Suppressive activity of tiotropium bromide on matrix metalloproteinase production from lung fibroblasts in vitro

Abstract

Kazuhito Asano1, Yusuke Shikama2, Yasuhiro Shibuya2, Hiroaki Nakajima2, Ken-ichi Kanai3, Naohiro Yamada3, Harumi Suzaki31Division of Physiology, School of Nursing and Rehabilitation Sciences; 2Respiratory Disease Center, Showa University Northern Yokohama Hospital, Yokohama, Japan; 3Department of Otolaryngology, School of Medicine, Showa University, Tokyo, JapanBackground: Chronic obstructive pulmonary disease (COPD) is characterized by airway remodeling with an accumulation of inflammatory cells. There is also increasing evidence that metalloproteinases (MMPs) may contribute to the pathogenesis of COPD, but the influence of agents that used for the treatment of COPD is not well understood.Objective: We evaluated whether tiotropium bromide hydrate (TBH), a M3 muscarinic receptor antagonist, could inhibit MMP production from lung fibroblasts (LFs) in response to tumor necrosis factor (TNF)-α stimulation.Methods: LFs were established from normal lung tissues taken from patients with lung tumors. LFs (5 × 105 cells/ml) were stimulated with TNF-α in the presence of various concentrations of TBH. After 24 h, culture supernatants were obtained and assayed for the levels of MMPs and tissue inhibitor of metalloproteinases (TIMPs) by ELISA. The influence of TBH on mRNA expression of MMPs and TIMPs in 4 h-cultured cells was also examined by real-time RT-PCR. Furthermore, nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) in LFs treated with TBH for 4 h was examined by ELISA.Results: TBH at more than 15 pg/ml inhibited the production of MMP-2 from LFs after TNF-α stimulation, whereas TIMP-1 and TIMP-2 production was scarcely affected by TBH through the suppression of both mRNA expression and transcription factor, NF-κB, activation in LFs induced by TNF-α stimulation.Conclusion: These results suggest that the attenuating effect of TBH on MMP-2 production from LFs induced by inflammatory stimulation may be additional beneficial therapeutic effects not directly relating to its bronchodilatory effects.Keywords: tiotropium, lung fibroblasts, matrix metalloproteinases, suppression, in vitro