Pharmacological opening of mitochondrial cardiac ATP-sensitive potassium (K ATP ) channels has the chance to be a promising but still controversial cardioprotective mechanism. Physiological roles of mitochondrial K ATP channels in the myocardium remain unclear. We studied the effects of diazoxide, a specific opener of these channels, on the function of rat mitochondria in situ in saponin-permeabilized fibers using an ionic medium that mimics the cytosol. In the presence of NADHproducing substrates (malate ؉ glutamate), neither 100 M diazoxide nor 100 M glibenclamide (a K ATP channel blocker) changed the mitochondrial respiration in the absence or presence of ADP. Because the K ATP channel function could be modified by changes in adenine nucleotide concentrations near the mitochondria, we studied the effects of diazoxide and glibenclamide on the functional activity of mitochondrial kinases. Both diazoxide and glibenclamide did not change the in situ ADP sensitivity in the presence or absence of creatine (apparent K m values for ADP were, respectively, 59 ؎ 9 and 379 ؎ 45 M). Similarly, stimulation of the mitochondrial respiration with AMP in the presence of ATP due to adenylate kinase activity was not affected by the modulators of K ATP channels. However, when succinate was used as substrate, diazoxide significantly inhibited basal respiration by 22% and maximal respiration by 24%. Thus, at a cardioprotective dose, the main functional effect of diazoxide depends on respiratory substrates and seems not to be related to K ATP channel activity.

.-The effect of docosahexaenoic acid (DHA) intake on cardiac mitochondrial function was evaluated in permeabilized fibers in insulin deficiency and insulin resistance in rats. The insulin-deficient state was obtained by streptozotocin injection 2 mo before investigations. Insulin resistance was obtained by feeding a 62% fructose diet for 3 mo. DHA was incorporated in the diet to modify the fatty acid composition of cardiac membranes, including mitochondria. Insulin deficiency decreased mitochondrial creatine kinase (mi-CK) activity and mitochondrial sensitivity to ADP. DHA intake prevented these alterations. Moreover, the insulin-deficient state significantly decreased n-3 polyunsaturated fatty acids (PUFA) and slightly increased n-6 PUFA in both cardiac and mitochondrial membranes, inducing a significant increase in the n-6-to-n-3 ratio. DHA intake maintained high myocardial and mitochondrial DHA content. Insulin deficiency also decreased glutamate-and palmitoylcarnitine-supported mitochondrial respiration, but DHA intake did not prevent these effects. In contrast, insulin resistance did not affect mi-CK activity or sensitivity to ADP. However, insulin resistance influenced the myocardial fatty acid composition with decreased n-6 and n-3 PUFA contents and increased monounsaturated fatty acid content. Only slight alterations were observed in mitochondrial fatty acid composition, and they were corrected by DHA intake. Moreover, insulin resistance decreased the glutamatesupported respiration, and DHA intake did not influence this effect. In conclusion, the impairment of cardiac mitochondrial function was more pronounced in the insulin-deficient state than in insulin resistance. The modification of fatty acid composition of cardiac and mitochondrial membranes by DHA partially prevented the mitochondrial alterations induced in the two models. mitochondrial respiration; myocardium; polyunsaturated fatty acid DIABETES IS ASSOCIATED WITH high cardiovascular morbidity and mortality, and both insulin-deficient and insulin-resistant diabetes induce biochemical and physiological changes in the heart (15). Animal studies have demonstrated a number of diabetes-induced changes in the diabetic heart, including changes in mitochondrial function (14) and fatty acid composition of heart membrane phospholipids. In uncontrolled diabetes, myocardial glucose utilization is markedly reduced, causing fatty acids to support a greater portion of total energy metabolism (23). Moreover, when diabetic rat hearts are perfused in the absence of fatty acids, glucose oxidation represents Ͻ20% of the ATP synthesis (14), suggesting an alteration of the fatty acid uptake regulation in mitochondria. In the diabetic heart, the level of malonyl-CoA, the physiological inhibitor of carnitine palmitoyltransferase I, seems to be reduced. In streptozotocin (STZ) diabetic rats, the activity of acetyl-CoA carboxylase, which produces malonyl-CoA, was shown to decrease (14). In the same model, Sakamoto et al. (24) reported an increase in malonyl-CoA decarboxy...

1. The aim of the present study was to investigate the responses to acetylcholine (ACh; 3 nmol/L-30 micromol/L) and sodium nitroprusside (SNP; 3 nmol/L-30 micromol/L) of precontracted aortic rings from diabetic rats supplemented with docosahexaenoic acid (DHA). 2. Diabetes was induced by streptozotocin (STZ; 55 mg/kg). Diabetic and sham rats were fed, over a period of 8 weeks, either control diet or a DHA-supplemented diet. Aortic endothelial fatty acid composition was analysed by gas chromatography. The involvement of endothelial-derived nitric oxide (NO) and cyclo-oxygenase (COX) metabolites in response to ACh was assessed using the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (100 micromol/L) and the COX inhibitor indomethacin (1 micromol/L), respectively. 3. The DHA-supplemented diet induced a small increase in n-3 polyunsaturated fatty acids (PUFA; P < 0.001) owing to the incorporation of DHA in the endothelial cells of sham animals (1.6 +/- 0.2% in the DHA group compared with traces in the control group; P < 0.001) and diabetic animals (1.3 +/- 0.2% in the DHA group compared with traces in control group; P < 0.001), without a decrease in n-6 PUFA, despite a small decrease in arachidonic acid content (P < 0.05). Diabetes did not modify the incorporation of DHA in endothelial cells, but did significantly increase the arachidonic acid content (0.6 +/- 0.0 vs 0.4 +/- 0.1% in control group in the STZ and sham groups, respectively; P < 0.001). Acetylcholine-induced relaxation was significantly reduced in STZ groups compared with the sham groups (P < 0.001) and the DHA-supplemented diet did not modify these effects. In contrast, neither the DHA-supplemented diet nor diabetes affected the aortic relaxation induced by SNP. N(G)-Nitro-L-arginine methyl ester strongly inhibited the relaxant effects of ACh in the sham groups (P < 0.001) and abolished ACh-induced relaxation in the STZ groups (P < 0.001). The diet did not modify these effects. In the presence of indomethacin, the relaxation induced by ACh was decreased in the sham groups (P < 0.01), but not in the STZ groups. The DHA-supplemented diet did not have any effect on these responses. 4. In conclusion, these results suggest that, in the present study, the endothelial dysfunction occurring in the rat model of STZ-induced diabetes is associated with modifications of both the synthesis of COX derivatives and NO metabolism and is not affected by dietary supplementation with DHA.