Published online before print March 6, 2003, doi: 10.1161/01.RES.0000065167.52922.F6
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© 2003 American Heart Association, Inc.
Integrative Physiology |
12-Lipoxygenase in Opioid-Induced Delayed Cardioprotection
Gene Array, Mass Spectrometric, and Pharmacological Analyses
From the Medical College of Wisconsin, Department of Pharmacology and Toxicology, Milwaukee (H.H.P., E.R.G., A.K.H., M.I., K.N., G.J.G.); Brigham and Women’s Hospital/Harvard Medical School, Center for Neurological Diseases, Cambridge, Mass (R.M.F., S.R.G.); Wesleyan University, Department of Physics, Middletown, Conn (L.O.V.E., R.V.J.); and the University of California, San Diego, Department of Pharmacology, La Jolla (H.H.P., R.A.B., P.A.I.).
Correspondence to Garrett J. Gross, PhD, Medical College of Wisconsin, Department of Pharmacology and Toxicology, 8701 Watertown Plank Rd, Milwaukee, WI 53226. E-mail ggross{at}mcw.edu
12-Lipoxygenase (12-LO) has been shown to be a factor in acute ischemic preconditioning (IPC) in the isolated rat heart; however, no studies have been reported in delayed PC. We characterized the role of 12-LO in an intact rat model of delayed PC induced by a 
-opioid agonist SNC-121 (SNC). Rats were pretreated with SNC and allowed to recover for 24 hours. They were then treated with either baicalein or phenidone, 2 selective 12-LO inhibitors. In addition, SNC-pretreated rats had plasma samples isolated at different times after ischemia-reperfusion for liquid chromatographic–mass spectrometric analysis of the major metabolic product of 12-LO, 12-HETE. Similar studies were conducted with inhibitors. Gene array data showed a significant induction of 12-LO message (P<0.05) after opioid pretreatment. This induction in 12-LO mRNA was confirmed by real-time polymerase chain reaction, and 12-LO protein expression was enhanced by SNC pretreatment at 24 hours relative to vehicle treatment. Both baicalein and phenidone attenuated the protective effects of SNC pretreatment on infarct size (50±4% and 42±3% versus 29±2%, P<0.05, respectively). No significant differences were observed in 12-HETE concentrations between baseline control and SNC-treated rats. However, 12-HETE concentrations were increased significantly at both 15 minutes during ischemia and at 1 hour of reperfusion in the SNC-treated rats compared with controls. Baicalein and phenidone attenuated the increase in 12-HETE at 1 hour of reperfusion. These data suggest that SNC-121 appears to enhance message and subsequently the activity and expression of 12-LO protein during times of stress, resulting in delayed cardioprotection.
Key Words: ischemia • opioids • late phase • gene array • lipid metabolites
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