Role of inducible nitric oxide synthase in induction of RhoA expression in hearts from diabetic rats.

Abstract

AIMS:

Recent studies from our laboratory demonstrated that increased expression of the small GTP-binding protein RhoA and activation of the RhoA/rho kinase (ROCK) pathway play an important role in the contractile dysfunction associated with diabetic cardiomyopathy in hearts from streptozotocin (STZ)-induced diabetic rats. Nitric oxide (NO) has been reported to be a positive regulator of RhoA expression in vascular smooth muscle, and we have previously found that the expression of inducible NO synthase (iNOS) is increased in hearts from STZ-diabetic rats. Therefore, in this study, we investigated the hypothesis that induction of iNOS positively regulates RhoA expression in diabetic rat hearts.

METHODS AND RESULTS:

To determine whether NO and iNOS could increase RhoA expression in the heart, cardiomyocytes from non-diabetic rats were cultured in the presence of the NO donor sodium nitroprusside (SNP) or lipopolysaccharide (LPS) in the absence and presence of the selective iNOS inhibitor, N(6)-(1-iminoethyl)-l-lysine dihydrochloride (L-NIL). In a second study, 1 week after induction of diabetes with STZ, rats were treated with L-NIL (3 mg/kg/day) for 8 more weeks to determine the effect of iNOS inhibition in vivo on RhoA expression and cardiac contractile function. Expression of iNOS was elevated in cardiomyocytes isolated from diabetic rat hearts. Both SNP and LPS increased RhoA expression in non-diabetic cardiomyocytes. The LPS-induced elevation in RhoA expression was accompanied by an increase in iNOS expression and prevented by L-NIL. Treatment of diabetic rats with L-NIL led to a significant improvement in left ventricular developed pressure and rates of contraction and relaxation concomitant with normalization of total cardiac nitrite levels, RhoA expression, and phosphorylation of the ROCK targets LIM (Lin-11, Isl-1, Mec-3) kinase and ezrin/radixin/moesin.

CONCLUSION:

These data suggest that iNOS is involved in the increased expression of RhoA in diabetic hearts and that one of the mechanisms by which iNOS inhibition improves cardiac function is by preventing the upregulation of RhoA and its availability for activation.

• Authors: Hesham Soliman, Graham P Craig, Prabhakar Nagareddy, Violet G Yuen, Guorong Lin, Ujendra Kumar, John H McNeill, Kathleen M Macleod
• Journal: Cardiovascular research (Cardiovasc Res) Vol. 79 Issue 2 Pg. 322-30 (Jul 15 2008) ISSN: 0008-6363 [Print] England
• PMID: 18411229 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Enzyme Inhibitors
• Lipopolysaccharides
• N(6)-(1-iminoethyl)lysine
• Nitric Oxide Donors
• Nitroprusside
• Streptozocin
• Nitric Oxide Synthase Type II
• rho-Associated Kinases
• rhoA GTP-Binding Protein
• Lysine

Topics

• Animals
• Cells, Cultured
• Diabetes Mellitus, Experimental (metabolism, pathology)
• Disease Models, Animal
• Dose-Response Relationship, Drug
• Enzyme Inhibitors (pharmacology)
• Lipopolysaccharides (pharmacology)
• Lysine (analogs & derivatives, pharmacology)
• Male
• Myocardial Contraction (drug effects)
• Myocardium (metabolism, pathology)
• Myocytes, Cardiac (drug effects, metabolism, pathology)
• Nitric Oxide Donors (pharmacology)
• Nitric Oxide Synthase Type II (metabolism)
• Nitroprusside (pharmacology)
• Rats
• Streptozocin
• rho-Associated Kinases (metabolism)
• rhoA GTP-Binding Protein (metabolism)