TU-100 (Daikenchuto) and ginger ameliorate anti-CD3 antibody induced T cell-mediated murine enteritis: microbe-independent effects involving Akt and NF-κB suppression.

Nobuhiro Ueno,Atsushi Kaneko,Chong-Zhi Wang,Yutaka Kohgo,Chun-Su Yuan,Masahiro Yamamoto,Marc Bissonnette,Toru Kono,Mark W Musch,Mikihiro Fujiya,Eugene B Chang,Takumu Hasebe,David L Boone

doi:10.1371/journal.pone.0097456

Abstract

The Japanese traditional medicine daikenchuto (TU-100) has anti-inflammatory activities, but the mechanisms remain incompletely understood. TU-100 includes ginger, ginseng, and Japanese pepper, each component possessing bioactive properties. The effects of TU-100 and individual components were investigated in a model of intestinal T lymphocyte activation using anti-CD3 antibody. To determine contribution of intestinal bacteria, specific pathogen free (SPF) and germ free (GF) mice were used. TU-100 or its components were delivered by diet or by gavage. Anti-CD3 antibody increased jejunal accumulation of fluid, increased TNFα, and induced intestinal epithelial apoptosis in both SPF and GF mice, which was blocked by either TU-100 or ginger, but not by ginseng or Japanese pepper. TU-100 and ginger also blocked anti-CD3-stimulated Akt and NF-κB activation. A co-culture system of colonic Caco2BBE and Jurkat-1 cells was used to examine T-lymphocyte/epithelial cells interactions. Jurkat-1 cells were stimulated with anti-CD3 to produce TNFα that activates epithelial cell NF-κB. TU-100 and ginger blocked anti-CD3 antibody activation of Akt in Jurkat cells, decreasing their TNFα production. Additionally, TU-100 and ginger alone blocked direct TNFα stimulation of Caco2BBE cells and decreased activation of caspase-3 and polyADP ribose. The present studies demonstrate a new anti-inflammatory action of TU-100 that is microbe-independent and due to its ginger component.

Highlights

The Japanese traditional medicine (Kampo) daikenchuto (TU100) has been established to have anti-inflammatory, prokinetic, and blood flow effects in the gastrointestinal tract in both animal models as well as humans [1,2,3,4,5,6,7,8,9,10,11,12,13,14]
Mice were gavaged with TU-100 or the individual components of TU-100: ginger, ginseng, or Japanese pepper, to determine the effects of each component
For specific pathogen-free mice (SPF contain normal intestinal microbiota) fed TU-100 diet (3 days), anti-CD3 antibody induced fluid enteropooling and distention of jejunal segments were significantly decreased (Figure 1A). To determine whether these changes were gut microbe-dependent, germ free (GF) C57Bl6 mice were fed sterile unsupplemented diet or diet supplemented with 1.5% TU-100 or gavaged with TU-100 or components that were autoclaved prior to gavage

Summary

Introduction

The Japanese traditional medicine (Kampo) daikenchuto (TU100) has been established to have anti-inflammatory, prokinetic, and blood flow effects in the gastrointestinal tract in both animal models as well as humans [1,2,3,4,5,6,7,8,9,10,11,12,13,14]. Japanese pepper contains hydroxy-sanshools (alpha and beta) that alter intestinal blood flow, motility, and barrier function by inducing adrenomedullin and calcitonin gene related peptides [3,7,8]. These compounds have been shown to activate intestinal epithelial TRPA1 channels [11]. These and other ginseng-containing compounds modulate cell growth and act as anti-cancer agents [20,21,22,23] In addition to these effects of individual extract constituents, TU-100 has been shown to activate nicotinic acetylcholine receptors, contributing to its effects on motility [13]

Methods

Results

Conclusion