Fetal growth needs adequate blood perfusion from both sides of the placenta, on the maternal side through the uterine vessels and on the fetal side through the umbilical cord. In a model of intrauterine growth restriction (IUGR) induced by reduced blood volume expansion, uterine artery remodeling was blunted. The aim of this study is to determine if IUGR and fetus sex alter the functional and mechanical parameters of umbilical cord blood vessels. Pregnant rats were given a low sodium (IUGR) or a control diet for the last 7 days of pregnancy. Umbilical arteries and veins from term (22 day) fetal rats were isolated and set-up in wire myographs. Myogenic tone, diameter, length tension curve and contractile response to thromboxane analog U46619 and serotonin (5-HT) were measured. In arteries from IUGR fetuses, myogenic tone was increased in both sexes while diameter was significantly greater only in male fetuses. In umbilical arteries collected from the control group, the maximal contraction to U46619 was lower in females than males. Compared to the control groups, the maximal response decreased in IUGR male arteries and increased in female ones, thus abolishing the sexual dimorphism observed in the control groups. Reduced contractile response to U46619 was observed in the IUGR vein of both sexes. No difference between groups was observed in response to 5HT in arteries. In conclusion, the change in parameters of the umbilical cord blood vessels in response to a mild insult seems to show adaptation that favors better exchange of deoxygenated and wasted blood from the fetus to the placenta with increased myogenic tone.

Context Intrauterine growth restriction (IUGR) is an immediate outcome of an adverse womb environment, exposing newborns to developing cardiometabolic disorders later in life. Objective This study investigates the cardiac metabolic consequences and underlying mechanism of energy expenditure in developing fetuses under conditions of IUGR. Methods Using an animal model of IUGR characterized by uteroplacental vascular insufficiency, mitochondrial function, gene profiling, lipidomic analysis, and transcriptional assay were determined in fetal cardiac tissue and cardiomyocytes. Results IUGR fetuses exhibited an upregulation of key genes associated with fatty acid breakdown and beta-oxidation (Acadvl, Acadl, Acaa2), and mitochondrial carnitine shuttle (Cpt1a, Cpt2), instigating a metabolic gene reprogramming in the heart. Induction of Ech1, Acox1, Acox3, Acsl1, and Pex11a indicated a coordinated interplay with peroxisomal beta-oxidation and biogenesis mainly observed in females, suggesting sexual dimorphism in peroxisomal activation. Concurring with the sex-related changes, mitochondrial respiration rates were stronger in IUGR female fetal cardiomyocytes, accounting for enhanced adenosine 5 '-triphosphate production. Mitochondrial biogenesis was induced in fetal hearts with elevated expression of Ppargc1a transcript specifically in IUGR females. Lipidomic analysis identified the accumulation of arachidonic, eicosapentaenoic, and docosapentaenoic polyunsaturated long-chain fatty acids (LCFAs) in IUGR fetal hearts, which leads to nuclear receptor peroxisome proliferator-activated receptor alpha (PPAR alpha) transcriptional activation in cardiomyocytes. Also, the enrichment of H3K27ac chromatin marks to PPAR alpha-responsive metabolic genes in IUGR fetal hearts outlines an epigenetic control in the early metabolic energy switch. Conclusion This study describes a premature and sex-related remodeling of cardiac metabolism in response to an unfavorable intrauterine environment, with specific LCFAs that may serve as predictive effectors leading to IUGR.

Adverse uterine environment may expose newborns to develop metabolic diseases later in their life. The in-utero growth restriction (IUGR) is an immediate outcome of a sub-optimal womb environment, where the fetus cannot reach its full growth potential. We have developed an animal model of IUGR based on a low-sodium diet given to dams during the last week of gestation. We showed that the isocaloric low-sodium diet reduces maternal blood volume expansion, causing poor placental perfusion, low placental weight and subsequent IUGR in offspring. Redistribution of blood flow in the fetus was observed, causing a preferential perfusion to major organs, such as the heart. Consistent with this, adult offspring showed an increase in blood pressure and remodeling of cardiomyocytes occurs in adult female, suggesting metabolic changes in cardiac function. In this study, we address whether the IUGR induced-stress leads to metabolic changes in fetal cardiomyocytes to adapt their energy-production process and lipid utilization. At 22 days of gestation, hearts were collected from IUGR and control fetuses, and homogenized. Gene expression profile as well as protein levels were analyzed. We observed an increased expression of critical genes involved in lipid metabolism, such as long chain fatty acid receptor CD36, carnitine palmitoyltransferase CPT1a, and also key enzymes of long chain fatty acid beta-oxidation in IUGR heart. In addition, our results indicate an increase in mitochondrial biogenesis. To address whether changes in lipid metabolite profile might prevail, we performed lipidomic MS analysis in heart homogenates. Lipidomics showed significant accumulation of very long and long-chain fatty acids in the context of IUGR and when comparing males and females. These changes could be reminiscent of an adaptation in the use of fatty acids for energy production. Altogether, these findings suggest that fetal IUGR cardiac cells may rely more on the use of lipids as an energy source, compared to same age control cells. Such alterations in energy metabolism in fetal IUGR cardiomyocytes are consistent with an early metabolic shift to favor lipids as the main energy source. This shift normally occurs in the neonatal life to adapt to an extra-uterine environment. Therefore, our findings suggest that adaptation to an altered placental perfusion results in an early cardiac metabolic remodeling that might predispose individuals to metabolic disorders later in their life. Sources of Research Support: Fond de Recherche Québécois en Santé (FRQS) and Canadian Institutes of Health Research (CIHR).

Insufficient development of the uteroplacental circulation may contribute to the development of intrauterine growth restriction (IUGR). We developed a rat model of IUGR by administering a low-Na+ diet. This diet reduces maternal blood volume expansion and uteroplacental perfusion. We hypothesized that an impaired endothelial function in radial arteries decreases vasorelaxation and lowers placental perfusion in this IUGR model. The objective was to assess radial uterine artery responses to vasoactive agents in the IUGR model versus controls. The vasoactive agents included phenylephrine and carbachol, use of a pressurized artery myograph, in the absence or presence of inhibitors of nitric oxide (NO) synthase [ N-nitro-l-arginine methyl ester (l-NAME)], cyclooxygenase (Ibuprofen), and endothelium-dependent hyperpolarization {apamin/1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole}, allowing better characterization of the mechanism implicated in endothelium-dependent relaxation. The results show that 1) the diameter of uterine radial arteries was significantly decreased in the IUGR group; 2) sensitivity to phenylephrine was reduced in IUGR arteries, which could be returned to control group values by inhibition of NO production; 3) the relaxation response to carbachol was increased in IUGR rats, principally mediated by endothelium-dependent hyperpolarization in both groups; 4) NO synthase inhibition by l-NAME decreased the maximum relaxation to carbachol only in the IUGR group; and 5) relaxation response to NO donors is increased in IUGR compared with control radial arteries. Contrary to the hypothesis, results in the IUGR model indicate that the NO pathway is activated in radial uterine arteries, most likely in compensation for the reduction in blood uteroplacental perfusion. NEW & NOTEWORTHY In contrast to genetic or surgical models of intrauterine growth restriction, the diet-induced model of reduced maternal volume expansion shows the nitric oxide pathway to be activated in the uterine artery, possibly from increased shear stress and/or placental factors.

Lower maternal plasma volume expansion was found in idiopathic intrauterine growth restriction (IUGR) but the link remains to be elucidated. An animal model of IUGR was developed by giving a low-sodium diet to rats over the last week of gestation. This treatment prevents full expansion of maternal circulating volume and the increase in uterine artery diameter, leading to reduced placental weight compared to normal gestation. We aimed to verify whether this is associated with reduced remodeling of uteroplacental circulation and placental hypoxia. Dams were divided into two groups: IUGR group and normal-fed controls. Blood velocity waveforms in the main uterine artery were obtained by Doppler sonography on days 14, 18 and 21 of pregnancy. On day 22 (term = 23 days), rats were sacrificed and placentas and uterine radial arteries were collected. Diameter and myogenic response of uterine arteries supplying placentas were determined while expression of hypoxia-modulated genes (HIF-1α, VEGFA and VEGFR2), apoptotic enzyme (Caspase -3 and -9) and glycogen cells clusters were measured in control and IUGR term-placentas. In the IUGR group, impaired blood velocity in the main uterine artery along with increased resistance index was observed without alteration in umbilical artery blood velocity. Radial uterine artery diameter was reduced while myogenic response was increased. IUGR placentas displayed increased expression of hypoxia markers without change in the caspases and increased glycogen cells in the junctional zone. The present data suggest that reduced placental and fetal growth in our IUGR model may be mediated, in part, through reduced maternal uteroplacental blood flow and increased placental hypoxia.

Placenta being devoid of autonomic innervation, ombilical-placental vascular tone should be regulated by tissular and humoral factors. Among the numerous stimuli able to challenge the placental circulation, we propose that prostanoids could be responsible for placental vascular tone. Consequently, we measured the vasomotor responses of U-46619, a mimetic of the Thromboxane A2 (TXA2), and of the isoprostane 8-iso-Prostaglandin E2 (8-iso-PGE2), in human placental vasculature. Placental cotyledons were set up in a perfusion system. Villous and chorionic arteries were prepared as rings and respectively installed in wire myographs for microvessels and in glass-jacketed tissue baths. The effects of U-46619 and 8-iso-PGE2 were measured in the absence and presence of blockers of TP (TXA2) receptors, SQ29548 and ICI192605, and of EP (PGE2) receptors, AH6809. The influence of nitric oxide (NO) were assessed using L-NAME (10-4 mol/L). U-46619 and 8-iso-PGE2 markedly increased perfusion pressure in cotyledons (> 150 mmHg and > 200 mmHg respectively). In the arteries, U-46619 also increased the tension (villous arteries > 1 mN/mm ; chorionic arteries > 4.48 mN/mm). In the cotyledons, as well as in the chorionic arteries, the dose-response curves to U-46619 were competitively shifted to the right by the 3 blockers. However, in the villous arteries, although ICI192605 had similar competitive antagonism as in other preperations, SQ29548 decreased maximum response to U-46619. The dose-response curves to 8-iso-PGE2 were significantly shifted to the right in presence of SQ29548, ICI192605 and AH6809. L-NAME had no effect on dose-response curves to both stimulants. U-46619 and 8-iso-PGE2 had a powerful vasoconstrictor effects in placental vasculature. These effects appeared to be mediated by TP and EP receptors. These results support our postulate that prostanoids have some important regulatory role on placental vascular tone. However, NO did not seem to be involved. (poster)

Lowering and increasing sodium intake in pregnant rats evoke opposite changes in renin-angiotensin-aldosterone system (RAAS) activity and are associated with alterations of blood volume expansion. As augmented uterine blood flow during gestation is linked to increased circulatory volume, we wanted to determine if low- and high-sodium intakes affect the mechanical properties and angiotensin II (AngII) responses of the uterine vasculature. Non-pregnant and pregnant rats received a normal sodium (0.22% Na+) diet. On the 15th day of gestation some animals were moved to a low-sodium (0.03%) diet, whereas others were given NaCl supplementation as beverage (saline, 0.9% or 1.8%) for 7 days. All rats were killed after 7 days of treatment (eve of parturition). Uterine arcuate arteries (>100 microm) were set up in wire myographs under a tension equivalent to 50 mmHg transmural pressure. The pregnancy-associated increase in diameter of the uterine arteries was significantly attenuated on the low-sodium diet and 1.8% NaCl supplementation. The arcuate arteries of non-pregnant rats on the low-sodium diet showed markedly increased responses to AngII and phenylephrine (Phe). Pregnancy also resulted in heightened responses to AngII and Phe that were significantly reduced for the former agent in rats on the low-sodium diet. Sodium supplementation of non-pregnant rats did not affect the reactivity of the uterine arteries to AngII, but significantly reduced the effect of Phe (1 micromol/l). High salt also significantly diminished the elevated responses to AngII in the arteries of pregnant animals. It was observed that altered sodium intake affects the mechanical and reactive properties of the uterine arcuate arteries more importantly in pregnant than in non-pregnant rats. Low-salt intake similarly affected the reactivity of the uterine arcuate arteries to AngII and Phe, whereas high-salt intake more specifically affected AngII responses. These results showed that perturbations of sodium intake have major impacts on the structure and functions of the uterine arterial circulation, indicating RAAS involvement in uterine vascular remodeling and function during gestation.

We previously reported that sodium restriction during pregnancy reduces plasma volume expansion and promotes intra‐uterine growth restriction (IUGR) in rats while it activates the renin–angiotensin–aldosterone system (RAAS). In the present study, we proceeded to determine whether expression of the two angiotensin II (ANGII) receptor subtypes (AT 1 and AT 2 ) change in relation to maternal water–electrolyte homeostasis and fetal growth. To this end, pregnant (gestation day 15) and non‐pregnant Sprague‐Dawley rats were randomly assigned to two groups fed either normal, or Na + ‐restricted diets for 7 days. At the end of the treatment period, plasma aldosterone and renin activity as well as plasma and urine electrolytes were measured. Determinations for AT 1 and AT 2 mRNA and protein were made by RNase protection assay and photoaffinity labelling, respectively, using a number of tissues implicated in volume regulation and fetal growth. In non‐pregnant rats, Na + restriction decreases Na + excretion without altering plasma volume, plasma Na + concentration or the expression of AT 1 and AT 2 mRNA or protein in the tissues examined. In normally fed pregnant rats when compared to non‐pregnant controls, AT 1 mRNA increases in the hypothalamus as well as pituitary and declines in uterine arteries, while AT 1 protein decreases in the kidney and AT 2 mRNA declines in the adrenal cortex. In pregnant rats, Na + restriction induces a decrease in plasma Na + , an increase in plasma urea, as well as a decline in renal urea and creatinine clearance rates. Protein levels for both AT 1 and AT 2 in the pituitary and AT 2 mRNA in the adrenal cortex are lower in the Na + ‐restricted pregnant group when compared to normally fed pregnant animals. Na + restriction also induces a decrease in AT 1 protein in the placenta. In conclusion, these results suggest that pregnancy may increase sensitivity to Na + depletion by the tissue‐specific modulation of ANGII receptors. Finally, these receptors may be implicated in the IUGR response to low Na + .

Since vascular complications often accompany diabetes, we examined the influence of the endothelial lining on vascular reactivity in Psammomys obesus, a desert gerbil that acquires insulin resistance and diabetes when exposed to a laboratory diet. Vasoconstriction to phenylephrine and depolarizing KCl, as well as carbachol endothelium-dependent relaxation, were assessed in rings of thoracic aortae obtained from three groups: (i) group A, normoglycemic-normoinsulinemic; (ii) group B, normoglycemic-hyperinsulinemic, and (iii) group C, hyperglycemic-hyperinsulinemic animals. As expected, marked hypertriglyceridemia and hypercholesterolemia characterized groups B and C, which developed enhanced contractile responsiveness to phenylephrine and KCl compared with controls (group A). Furthermore, both experimental groups displayed a significant decrease in endothelium-dependent relaxation to carbachol. Altered lipid profiles are considered to play some role in the observed modification of aortic reactivity. Overall, our data indicate that vascular contractile responsiveness is enhanced early in the development of insulin resistance and diabetes in the female P. obesus.

During pregnancy, uterine circulation undergoes hypertrophy and hyperplasia. We investigated the effects of angiotensin (Ang) II receptor subtype (AT(1)/AT(2)) blockade on increased responses to the peptide during reversible remodeling of the uterine vasculature in pregnant and postpartum rats. Uterine arcuate arteries were set up in wire myographs for microvessel and submitted to a tension equivalent to 50 mm Hg transmural pressure. Cumulative concentration-response curves to Ang II were measured in the absence and presence of losartan on the same vascular segment. A similar protocol was repeated in the presence of PD 123,319, an AT(2) receptor blocker, again in the absence and presence of losartan. Responses to Ang II on the arcuate artery increased markedly during pregnancy and returned to the prepregnant level within 12 days postpartum. Losartan (10(-7) mol/L) produced a parallel right shift of the concentration-response curve to Ang II in all groups of tissues, but potency of the AT(1) receptor blocker was reduced at the end of pregnancy and in the early postpartum period. PD 123,319 (10(-7) mol/L) significantly increased maximum response to Ang II in arterial segments of the nonpregnant, term-pregnant, and 5 days postpartum rats. AT(1) receptor expression was decreased in arcuate arteries of term-pregnant rats. These results show that contractile responses to Ang II on the uterine arcuate artery of the rat are mediated by the AT(1) receptor and that blockade of AT(2) receptors potentiated responses to the peptide. They also indicate that, in uterine vessels, AT(2) receptor stimulation interferes with Ang II responses, but this effect is decreased in uterine arcuate arteries in the peripartum period.