Simvastatin modulates gene expression of key receptors in zebrafish embryos

ABSTRACT

Nuclear receptors (NR) are involved in the regulation of several metabolic processes and it is well known that these constituents may be modulated by different chemicals classes, including pharmaceuticals that may activate or antagonize NR. In mammals, some pharmaceuticals modulate the transcription of pregnane X receptor, Pxr, peroxisome proliferator activated receptor, Ppars, and aryl hydrocarbon receptor, Ahr, affecting mRNA expression of genes belonging to various regulatory pathways, including lipid metabolism and detoxification mechanisms. The aim of this study was to determine the effects of simvastatin (SIM), an anticholesterolemic drug, on selected NR and AhR mRNA transcription levels during zebrafish early development. Embryos were collected at different development stages (0, 2, 6, 14, 24, 48, and 72 hr post fertilization (hpf)) and mRNA of all target NR was detected at all time points. Embryos (1 and 24 hpf) were exposed to different concentrations of SIM (5 or 50 μg/L) in two differing assays with varying exposure times (2 or 80 hr). The transcription levels of ahr2, raraa, rarab, rarga, pparαa, pparβ1, pparγ, pxr, rxraa, rxrab, rxrbb, rxrga, rxrgb, as well as levels of cholesterol (Chol) were measured after exposure. SIM exerted no marked effect on Chol levels, and depending upon exposure duration mRNA levels of NR and AhR either increased or decreased. After 2 hr SIM treatment in 24 hpf embryos, transcription of ppars, pxr, and ahr was up-regulated, while after 80 hr mRNA levels of pxr and ahr were decreased with no marked changes in ppars. Data demonstrate that SIM produced alterations in gene expression of NR which are involved in varying physiological functions and that may disturb regulation of different physiological processes which might impair fish survival and ecosystems regeneration.

Acknowledgments

The authors are thankful to Hugo Santos, Olga Martinez, and Ricardo Lacerda for the technical assistance provided in the maintenance of the aquaria. Authors also thank Ana Capitão for the help with the lipid samples.

Funding

This study was funded by the Portuguese Science and Technology Foundation (FCT) through a fellowship to Virgínia Cunha (SFRH/BD/77931/2011), FCT projects PTDC/MAR/098791/2008, and PSYCHOBASS (PTDC/AAG MAA/2405/2012) and UID/Multi/04423/2013.

Additional information

Funding

This study was funded by the Portuguese Science and Technology Foundation (FCT) through a fellowship to Virgínia Cunha (SFRH/BD/77931/2011), FCT projects PTDC/MAR/098791/2008, and PSYCHOBASS (PTDC/AAG MAA/2405/2012) and UID/Multi/04423/2013.