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Am. J. Respir. Cell Mol. Biol., Volume 21, Number 3, September, 1999 428-435

Protective Effects of Heme Oxygenase-1 against Oxidant-Induced Injury in the Cultured Human Tracheal Epithelium

Norihiro Yamada, Mutsuo Yamaya, Shoji Okinaga, Ria Lie, Tomoko Suzuki, Katsutoshi Nakayama, Atsushi Takeda, Tokio Yamaguchi, Yasuto Itoyama, Kiyohisa Sekizawa, and Hidetada Sasaki

Department of Geriatric Medicine and Neurology, Tohoku University School of Medicine, Sendai, Japan

To examine whether increases in heme oxygenase (HO)-1 activity have protective effects on the oxidant-induced injury of airway epithelial cells, human tracheal epithelial cells were cultured on a porous filter membrane, and electrical conductance (G) and mannitol flux across epithelial membrane were measured with Ussing's chamber methods and D-[3H]mannitol, respectively. Hydrogen peroxide (H2O2; 1 mM) increased G with time from the baseline value of 6.0 ± 0.6 to 17.8 ± 0.9 mS/cm2 at 6 h after administration (P < 0.001). Likewise, H2O2 significantly increased mannitol flux through the cultured epithelium (P < 0.01). Pretreatment of cultured epithelial cells with hemin (10 µM; 8 h) or interleukin (IL)-1beta (10 ng/ml; 16 h) completely inhibited increases in G and mannitol flux induced by H2O2. Tin protoporphyrin IX (50 µm) and zinc protoporphyrin IX (10 µM), inhibitors of HO-1, reduced hemin-induced and IL-1beta -induced inhibitory effects. Hemin treatment increased HO-1 messenger RNA expression, HO-1 protein production, and HO activity and bilirubin content as well as ferritin content in the cultured epithelial cells. Pretreatment with hemin and desferoxamine, which, like ferritin, can bind iron, inhibited H2O2-induced increases in G and mannitol permeability. Although exogenous bilirubin mimicked hemin-induced inhibitory effects, exogenous apoferritin failed to inhibit H2O2-induced effects on G and mannitol permeability. These findings suggest that HO-1 induction provides protection against H2O2-induced injury of the cultured human airway epithelial cells in part via the HO-bilirubin pathway.




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