Moderate alcohol consumption during pregnancy increases potency of two different drugs (the antifungal fluconazole and the antiepileptic valproate) in inducing craniofacial defects: prediction by the in vitro rat whole embryo culture

Abstract

The prenatal exposure to ethanol (Eth), fluconazole (FLUCO) and sodium valproate (VPA) is related to effects on development, producing characteristic syndromic pictures. Among embryotoxic effects described for the three molecules, the alteration on craniofacial morphogenesis is a common feature in humans and animal models, including rodent embryos developed in vitro. The aim of the present work is to evaluate the developmental effects of low Eth serum concentration (17 mM, corresponding to the legal limit to drive in UK, USA, Canada, and many other countries) in mixture with increasing realistic concentrations of the antifungal drug FLUCO (62.5–500 µM) or with increasing realistic concentrations of the antiepileptic drug VPA (31.25–250 µM). Groups exposed to Eth alone (17–127.5 mM), FLUCO alone (62.5–500 µM) or VPA alone (31.25–750 µM) were also included. The chosen alternative animal model was the post-implantation rat whole embryo culture (WEC). E9.5 embryos were exposed in vitro to the test molecules during the whole test period (48 h, corresponding to the developmental stages characteristics of any vertebrate, for human embryos post-fertilization days 23–31). Data were statistically analyzed and processed for modelling applying the benchmark dose (BMD) and relative potency factor (RPF) approaches. Concentration-related effects on facial outcomes were observed in all experimental groups, with a significant enhancement in the groups co-exposed with Eth in comparison to the single exposures. Data obtained by the present work suggest an additional alert for the assumption of even low levels of alcohol in pregnant women during FLUCO or VPA therapy.

Valproic Acid Syndrome: A Mini Review

“…Studies children exposed to valproate during the gestational period showed that the absolute risk of autism spectrum disorder (ASD) was significant (4.15%) [40], and experiments with murine models proved that the intrauterine exposure corroborated the development of problems similar to ASD [25,36,41,42,43], as the drug compromises morphologically the neuronal networks, the molecular markers, and reduces the number of Nissl neurons [44]; it generates musculoskeletal defects associated with a hyper acetylation of histones in somites [45] via the inhibition of histone deacetylases with consequent abnormalities in gill arches due to problems in the gene expression regulated by retinoic acid [46,47,48].…”

“…Among the orofacial abnormalities, there may be a cleft lip and/or palate [48], epicanthic folds, a decreased nose size with anteverted nostrils, an elongated and flat lip filter, an altered cephalic index, craniosynostosis [42], a frontal protrusion, a wide nasal bridge, a bitemporal narrowing [50].…”