Tuning the nitric oxide release from CPO-27 MOFs.

Nitric oxide (NO) storage and release measurements have been recorded for Ni-doped CPO-27 (Mg) and CPO-27 (Zn), and the biological effect of the released NO was assessed in porcine coronary artery relaxation tests. The results indicate that the doping strategy leads to increased levels of NO storage and delivery compared to the parent materials and that the NO dosage and biological response can be tuned via this approach to suit the requirements of particular applications.

Maternal fat intake in rats alters 20:4n-6 and 22:6n-3 status and the epigenetic regulation of Fads2 in offspring liver.

Poor prenatal nutrition, acting through epigenetic processes, induces persistent changes in offspring phenotype. We investigated the effect of maternal fat intake on polyunsaturated fatty acid (PUFA) status and on the epigenetic regulation of Fads2, encoding Δ6 desaturase (rate limiting in PUFA synthesis), in the adult offspring. Rats (n=6 per dietary group) were fed either 3.5% (w/w), 7% (w/w) or 21% (w/w) butter or fish oil (FO) from 14 days preconception until weaning. Offspring (n=6 males and females per dietary group) were fed 4% (w/w) soybean oil until postnatal day 77. 20:4n-6 and 22:6n-3 levels were lower in liver phosphatidylcholine (PC) and phosphatidylethanolamine and plasma PC (all P<.0001) in offspring of dams fed 21% than 3.5% or 7% fat regardless of type. Hepatic Fads2 expression related inversely to maternal dietary fat. Fads2 messenger RNA expression correlated negatively with methylation of CpGs at -623, -394, -84 and -76 bases relative to the transcription start site (all P<.005). Methylation of these CpGs was higher in offspring of dams fed 21% than 3.5% or 7% fat; FO higher than butter. Feeding adult female rats 7% fat reduced 20:4n-6 status in liver PC and Fads2 expression and increased methylation of CpGs -623, -394, -84 and -76 that reversed in animals switched from 7% to 4% fat diets. These findings suggest that fat exposure during development induces persistent changes, while adults exhibit a transient response, in hepatic PUFA status in offspring through epigenetic regulation of Fads2. Thus, epigenetic regulation of Fads2 may contribute to short- and long-term regulation of PUFA synthesis.

Vascular dysfunction induced in offspring by maternal dietary fat involves altered arterial polyunsaturated fatty acid biosynthesis.

Nutrition during development affects risk of future cardiovascular disease. Relatively little is known about whether the amount and type of fat in the maternal diet affect vascular function in the offspring. To investigate this, pregnant and lactating rats were fed either 7%(w/w) or 21%(w/w) fat enriched in either 18:2n-6, trans fatty acids, saturated fatty acids, or fish oil. Their offspring were fed 4%(w/w) soybean oil from weaning until day 77. Type and amount of maternal dietary fat altered acetylcholine (ACh)-mediated vaso-relaxation in offspring aortae and mesenteric arteries, contingent on sex. Amount, but not type, of maternal dietary fat altered phenylephrine (Pe)-induced vasoconstriction in these arteries. Maternal 21% fat diet decreased 20:4n-6 concentration in offspring aortae. We investigated the role of Δ6 and Δ5 desaturases, showing that their inhibition in aortae and mesenteric arteries reduced vasoconstriction, but not vaso-relaxation, and the synthesis of specific pro-constriction eicosanoids. Removal of the aortic endothelium did not alter the effect of inhibition of Δ6 and Δ5 desaturases on Pe-mediated vasoconstriction. Thus arterial smooth muscle 20:4n-6 biosynthesis de novo appears to be important for Pe-mediated vasoconstriction. Next we studied genes encoding these desaturases, finding that maternal 21% fat reduced Fads2 mRNA expression and increased Fads1 in offspring aortae, indicating dysregulation of 20:4n-6 biosynthesis. Methylation at CpG -394 bp 5' to the Fads2 transcription start site predicted its expression. This locus was hypermethylated in offspring of dams fed 21% fat. Pe treatment of aortae for 10 minutes increased Fads2, but not Fads1, mRNA expression (76%; P<0.05). This suggests that Fads2 may be an immediate early gene in the response of aortae to Pe. Thus both amount and type of maternal dietary fat induce altered regulation of vascular tone in offspring though differential effects on vaso-relaxation, and persistent changes in vasoconstriction via epigenetic processes controlling arterial polyunsaturated fatty acid biosynthesis.

Atorvastatin restores endothelial function in offspring of protein-restricted rats in a cholesterol-independent manner.

Maternal protein restriction in rats leads to endothelial dysfunction and decreased NO bioavailability in the offspring. Statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) are recognized to have pleiotropic actions including increasing NO bioavailability and reducing inflammation and oxidative damage. This study assessed statin treatment on vascular function in a model of endothelial dysfunction, which is independent of dyslipidemia. Wistar rats were fed a control (18% casein) or protein-restricted (9% casein) diet throughout pregnancy. At weaning, a subset of the protein-restricted group was given atorvastatin (10 mg/kg per day) in the drinking water. At 145 days of age, offspring were euthanized by CO(2) inhalation. Plasma samples were collected for markers of inflammation, vascular reactivity of the thoracic aorta, and small mesenteric arteries were assessed on the wire myograph, and tissues were snap frozen for molecular biology analysis. Thoracic aorta endothelial-dependent vasodilatation was attenuated in the male offspring from both protein-restricted groups compared with controls (P<0.05) but was similar in females (P value not significant). Endothelial-dependent dilatation of mesenteric arteries was attenuated in male and female protein-restricted offspring (P<0.05) and was corrected by atorvastatin. Maternal protein restriction increased plasma inflammatory markers granulocyte chemotactic protein, lipocalin-2, and beta(2)-microglobulin in male and C-reactive protein in female offspring (P<0.05). Atorvastatin had no effect on inflammatory markers in the males but restored C-reactive protein to control levels in the females (P<0.05). Aortic and mesenteric artery mRNA levels of endothelial NO synthase, superoxide dismutase 1, and tumor necrosis factor-alpha were unchanged. These data suggest that atorvastatin can restore endothelial function in this model, but its effects are gender specific and dependent on the vascular bed.

Expression and localization of C-type natriuretic peptide in human vascular smooth muscle cells.

OBJECTIVES: C-type natriuretic peptide (CNP) released by vascular endothelium relaxes smooth muscle and is important in the maintenance of vascular tone. Since it is not known whether other human vascular cell types produce CNP, we investigated its expression in human vascular smooth muscle. METHODS: CNP expression was examined by RT-PCR in vascular smooth muscle cells (SMC) cultured from human saphenous vein (SV), internal mammary artery (IMA) and radial artery (RA), and CNP protein was probed using immunostaining, in tissue sections and in SMCs cultured from these vessels, respectively. RESULTS: PCR for CNP produced a 334 bp product in all SMC cultures, as expressed in endothelial cells, although the band intensity was markedly less in SMCs. Myocardium from CNP-knockout mouse did not express CNP, while there was expression in wild-type mouse. CNP protein was detected by immunostaining in 100% of SMC cultures. By immunostaining of tissue sections, CNP was detected throughout the medial layer, but not adventitia, of all vessel types. CONCLUSIONS: Expression of CNP at gene and protein level by human vascular SMCs suggests that CNP may have the capacity to regulate vascular tone independently of the endothelium.

C-type natriuretic peptide relaxes human coronary artery bypass grafts preconstricted by endothelin-1.

BACKGROUND: Endothelin is implicated in graft spasm after coronary artery bypass grafting. We assessed reversal by the endothelium-derived vasodilator C-type natriuretic peptide of prior contraction of radial artery and other vessels commonly used for coronary artery bypass surgery. METHODS: Segments of human radial artery, saphenous vein, and internal mammary artery were mounted in organ baths after removal from patients undergoing cardiac surgery (n = 34; 64 +/- 2 years). Effects of increasing concentrations of C-type natriuretic peptide (with or without aprotinin, 1,000 U/mL) on endothelin-induced contraction were compared with acetylcholine, sodium nitroprusside, and papaverine. RESULTS: C-type natriuretic peptide relaxed endothelin precontraction in all vessels (F = 17.8, 36.3, and 48.4, respectively; p < 0.001), with maximum relaxations of 44%, 54%, and 66% in saphenous vein, internal mammary artery, and radial artery, respectively. Aprotinin did not affect relaxation to C-type natriuretic peptide. Acetylcholine relaxed the saphenous vein weakly, with maximal relaxation of 9% at 10(-6) M. However, the radial artery and internal mammary artery relaxed strongly to acetylcholine. The highest concentration of papaverine completely relaxed all vessels, but responses were less sensitive than to sodium nitroprusside or acetylcholine. CONCLUSIONS: C-type natriuretic peptide reverses endothelin-induced constriction in arterial and venous conduits used for coronary artery bypass, particularly the radial artery. Proteolytic breakdown of C-type natriuretic peptide by local vascular enzymes appears of little importance in vitro. This signals the therapeutic potential of using C-type natriuretic peptide as an antagonist of graft vasospasm after coronary artery bypass surgery.