Induction of iNOS gene expression by monophosphoryl lipid A: a pharmacological approach for myocardial adaptation to ischemia.
Using the concept that exposing a cell to an adverse environment (stress) results in the stimulation of its endogenous defense system, hearts have been adapted to ischemia by exposing them to diverse stresses. Recently, 24-h pretreatment of monophosphoryl lipid A (MLA), a chemically modified derivative of endotoxin, was found to render the hearts more tolerant to ischemic reperfusion injury. Since nitric oxide has recently been implicated in myocardial preservation and since inducible nitric oxide synthetase (iNOS) was originally characterized in macrophages and shown to be maximally induced by bacterial lipopolysaccharides (endotoxin), we sought to determine whether MLA mediates its cardioprotective effects through the iNOS expression. For this, rats were injected with MLA (300 micrograms/kg) or vehicle (control), and after 24 h the animals were sacrificed and the isolated working hearts were made ischemic for 30 min followed by 30 min of reperfusion. MLA-treated hearts were found to be tolerant to ischemic reperfusion injury as evidenced by improved postischemic ventricular recovery. After 30 min of reperfusion, left ventricular developed pressure (LVDP) and its maximum first derivative (LVmaxdp/dt) were 13.3 +/- 0.3 kPa and 537 +/- 13 kPa/s, respectively, in the MLA-treated group, as compared with 10.2 +/- 0.4 kPa (p < 0.05) and 447 +/- 11 kPa/s (p < 0.05), respectively, for the control group. Aortic flow and coronary flow were 20.1 +/- 1.4 ml/min and 19.1 +/- 0.8 ml/min, respectively, in the MLA group, as compared with 9.5 +/- 0.8 ml/min (p < 0.05) and 15.9 +/- 0.7 ml/min (p < 0.05), respectively, for the untreated group. 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 h of treatment) and Northern blot analysis was performed using specific cDNA probe for iNOS. A single band of approximately 4.6 kb corresponding to iNOS mRNA was detected after 4 h of MLA treatment, while the maximal iNOS expression was found between 6-8 h of MLA treatment. The results of this study demonstrate that MLA induces the expression of iNOS and protects the myocardium from ischemic reperfusion injury.
Maulik
N
N
Department of Surgery, University of Connecticut School of Medicine, Farmington 06030-1110, USA.
Tosaki
A
A
Elliott
G T
GT
Maulik
G
G
Das
D K
DK
D000818:Animals; D015152:Blotting, Northern; D015971:Gene Expression Regulation, Enzymologic; D008050:Lipid A / Q000031:analogs & derivatives* / Q000494:pharmacology; D008297:Male; D017202:Myocardial Ischemia / Q000378:metabolism / Q000503:physiopathology; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000517:prevention & control*
D008050:Lipid A; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; C048436:monophosphoryl lipid A