Methylation-reprogrammed CHRM3 results in vascular dysfunction in the human umbilical vein following IVF-ET

Abstract

Assisted reproductive technology (ART) has been used globally among infertile couples. However, many epidemiological investigations have indicated that ART is associated with a range of long-term adverse health outcomes in offspring, including cardiovascular disease, obesity and increased plasma lipid levels. Until now, direct evidence has been limited regarding the pathological changes in vascular function in fetuses with ART. In this study, human umbilical cords were collected from healthy normal pregnancies and IVF-ET pregnancies. Vascular functional studies involving acetylcholine (ACh), antagonists of its specific receptors, and L-type calcium channel/PKC-MLC20 phosphorylation pathway specific inhibitors were conducted. Quantitative real-time PCR, Western blotting and methylation analyses were performed on umbilical vein samples. We found that the umbilical vein constriction induced by ACh in the IVF-ET group was significantly attenuated compared with that in the healthy normal pregnancy group, which was not only associated with the hypermethylation of ACh muscarinic receptor subtype 3 (CHRM3) and decreased expression of CHRM3, PKCβ and CaV1.2, but was also related to the reduced phosphorylation of MLC20. The present study revealed that the hypermethylation of CHRM3, leading to a reduction in CHRM3 expression and downregulation of the CaV1.2/PKC-MLC20 phosphorylation pathway, was responsible for the decreased sensitivity to ACh observed in the umbilical vein under IVF-ET conditions. The hypermethylation of CHRM3 caused by IVF-ET might play an important role in altered vasoconstriction and impact cardiovascular systems in the long run.

GDF15 negatively regulates RGS16 involving in abnormal lipid metabolism in IVF-ET male mice offspring

“…It has been reported to increase fasting blood glucose levels [8], blood pressure [8,9], adiposity, triglyceride levels [10], and vascular dysfunction [9,11,12] in IVF children. In our recent studies, we found that umbilical veins of IVF-ET offspring showed abnormal sensitivity to acetylcholine (ACh) or angiotensin II (AII)-mediated contraction, which could be caused by hypo/hypermethylation of the speci c receptors [13,14]. Our ndings strongly supported the idea that IVF-ET could directly lead to changes in fetal vessel functions, which might result in cardiovascular disorders in the long term.…”

In vitro fertilization with frozen embryo transfer increased histamine-mediated contractile sensitivity via PKCβ in human umbilical vein

“…Recent studies reported that angiotensin II-mediated vasoconstrictions were greater in umbilical veins from pregnancy who received IVF-ET when comparing with the control [ 13 ]. But the contractile response to acetylcholine was lower in IVF-ET group [ 12 ]. However, previous vascular functional studies on HUV did not distinguish samples between fresh or frozen ET.…”

“…Due to lack of autonomic innervation, vessel tone of human umbilical vessels is mainly regulated by vasoactive substances released locally or carried by the blood stream, such as histamine [ 10 , 11 ]. Recent studies reported that IVF-ET could alter the constriction caused by angiotensin II and acetylcholine in the HUVs via abnormal DNA methylation [ 12 , 13 ]. However, there have been very limited data on the effects of frozen ET process on vascular functions in human.…”

“…Activated PKC can phosphorylate ion channels so as to regulate vessel tone [ 29 ]. Pan-inhibitor for PKC, GF109203X could partially decrease acetylcholine-mediated constriction in HUV from non-IVF pregnancies, demonstrating its vital role in regulating umbilical vessel tone [ 12 ]. The present study determined vascular tension using the agonist and inhibitor for PKC following frozen ET treatments.…”

Associations of genetic variations in the M3 receptor with salt sensitivity, longitudinal changes in blood pressure and the incidence of hypertension in Chinese adults

“…M3R is widely distributed in the human heart, has an inotropic effect, and plays vital roles in various cardiac functions, including the regulation of heart rate, mitigation of myocardial ischemic injury, facilitation of cell‐to‐cell communication, and prevention of atrial fibrillation 15,16 . In addition, M3R is expressed in the endothelial cell layer and smooth muscle cells of the vascular system, where it mediates vasodilation and vasoconstriction 17,18 . Moreover, M3R is involved in the regulation of BP.…”