Connexin43 Controls the Myofibroblastic Differentiation of Bronchial Fibroblasts from Patients with Asthma

摘要

Pathologic accumulation of myofibroblasts in asthmatic bronchi is regulated by extrinsic stimuli and by the intrinsic susceptibility of bronchial fibroblasts to transforming growth factor-beta (TGF-beta). The specific function of gap junctions and connexins in this process has remained unknown. Here, we investigated the role of connexin43 (Cx43) in TGF-beta-induced myofibroblastic differentiation of fibroblasts derived from bronchoscopic biopsy specimens of patients with asthma and donors without asthma. Asthmatic fibroblasts expressed considerably higher levels of Cx43 and were more susceptible to TGF-beta(1)-induced myofibroblastic differentiation than were their nonasthmatic counterparts. TGF-beta(1) efficiently up-regulated Cx43 levels and activated the canonical Smad pathway in asthmatic cells. Ectopic Cx43 expression in nonasthmatic (Cx43(low)) fibroblasts increased their predilection to TGF-beta(1)-induced Smad2 activation and fibroblast-myofibroblast transition. Transient Cx43 silencing in asthmatic (Cx43(high)) fibroblasts by Cx43 small interfering RNA attenuated the TGF-beta(1)-triggered Smad2 activation and myofibroblast formation. Direct interactions of Smad2 and Cx43 with beta-tubulin were demonstrated by co-immunoprecipitation assay, whereas the sensitivity of these interactions to TGF-beta(1) signaling was confirmed by Forster Resonance Energy Transfer analyses. Furthermore, inhibition of the TGF-beta(1)/Smad pathway attenuated TGF-beta(1)-triggered Cx43 up-regulation and myofibroblast differentiation of asthmatic fibroblasts. Chemical inhibition of gap junctional intercellular communication with 18 alpha-glycyrrhetinic acid did not affect the initiation of fibroblast-myofibroblast transition in asthmatic fibroblasts but interfered with the maintenance of their myofibroblastic phenotype. Collectively, our data identified Cx43 as a new player in the feedback mechanism regulating TGF-beta(1)/Smad-dependent differentiation of bronchial fibroblasts. Thus, our observations point to Cx43 as a novel profibrotic factor in asthma progression.