Prevention of vascular dysfunction and arterial hypertension in mice generated by assisted reproductive technologies by addition of melatonin to culture media

Abstract

Rexhaj E, Pireva A, Paoloni-Giacobino A, Allemann Y, Cerny D, Dessen P, Sartori C, Scherrer U, Rimoldi SF. Prevention of vascular dysfunction and arterial hypertension in mice generated by assisted reproductive technologies by addition of melatonin to culture media. Am J Physiol Heart Circ Physiol 309: H1151-H1156, 2015. First published August 14, 2015; doi:10.1152/ajpheart.00621.2014.-Assisted reproductive technologies (ART) induce vascular dysfunction in humans and mice. In mice, ART-induced vascular dysfunction is related to epigenetic alteration of the endothelial nitric oxide synthase (eNOS) gene, resulting in decreased vascular eNOS expression and nitrite/nitrate synthesis. Melatonin is involved in epigenetic regulation, and its administration to sterile women improves the success rate of ART. We hypothesized that addition of melatonin to culture media may prevent ART-induced epigenetic and cardiovascular alterations in mice. We, therefore, assessed mesenteric-artery responses to acetylcholine and arterial blood pressure, together with DNA methylation of the eNOS gene promoter in vascular tissue and nitric oxide plasma concentration in 12-wk-old ART mice generated with and without addition of melatonin to culture media and in control mice. As expected, acetylcholine-induced mesenteric-artery dilation was impaired (P ϭ 0.008 vs. control) and mean arterial blood pressure increased (109.5 Ϯ 3.8 vs. 104.0 Ϯ 4.7 mmHg, P ϭ 0.002, ART vs. control) in ART compared with control mice. These alterations were associated with altered DNA methylation of the eNOS gene promoter (P Ͻ 0.001 vs. control) and decreased plasma nitric oxide concentration (10.1 Ϯ 11.1 vs. 29.5 Ϯ 8.0 M) (P Ͻ 0.001 ART vs. control). Addition of melatonin (10 Ϫ6 M) to culture media prevented eNOS dysmethylation (P ϭ 0.005, vs. ART ϩ vehicle), normalized nitric oxide plasma concentration (23.1 Ϯ 14.6 M, P ϭ 0.002 vs. ART ϩ vehicle) and mesentery-artery responsiveness to acetylcholine (P Ͻ 0.008 vs. ART ϩ vehicle), and prevented arterial hypertension (104.6 Ϯ 3.4 mmHg, P Ͻ 0.003 vs. ART ϩ vehicle). These findings provide proof of principle that modification of culture media prevents ART-induced vascular dysfunction. We speculate that this approach will also allow preventing ART-induced premature atherosclerosis in humans. STUDIES IN ANIMALS AND HUMANS demonstrate that adverse events during early life induce vascular dysfunction that may predispose to premature cardiovascular morbidity and mortality (1,2,10,19,24). Assisted reproductive technologies (ART) represent a recent important example of this problem, since ART induces vascular and cardiac dysfunction in children (26,33,34) and causes premature vascular aging and arterial hypertension that are associated with a shortened life span in mice (20). Consistent with observations in humans suggesting that decreased nitric oxide [nitrate/nitrite (NOx)] bioavailability contributes to vascular dysfunction in the offspring of ART (22), data in ART mice indicate that epigenetic alterations of the endotheli...

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“…In a subsequent study, the authors demonstrated that the addition of melatonin in the culture media prevented artery vasodilation, arterial blood pressure, and associated Nos3 promoter methylation effects in IVF mice [54]. While the exact mechanism of melatonin rescue has yet to be elucidated, importantly, this work highlights the importance of embryo culture conditions in the prevention of adverse effects.…”

Effects of perinatal, late foetal, and early embryonic insults on the cardiovascular phenotype in experimental animal models and humans

“…We speculated that addition of melatonin to the culture media may have favourable eff ects on the cardiovascular phenotype in ART mice. We found that addition of melatonin normalized the methylation pattern in the preimplantation embryo and prevented the altered methylation, expression and function of the eNOS gene in vascular tissue and, in turn, prevented mesenteric artery endothelial dysfunction and arterial hypertension in adult ART mice [73] (Figure 7).…”

Melatonin Scavenger Properties against Oxidative and Nitrosative Stress: Impact on <em>In Vitro</em> Embryo Production in Mammals

“…On the other hand, ART can induce vascular dysfunction and arterial hypertension related to epigenetic alterations of the regulation of the eNOS gene; however this can be prevented by addition of melatonin during in vitro culture of embryos which doubled the success rate of IVF, prevented eNOS demethylation and normalized NO plasma concentration [131].…”