yam63(1)_79.pdf 1.9 MB
Sakaguchi, Takuki Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medical Sciences, Tottori University / Division of Medicine and Clinical Science, Department of Multidisciplinary Internal Medicine, School of Medicine, Tottori University Faculty of Medicine
Kono, Yohei Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medical Sciences, Tottori University
Itaba, Noriko Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medical Sciences, Tottori University Researchers DB KAKEN
Isomoto, Hajime Division of Medicine and Clinical Science, Department of Multidisciplinary Internal Medicine, School of Medicine, Tottori University Faculty of Medicine Researchers DB KAKEN
hepatic stellate cells
Background: Liver fibrosis progresses to decompensated liver cirrhosis, for which medical needs remain unmet. We recently developed IC-2, a small-molecule compound that suppresses Wnt/β-catenin signaling, and found that IC-2 also suppresses liver fibrosis. In this study, we performed three-step screening of newly synthesized IC-2 derivatives to identify other small-molecule compounds that suppress liver fibrosis.
Methods: The screening system consisted of three steps: a cell viability assay, a transcription factor 4 (TCF4) reporter assay, and induction of α-smooth muscle actin (α-SMA) and collagen 1α1 (Col1A1) expression in response to each compound. Screening using human LX-2 hepatic stellate cells (HSCs) was performed to target HSCs, which are the driver cells of liver fibrosis.
Results: In the first step, since 9b and 9b-CONH2 at 100 μM did not have any effects on cell viability, they were omitted in the next screening. Additionally, the conditions that led to > 40% inhibition of the controls were also excluded in subsequent screening. The second step was performed under 31 conditions for 19 small-molecule compounds. Sixteen small-molecule compounds caused significant reduction of TCF4 activity relative to that of 0.1% DMSO. Of the 16 compounds, the 10 showing the greatest suppression of TCF4 activity were selected for the third step. Expressions of mRNA for α-SMA and Col1A1 were significantly reduced by seven and three small-molecule compounds, respectively. The greatest reductions in the α-SMA and Col1A1 mRNA expressions were observed in the cells treated with IC-2-F. Protein expressions of α-SMA and Col1A1 caused by IC-2-F were also comparable to those caused by IC-2.
Conclusion: IC-2-F was identified as a novel deactivating small-molecule compound for HSCs in vitro. These data suggest that IC-2-F is a promising medicine for liver fibrosis.
Tottori University Medical Press
Yonago Acta Medica
|Current Journal Title||
Yonago Acta Medica
注があるものを除き、この著作物は日本国著作権法により保護されています。 / This work is protected under Japanese Copyright Law unless otherwise noted.
Takuki Sakaguchi, Yohei Kono, Noriko Itaba, Minoru Morimoto, Hajime Isomoto, Goshi Shiota, Identification of a Novel Deactivating Small-Molecule Compound for Fibrogenic Hepatic Stellate Cells, Yonago Acta Medica, 2020, Volume 63, Issue 1, Pages 79-87, Released February 20, 2020
Faculty of Medicine/Graduate School of Medical Sciences/University Hospital