Original Article
Copyright ©2013 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Exp Med. Nov 20, 2013; 3(4): 100-107
Published online Nov 20, 2013. doi: 10.5493/wjem.v3.i4.100
Epigallocatechin-3-gallate suppresses transforming growth factor-beta signaling by interacting with the transforming growth factor-beta type II receptor
Masaki Tabuchi, Sumio Hayakawa, Eiko Honda, Kana Ooshima, Tatsuki Itoh, Koji Yoshida, Ah-Mee Park, Hideaki Higashino, Mamoru Isemura, Hiroshi Munakata
Masaki Tabuchi, Koji Yoshida, Ah-Mee Park, Hiroshi Munakata, Department of Biochemistry, Kinki University Faculty of Medicine, Osaka-Sayama 589-8511, Japan
Sumio Hayakawa, Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan
Eiko Honda, Life Science Research Institute, Kinki University, Osaka-Sayama 589-8511, Japan
Kana Ooshima, Hideaki Higashino, Department of Pharmacology, Kinki University Faculty of Medicine, Osaka-Sayama 589-8511, Japan
Tatsuki Itoh, Department of Pathology, Kinki University Faculty of Medicine, Osaka-Sayama 589-8511, Japan
Mamoru Isemura, Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka 422-8526, Japan
Author contributions: Tabuchi M and Hayakawa S contributed equally to this work; Tabuchi M and Hayakawa S designed studies; Tabuchi M, Hayakawa S, Honda E, Ooshima K, Itoh T, Yoshida K, Park AM, Higashino H and Isemura M performed the research; Tabuchi M, Hayakawa S and Munakata H analyzed the data; Munakata H wrote the paper.
Correspondence to: Hiroshi Munakata, Professor, Department of Biochemistry, Kinki University Faculty of Medicine, 377-2 Ohnohigashi, Osaka-Sayama 589-8511, Japan. munakata@med.kindai.ac.jp
Telephone: +81-72-3660221 Fax: +81-72-3660245
Received: May 14, 2013
Revised: July 12, 2013
Accepted: September 18, 2013
Published online: November 20, 2013
Processing time: 190 Days and 7.2 Hours
Abstract

AIM: To investigate the (-)-epigallocatechin-3-gallate (EGCG) binding to transforming growth factor-β (TGF-β) type II receptor (TGFRII).

METHODS: The expression of α-smooth muscle actin (α-SMA) was used as a marker for fibrotic change in human lung fibroblast MRC-5 cells. The α-SMA expression level was determined by western blotting and immunohistological analysis. We examined whether the anti-fibrotic effects of EGCG on MRC-5 cells was dependent on antioxidant mechanism by using edaravone and N-acetylcysteine (NAC). The suppression effects of EGCG on Smad2/3 activation were studied by confocal fluorescence microscopy. The binding of EGCG to recombinant TGFRII protein was analyzed by immunoprecipitation and affinity chromatography.

RESULTS: When MRC-5 cells were treated with TGF-β, EGCG decreased the expression of α-SMA in a dose dependent manner, whereas catechin did not influence the α-SMA expression in the cells. Except for EGCG, antioxidant compounds (e.g., edaravone and NAC) had no effects on the TGF-β-induced α-SMA expression. Nuclear localization of phosphorylated Smad2/3 was observed after TGF-β treatment; however, EGCG treatment attenuated the nuclear transportation of Smad2/3 in the presence or absence of TGF-β. After a TGFRII expression vector was introduced into COS-7 cells, cell lysates were untreated or treated with EGCG or catechin. The immunoprecipitation experiments using the lysates showed that EGCG dose-dependently bound to TGFRIIand that catechin did not at all. Affinity chromatography study indicated that EGCG would bind to TGFRII.

CONCLUSION: Our results demonstrate that EGCG interacts with TGFRII and inhibits the expression of α-SMA via the TGF-β-Smad2/3 pathway in human lung fibroblast MRC-5 cells.

Keywords: Epigallocatechin-3-gallate; Transforming growth factor-β; Myofibroblast; α-smooth muscle actin; Fibrosis

Core tip: (-)-Epigallocatechin-3-gallate (EGCG) binds to transforming growth factor-β (TGF-β) type II receptor (TGFRII) and inhibits TGF-β action by interfering with the interaction between TGF-β and TGFRII. Because TGF-β is considered to be the strongest inducer of tissue fibrosis, the obtained data from this investigation suggest that EGCG may be a new therapeutic agent for organ fibrosis.