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
Authors: Rou-Ling Cho
Hui-Ching Tseng
Chien-Chung Yang
Yi-Cheng Yeh
Chih-Chung Lin
Wei-Ning Lin
Chuen-Mao Yang
Li-Der Hsiao
Journal: Journal of clinical medicine (2020)
Discipline: CLINICAL MEDICINE
Keywords: AREs; Nrf2; ROS; heme oxygenase-1; mevastatin.
Affiliations: Department of Anesthetics, Chang Gung Memorial Hospital at Linkuo, Kwei-San, Tao-Yuan 33302, Taiwan., Graduate Institution of Biomedical and Pharmaceutical Science, College of Medicine, Fu Jen Catholic University, New Taipei City 24205, Taiwan., Department of Pharmacology, College of Medicine, China Medical University, Taichung 40402, Taiwan., Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital at Tao-Yuan, Kwei-San, Tao-Yuan 33302, Taiwan., School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan 33302, Taiwan., Department of Post-Baccalaureate Veterinary Medicine, College of Medical and Health Science, Asia University, Wufeng, Taichung 41354, Taiwan.

Abstract:  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)-α-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 p47phox, proto-oncogene tyrosine-protein kinase (c-Src), platelet-derived growth factor receptor (PDFGR)α, 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 p47phox/Nox2/ROS-dependent activation of c-Src/PDGFRα/PI3K/Akt-regulated Nrf2/ARE axis and suppresses the TNF-α-mediated inflammatory responses in HPAEpiCs. 

Retracted: No

PubPeer comments:
 FIGURE 10  Induction of apoptosis in HepG2 cells by complex 3 examined using fluorescenceu2010activated cell sorting analysis with PI staining after treatment for 24 h. (a) Control, (b) complex 3 at 5 u03bcM and (c) complex 3 at 10 u03bcM . The only discernable difference between (a) and (b) is in the percentage count for Q4. ![file] , 
 FIGURE 11  Fluorescenceu2010activated cell sorting analyses based on DNA content and stained by PI for 24 h . Three separately-treated cell cultures produced FACS histograms with identical G1 humps. ![file] Histograms should not be eroded and undercut like the Gedu00e4chtniskirche in Berlin. ![file] , 
 FIGURE 9  Agarose gel electrophoresis pattern for the cleavage of pBR322 plasmid DNA (10 u03bcM) by complexes 1u20134 at 37 u00b0C after incubation for 45 min at 25 u03bcM concentration. Lane 1: DNA control; lane 2: (100 u03bcM) CuCl2 + DNA; lane 3: (100 u03bcM) ZnCl2 + DNA; lane 4: complex 1 + DNA; lane 5: complex 2 + DNA; lane 6: complex 3 + DNA; lane 7: complex 4 + DNA . ![file] ,