Induction of HO-1 by Mevastatin Mediated via a Nox/ROS-Dependent c-Src/PDGFRα/PI3K/Akt/Nrf2/ARE Cascade Suppresses TNF-α-Induced Lung Inflammation
Induction of HO-1 by Mevastatin Mediated via a Nox/ROS-Dependent c-Src/PDGFRα/PI3K/Akt/Nrf2/ARE Cascade Suppresses TNF-α-Induced Lung Inflammation
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DOI：
10.3390/jcm9010226
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发表时间：
2020-01-01
影响因子：
3.9
通讯作者：
Yang, Chuen-Mao
中科院分区：
文献类型：
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作者：
Lin, Chih-Chung;Lin, Wei-Ning;Yang, Chuen-Mao
背景：美伐他汀(MVS)是一种3-羟基-3-甲基戊二酰辅酶，是一种还原酶(HMG-CoA)抑制剂，可能通过上调血红素加氧酶-1(HO-1)而具有抗炎作用。然而，MVS诱导HO-1在人肺泡上皮细胞(HPAEpiCs)中表达的机制仍很不清楚。方法：采用实时定量聚合酶链式反应、Western blotting和启动子报告分析等方法检测HO-1和ICAM-1的表达。用药物抑制剂或特异性小干扰核糖核酸(SIRNA)S研究NRF2与HO-1启动子的抗氧化反应元件(ARE)结合部位之间的相互作用。结果：MVS上调HO-1可减弱肿瘤坏死因子α刺激的ICAM-1的表达，这种表达与THP-1与HPAEpiCs的黏附有关。锡原卟啉(SnPP)IX或HO-1 siRNA可逆转HO-1的上述抑制作用。MVS诱导的HO-1表达是通过NADPH氧化酶(NOx)产生的活性氧(ROS)介导的。NOX2/ROS的激活进一步刺激P47(Phox)、原癌基因酪氨酸蛋白激酶(c-Src)、血小板衍生生长因子受体(PDFGR)α、蛋白激酶B(Akt)和Nrf2的磷酸化，而这些被siRNAs抑制。药物抑制剂二苯碘、载脂蛋白、N-乙酰-L半胱氨酸、PP1、AG1296或LY294002可降低MVS激活的Nrf2核转位与HO-1启动子上ARE的结合。结论：MVS诱导的HO-1至少部分是通过依赖于p47(Phox)/NOX2/ROS激活c-Src/PDGFRα/PI3K/Akt调节的Nrf2/ARE轴，抑制肿瘤坏死因子-α介导的炎症反应。
Background: Mevastatin (MVS), a 3-hydroxy-3-methylglutaryl coenzyme, a reductase (HMG-CoA) inhibitor, has anti-inflammatory effects potentially via up-regulation of heme oxygenase-1 (HO-1). However, the mechanisms underlying MVS-induced HO-1 expression remain largely unknown in human pulmonary alveolar epithelial cells (HPAEpiCs). Methods: HO-1 and intercellular adhesion molecule (ICAM)-1 expression were determined using real-time PCR, Western blotting, and promoter reporter analyses. The signaling components were investigated using pharmacological inhibitors or specific small interfering RNA (siRNA)s. Interaction between Nrf2 and the antioxidant response element (ARE) binding site for the HO-1 promoter was determined by chromatin immunoprecipitation (ChIP) assay. Results: Upregulation of HO-1 by MVS attenuated the tumor necrosis factor (TNF)-alpha-stimulated ICAM-1 expression associated with THP-1 adhesion to HPAEpiCs. These inhibitory effects of HO-1 were reversed by tin protoporphyrin (SnPP)IX or by transfection with HO-1 siRNA. MVS-induced HO-1 expression was mediated via NADPH oxidase (Nox)-derived reactive oxygen species (ROS) generation. Activation of Nox2/ROS further stimulated the phosphorylation of p47(phox), proto-oncogene tyrosine-protein kinase (c-Src), platelet-derived growth factor receptor (PDFGR)alpha, protein kinase B (Akt), and Nrf2, which were inhibited by siRNAs. Pretreatment with pharmacological inhibitors, including diphenyleneiodonium (DPI), apocynin (APO), N-acetyl-L-cysteine (NAC), PP1, AG1296, or LY294002, reduced the MVS-activated Nrf2 nuclear-translocation binding to the ARE on the HO-1 promoter. Conclusions: MVS-induced HO-1 is, at least in part, mediated through a p47(phox)/Nox2/ROS-dependent activation of c-Src/PDGFR alpha/PI3K/Akt-regulated Nrf2/ARE axis and suppresses the TNF-alpha-mediated inflammatory responses in HPAEpiCs.