TGF-beta-induced expression of FN and alpha-SMA. The addition of the S1P antibody to HLFs reduced TGF-beta- or S1P-mediated YAP1 activation, mtROS, and the expression of FN and alpha-SMA. These results suggest a role for SPHK1/S1P signaling in TGF-beta-induced YAP1 activation and mtROS generation, resulting in fibroblast activation, a critical driver of pulmonary fibrosis.