caa a22 4500 606176144 CHVBK 20210128100746.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00018-014-1798-6 doi (NATIONALLICENCE)springer-10.1007/s00018-014-1798-6 EW-7197 inhibits hepatic, renal, and pulmonary fibrosis by blocking TGF-β/Smad and ROS signaling [Elektronische Daten] [Sang-A Park, Min-Jin Kim, So-Yeon Park, Jung-Shin Kim, Seon-Joo Lee, Hyun Woo, Dae-Kee Kim, Jeong-Seok Nam, Yhun Sheen] Fibrosis is an inherent response to chronic damage upon immense apoptosis or necrosis. Transforming growth factor-beta1 (TGF-β1) signaling plays a key role in the fibrotic response to chronic liver injury. To develop anti-fibrotic therapeutics, we synthesized a novel small-molecule inhibitor of the TGF-β type I receptor kinase (ALK5), EW-7197, and evaluated its therapeutic potential in carbon tetrachloride (CCl4) mouse, bile duct ligation (BDL) rat, bleomycin (BLM) mouse, and unilateral ureteral obstruction (UUO) mouse models. Western blot, immunofluorescence, siRNA, and ChIP analysis were carried out to characterize EW-7197 as a TGF-β/Smad signaling inhibitor in LX-2, Hepa1c1c7, NRK52E, and MRC5 cells. In vivo anti-fibrotic activities of EW-7197 were examined by microarray, immunohistochemistry, western blotting, and a survival study in the animal models. EW-7197 decreased the expression of collagen, α-smooth muscle actin (α-SMA), fibronectin, 4-hydroxy-2, 3-nonenal, and integrins in the livers of CCl4 mice and BDL rats, in the lungs of BLM mice, and in the kidneys of UUO mice. Furthermore, EW-7197 extended the lifespan of CCl4 mice, BDL rats, and BLM mice. EW-7197 blocked the TGF-β1-stimulated production of reactive oxygen species (ROS), collagen, and α-SMA in LX-2 cells and hepatic stellate cells (HSCs) isolated from mice. Moreover, EW-7197 attenuated TGF-β- and ROS-induced HSCs activation to myofibroblasts as well as extracellular matrix accumulation. The mechanism of EW-7197 appeared to be blockade of both TGF-β1/Smad2/3 and ROS signaling to exert an anti-fibrotic activity. This study shows that EW-7197 has a strong potential as an anti-fibrosis therapeutic agent via inhibition of TGF-β-/Smad2/3 and ROS signaling. Springer Basel, 2014 ALK5 inhibitor nationallicence CCl4 hepatic fibrosis nationallicence BDL hepatic fibrosis nationallicence UUO renal fibrosis nationallicence BLM pulmonary fibrosis nationallicence 4-HNE : 4-Hydroxy-2,3-nonenal nationallicence α-SMA : Alpha-smooth muscle actin nationallicence ALK5 : Activin receptor-like kinase 5 nationallicence BDL : Bile duct ligation nationallicence BLM : Bleomycin nationallicence CCl4 : Carbon tetrachloride nationallicence ECM : Extracellular matrix nationallicence EW-7197 : N-[[4-([1,2,4]Triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl]methyl]-2-fluoroaniline nationallicence GOx : Glucose oxidase nationallicence HSCs : Hepatic stellate cells nationallicence IHC : Immunohistochemistry nationallicence NOX : Nicotinamide adenine dinucleotide phosphate oxidase nationallicence Prdx : Peroxiredoxin nationallicence qRT-PCR : Quantitative real-time reverse-transcriptase polymerase chain reaction nationallicence ROS : Reactive oxygen species nationallicence TGF-β : Transforming growth factor-beta nationallicence UUO : Unilateral ureteral obstruction nationallicence Park Sang-A College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Kim Min-Jin College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Park So-Yeon College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Kim Jung-Shin College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Lee Seon-Joo College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Woo Hyun College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Kim Dae-Kee College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Nam Jeong-Seok Laboratory of Tumor Suppressor, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, 406-840, Incheon, South Korea aut Sheen Yhun College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut Cellular and Molecular Life Sciences Springer Basel 72/10(2015-05-01), 2023-2039 1420-682X 72:10<2023 2015 72 18 https://doi.org/10.1007/s00018-014-1798-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-014-1798-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Park Sang-A College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Min-Jin College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Park So-Yeon College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Jung-Shin College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Lee Seon-Joo College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Woo Hyun College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Dae-Kee College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 700 1- Nam Jeong-Seok Laboratory of Tumor Suppressor, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, 406-840, Incheon, South Korea aut NATIONALLICENCE 700 1- Sheen Yhun College of Pharmacy, Ewha Womans University, Seodaemun-gu, 120-750, Seoul, South Korea aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/10(2015-05-01), 2023-2039 1420-682X 72:10<2023 2015 72 18