title: Endothelial connexin43 mediates acid-induced increases in pulmonary microvascular permeability.
abstract: Acid aspiration, a common cause of acute lung injury, leads to alveolar edema. Increase in lung vascular permeability underlies this pathology. To define mechanisms, isolated rat lungs were perfused with autologous blood. Hydrochloric acid and rhodamine-dextran 70 kDa (RDx70) were coinstilled into an alveolus by micropuncture. RDx70 fluorescence was used to establish the spatial distribution of acid. Subsequently, FITC-dextran 20 kDa (FDx20) was infused into microvessels for 60 min followed by a 10-min HEPES-buffered saline wash. During the infusion, FITC fluorescence changes were recorded to quantify the ratio of peak to postwash fluorescence. The ratio, termed normalized fluorescence, was low for acid compared with buffer instillation both in microvessels abutting acid-treated alveoli and those located more than 700 μm away. In contrast, the normalized fluorescence was similar to buffer controls when a higher molecular weight tracer (FITC-dextran 70 kDa) was infused instead of FDx20, suggesting that normalized FDx20 fluorescence faithfully represented microvascular permeability. Inhibiting endothelial connexin43 (Cx43) gap junction communication with Gap27 blunted the acid-induced reduction in normalized fluorescence, although scrambled Gap27 did not have any effect. The blunting was evident not only in microvessels away from the site of injury, but also in those abutting directly injured alveoli. Thus the new fluorescence-based method reveals that acid increases microvascular permeability both at acid-instilled and away sites. Inhibiting endothelial Cx43 blocked the permeability increase even at the direct injury sites. These data indicate for the first time that Cx43-dependent mechanisms mediate acid-induced increases in microvascular permeability. Cx43 may be a therapeutic target in acid injury.
authors.affiliation: Dept. of Physiology, 894 Union Ave., #426, Memphis, TN 38163, USA. kparthas@uthsc.edu
authors.forename: Kaushik
authors.initials: K
authors.lastname: Parthasarathi
mesh_terms: D000143:Acids / Q000378:metabolism*; D000818:Animals; D002199:Capillary Permeability / Q000502:physiology*; D018031:Connexin 43 / Q000378:metabolism*; D003911:Dextrans / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism / Q000502:physiology*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism / Q000502:physiology; D006851:Hydrochloric Acid / Q000378:metabolism*; D008168:Lung / Q000098:blood supply*; D008297:Male; D011650:Pulmonary Alveoli / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000143:Acids; D018031:Connexin 43; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate; D006851:Hydrochloric Acid