Cardiac tissue iron: effects on post-ischemic function and free radical production, and its possible role during preconditioning.
We determined whether prior treatment of rats (study 1) with subthreshold doses of iron (no evidence of cardiac tissue overload), or in vitro ischemic pre-conditioning (IP: 5 min. Ischemia (I)/5 min. Reperfusion (R) x 2 cycles) of hearts from untreated rats (study 2), can modulate redox-active cardiac tissue iron levels or distribution, leading to alterations in post-ischemic lipid peroxidation-derived free radical (FR) production and severity of reperfusion injury. In study 1, rats received biweekly i.p. injections of 0 (saline=S), 3, 6, or 12 mg FeCl3/ml for 3-wks prior to imposing 30 min. I/15 min. R in vitro. The highest dose caused no elevations in plasma or heart tissue Fe levels, but did further reduce post-ischemic recoveries of left ventricular developed pressure (17% lower), cardiac work (57%) and output (54%), and increased effluent lipid hydroperoxides (2.1-fold) compared to the S-group. Post-ischemic FR production was assessed in toluene-extracted effluent by ESR spectroscopy and alpha-phenyl-N-tert butylnitrone (PBN=2.5 mM perfusate) spin trapping. PBN/alkoxyl (alphaH=1.90 G, alphaN=13.63 G) was the dominant signal detected in all groups; however, Fe-treated groups displayed significant dose-dependent increases in total alkoxyl content (3, 6, 12 mg/ml: 1.8-, 2.3-, 2.7-fold higher) compared to the S-group. These data suggest that even mild, non-overloading doses of iron can be functionally and oxidatively detrimental to hearts when an I/R stress is imposed. In study 2, isolated hearts from untreated rats were exposed to two-IP cycles: during IP, total effluent iron content (atomic absorption) increased 11.4-fold compared to control and analysis of cardiovascular tissue iron distribution (X-ray microanalysis) suggested that iron loss from capillary endothelium was far greater than from tissue myocytes. Moreover, iron-catalyzed production of alkoxyl radicals following severe I/R stress (40 min. I/15 min. R) was substantially lower (73%) in IP hearts compared to the non-IP counterparts. These preliminary findings suggest that cardioprotection resulting from IP may, in part, be related to IP-induced release of cardiovascular endothelial iron (redox-active) prior to imposing severe I/R stress.
Kramer
J H
JH
Department of Physiology and Experimental Medicine, The George Washington University Medical Center, Washington, DC 20037, USA. phyjhk@gwumc.edu
Lightfoot
F G
FG
Weglicki
W B
WB
D000818:Animals; D003497:Cyclic N-Oxides; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D007501:Iron / Q000097:blood / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D007511:Ischemia; D019157:Ischemic Preconditioning, Myocardial; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D009589:Nitrogen Oxides / Q000627:therapeutic use; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D013054:Spectrophotometry, Atomic; D019097:Spin Trapping; D013997:Time Factors
D003497:Cyclic N-Oxides; D005609:Free Radicals; D009589:Nitrogen Oxides; C029217:phenyl-N-tert-butylnitrone; D007501:Iron