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Vol. 285, Issue 3, 1274-1279, June 1998
Cardiovascular Division, Department of Surgery, University of
Connecticut School of Medicine, Farmington, Connecticut (A.T., N.M,
D.K.D.),
RIBI ImmunoChem Research, Inc., Hamilton, Montana (G.T.E.),
Institute for Molecular Pharmacology, Berlin, Germany (I.E.B.), and
Department of Surgery, Baystate Medical Center, Springfield,
Massachusetts (R.M.E.)
Preconditioning with monophosphoryl lipid A (MLA) protects rabbit
hearts from prolonged ischemic reperfusion injury by a mechanism involving inducible nitric oxide synthase (iNOS) activation. This study
was undertaken to determine whether MLA also could precondition rat
hearts in a similar manner. Rats were injected with two different doses
of MLA (300 µg/kg or 450 µg/kg i.v.) or vehicle (control), and
after 24 hr the animals were sacrificed for preparation of isolated
perfused rat hearts. Hearts were then perfused by working mode, and
then made ischemic for 30 min followed by 30 min of reperfusion.
Another group of hearts were treated simultaneously with a nitric oxide
(NO) blocker, L-nitro-arginine-methyl-ester (L-NAME) (10 mg/kg) and MLA (450 µg/kg). For arrhythmia
studies, 12 hearts were used in each group (total, 48 hearts). Cardiac functions were examined in a separate group of 24 hearts
(n = 6/group). MLA-treated hearts (either dose)
were tolerant to ischemic reperfusion injury as evidenced by improved
postischemic ventricular recovery [coronary flow (ml/min) 19.1 ± 0.8 (300 µg/kg MLA), 22.6 ± 1.0 (450 µg/kg MLA)
vs. 15.9 ± 0.7 (control); aortic flow (ml/min) 20.7 ± 1.8 (300 µg/kg MLA), 25.8 ± 1.4 (450 µg/kg MLA)
vs. 11.0 ± 0.8 (control); left ventricular
developed pressure (kPa) 13.3 ± 0.6 (300 µg/kg MLA), 14.6 ± 0.2 (450 µg/kg MLA) vs. 10.3 ± 0.7 (control)]. Incidences of ventricular fibrillation and ventricular tachycardia were decreased compared with the control group only in the
450 µg/kg dose of MLA-treated hearts (92% to 33%). Pretreatment of
the hearts with L-NAME inhibited the preconditioning effect of MLA. To examine the induction of the iNOS expression, RNAs were
extracted from the control and MLA-treated hearts (after 2, 4,6, 8, 12 and 24 hr of treatment) and Northern blot analyses were performed with
a specific cDNA probe for iNOS. A single band of approximately 4.6 kb
corresponding to iNOS mRNA was detected after 4 hr of MLA treatment,
whereas the maximal iNOS expression was found between 6 and 8 hr of MLA
treatment. The results of this study demonstrated that MLA induced the
expression of iNOS and protected the myocardium from ischemic
reperfusion injury which is blocked by an inhibitor of NO synthesis,
which suggests a role of NO in MLA-mediated cardioprotection.
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