Cardioprotection by hypoxia-inducible factor 1 alpha transfection in skeletal muscle is dependent on haem oxygenase activity in mice.
OBJECTIVE
The present study investigates whether the cardioprotection achieved by gene delivery of hypoxia-inducible factor-1 alpha (HIF-1 alpha) depends on the downstream factor haem oxygenase (HMOX)-1.


RESULTS
Immortalized cardiomyocytes (HL-1 cells) were transfected with HIF-1 alpha or HMOX-1 and injured with hydrogen peroxide (H(2)O(2)), and death was evaluated by trypan blue staining. Quadriceps muscles of mice were treated with DNA for HIF-1 alpha and HMOX-1, or sham-treated and electroporated, and 3 days later, hearts were isolated and subjected to global ischaemia and reperfusion. Some HIF-1 alpha- and sham-treated mice received the HMOX blocker zinc deuteroporphyrin 2,4-bis-glycol (ZnBG) (n = 6-8 in each group). HL-1 cells were stimulated with bilirubin or the carbon monoxide donor CORM-2 before injury with H(2)O(2). HL-1 cells which were transfected with HIF-1 alpha or HMOX-1 had an increased survival to H(2)O(2)-induced injury compared with empty vector (n = 10-12 per group; P < 0.01 for both). When HMOX-1-luciferase reporter mice were treated with HIF-1 alpha in the quadriceps muscle, increased luciferase activity was found locally, but nowhere else. Mice pre-treated with HIF-1 alpha or HMOX-1 had a reduced infarct size, improved post-ischaemic function, and increased serum bilirubin (P < 0.05). ZnBG inhibited all these effects afforded by HIF-1 alpha. Stimulation of HL-1 cells with bilirubin and CORM-2 reduced cell death evoked by H(2)O(2) (P < 0.05 for both, n = 11-15 in each group).


CONCLUSIONS
HIF-1 alpha and HMOX-1 provided protection against H(2)O(2)-induced damage in HL-1 cells. Remote gene delivery of HIF-1 alpha afforded cardioprotective effects. These were dependent on HMOX activity, as an HMOX blocker abolished the effects, and they were mimicked by pre-treatment with HMOX-1. Downstream to HMOX-1, bilirubin as well as carbon monoxide may be organ effectors.
Czibik
Gabor
G
Department of Physiology, IMB, Institute of Basic Medical Sciences, University of Oslo, PO Box 1103, Blindern, NO-0317 Oslo, Norway. gaborc@medisin.uio.no
Sagave
Julia
J
Martinov
Vladimir
V
Ishaq
Bushra
B
Sohl
Marcus
M
Sefland
Iren
I
Carlsen
Harald
H
Farnebo
Filip
F
Blomhoff
Rune
R
Valen
Guro
G
D000818:Animals; D001663:Bilirubin / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D003905:Deuteroporphyrins / Q000494:pharmacology; D004195:Disease Models, Animal; D018274:Electroporation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015316:Genetic Therapy; D051547:Heme Oxygenase-1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009203:Myocardial Infarction / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D015428:Myocardial Reperfusion Injury / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D009942:Organometallic Compounds / Q000494:pharmacology; D016877:Oxidants / Q000633:toxicity; D052097:Quadriceps Muscle / Q000187:drug effects / Q000201:enzymology*; D013997:Time Factors; D014162:Transfection; D016277:Ventricular Function, Left; D017725:Ventricular Pressure
D003905:Deuteroporphyrins; D004791:Enzyme Inhibitors; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008565:Membrane Proteins; D009942:Organometallic Compounds; D016877:Oxidants; C447082:tricarbonyldichlororuthenium (II) dimer; C060750:zinc deuteroporphyrin IX 2,4-bis(glycol); D006861:Hydrogen Peroxide; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D001663:Bilirubin