TY - JOUR
T1 - Protective effect of heme oxygenase-1 gene transfer against oxyhemoglobin-induced endothelial dysfunction
AU - Eguchi, Daihiko
AU - Weiler, Deborah
AU - Alam, Jawed
AU - Nath, Karl
AU - Katusic, Zvonimir S.
PY - 2001
Y1 - 2001
N2 - The current study was designed to determine the effect of recombinant heme oxygenase-1 (HO-1) gene expression on endothelial function in cerebral arteries. Isolated canine basilar arteries were exposed ex vivo (30 minutes at 37°C) to an adenoviral vector (1010 PFU/mL, total volume 300 μL) encoding either the HO-1 gene (AdCMVHO-1) or the β-galactosidase (β-Gal) reporter gene (AdCMVβ-Gal). Twenty-four hours after transduction, arterial rings were suspended in organ chamber for isometric force recording. Endothelium-dependent relaxations were obtained in response to bradykinin (10-10 to 10-6 mol/L) during contraction to uridine-5′-triphosphate (UTP; 3 x 10-6 to 3 x 10-5 mol/L). Certain rings were incubated with oxyhemoglobin (OxyHb; 10-5 mol/L) overnight (16 to 18 hours of 24 hours). Expression and localization of recombinant protein were shown by Western blot analysis and immunohistochemistry. Endothelium-dependent relaxation to bradykinin and endothelium-independent relaxation to forskolin (10-9 to 10-5 mol/L) and DEA-NONOate (10-10 to 10-5 mol/L) were identical in β-Gal- and HO-1-transduced arteries. Exposure to OxyHb caused impairment of endothelium-dependent relaxation to bradykinin (P < 0.01). In contrast, OxyHb did not affect endothelium-dependent relaxation in arteries expressing recombinant HO-1 (P > 0.05). This protective effect of HO-1 was reversed by coincubation with tin protoporphyrin (SnPP9; 10-5 mol/L), a selective inhibitor of HO-1 (P < 0.01). Basal levels of 3′,5′-cyclic monophosphate (cGMP) in HO-1-transduced vessels were not significantly different from those in β-Gal-transduced vessels. Pretreatment with OxyHb significantly reduced cGMP level in β-Gal-transduced rings (P < 0.01), whereas it had no effect in HO-1-transduced rings. These results demonstrate that HO-1 gene transfer does not affect endothelial and smooth muscle function of normal arteries, and that expression of recombinant HO-1 in cerebral arteries protects vasomotor function against OxyHb-induced injury.
AB - The current study was designed to determine the effect of recombinant heme oxygenase-1 (HO-1) gene expression on endothelial function in cerebral arteries. Isolated canine basilar arteries were exposed ex vivo (30 minutes at 37°C) to an adenoviral vector (1010 PFU/mL, total volume 300 μL) encoding either the HO-1 gene (AdCMVHO-1) or the β-galactosidase (β-Gal) reporter gene (AdCMVβ-Gal). Twenty-four hours after transduction, arterial rings were suspended in organ chamber for isometric force recording. Endothelium-dependent relaxations were obtained in response to bradykinin (10-10 to 10-6 mol/L) during contraction to uridine-5′-triphosphate (UTP; 3 x 10-6 to 3 x 10-5 mol/L). Certain rings were incubated with oxyhemoglobin (OxyHb; 10-5 mol/L) overnight (16 to 18 hours of 24 hours). Expression and localization of recombinant protein were shown by Western blot analysis and immunohistochemistry. Endothelium-dependent relaxation to bradykinin and endothelium-independent relaxation to forskolin (10-9 to 10-5 mol/L) and DEA-NONOate (10-10 to 10-5 mol/L) were identical in β-Gal- and HO-1-transduced arteries. Exposure to OxyHb caused impairment of endothelium-dependent relaxation to bradykinin (P < 0.01). In contrast, OxyHb did not affect endothelium-dependent relaxation in arteries expressing recombinant HO-1 (P > 0.05). This protective effect of HO-1 was reversed by coincubation with tin protoporphyrin (SnPP9; 10-5 mol/L), a selective inhibitor of HO-1 (P < 0.01). Basal levels of 3′,5′-cyclic monophosphate (cGMP) in HO-1-transduced vessels were not significantly different from those in β-Gal-transduced vessels. Pretreatment with OxyHb significantly reduced cGMP level in β-Gal-transduced rings (P < 0.01), whereas it had no effect in HO-1-transduced rings. These results demonstrate that HO-1 gene transfer does not affect endothelial and smooth muscle function of normal arteries, and that expression of recombinant HO-1 in cerebral arteries protects vasomotor function against OxyHb-induced injury.
KW - Canine basilar artery
KW - Gene transfer
KW - Heme oxygenase
KW - Oxyhemoglobin
KW - SnPP9
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U2 - 10.1097/00004647-200110000-00010
DO - 10.1097/00004647-200110000-00010
M3 - Article
C2 - 11598499
AN - SCOPUS:0034774790
SN - 0271-678X
VL - 21
SP - 1215
EP - 1222
JO - Journal of Cerebral Blood Flow and Metabolism
JF - Journal of Cerebral Blood Flow and Metabolism
IS - 10
ER -