Modulation of corneal heme oxygenase expression by oxidative stress agents.
Heme oxygenase, the rate-limiting enzyme in the degradation of heme to bile-pigments and carbon monoxide, is induced in response to increased oxidative stress and is believed to provide a cytoprotective effect. We investigated the role of heme oxygenase in cultured rabbit corneal epithelial cells (RCE), and its potential to alleviate oxidative stress-induced cell damage. Heme oxygenase in RCE was effectively and potently induced by most metals tested, including tin, silver, and gold, and cytokines such as IL-6, and TGF beta. Stannous chloride and heme-induced heme oxygenase mRNA by 40 and 100 fold within 1-3 hours and increased enzyme activity by 9.2- and 10-fold, respectively, over a 24 hour period. IL-6, TGF beta and H2O2 induced heme oxygenase by 2-3 fold. Zinc protoporphyrins were effective inhibitors of heme oxygenase activity in vitro. However, when incubated with cells for 24 h they induced heme oxygenase mRNA but decreased or had no effect on its activity. Administration of heme, SnCl2, and H2O2 resulted in some degree of glutathione perturbation (GSH/GSSG). However, in all cases, depletion of glutathione was exacerbated if heme oxygenase was simultaneously inhibited. Conversely, perturbation of glutathione levels was minimized if heme oxygenase was induced by heme or stannous chloride. These results demonstrate that RCE cells exhibit functional heme oxygenase activity which is inducible in response to inflammatory cytokines and oxidative stress agents and suggest a cytoprotective role for heme oxygenase against cell injury.
Neil
T K
TK
Rockefeller University, New York, New York, USA.
Stoltz
R A
RA
Jiang
S
S
Laniado-Schwartzman
M
M
Dunn
M W
MW
Levere
R D
RD
Kappas
A
A
Abraham
N G
NG
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015152:Blotting, Northern; D002460:Cell Line; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D016207:Cytokines / Q000494:pharmacology*; D004790:Enzyme Induction; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D006418:Heme / Q000378:metabolism; D006419:Heme Oxygenase (Decyclizing) / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D008670:Metals / Q000494:pharmacology*; D018384:Oxidative Stress; D011524:Protoporphyrins / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D014805:Vitamin B 12 / Q000494:pharmacology
D000975:Antioxidants; D016207:Cytokines; D004791:Enzyme Inhibitors; D008670:Metals; D011524:Protoporphyrins; D012333:RNA, Messenger; C017803:zinc protoporphyrin; D006418:Heme; D006419:Heme Oxygenase (Decyclizing); D005978:Glutathione; D014805:Vitamin B 12; D019803:Glutathione Disulfide