Response of PXR signaling pathway to simvastatin exposure in mosquitofish (Gambusia affinis) and its histological changes

Abstract

As a widely used lipid lowering agent, simvastatin recently has been frequently detected in aquatic environment and the potential adverse effects from simvastatin exposure to non-target organisms such as fish is worthy of more attention. The aim of this study was to reveal the responses of detoxification system in fish to simvastatin exposure. In this investigation a ubiquitous small freshwater fish, mosquito fish (Gambusia affinis), was employed as test organism, and the transcriptional expression of nucleus transcriptional factor pregnane X receptor (PXR) and its downstream genes, including P-glycoprotein (P-gp), cytochrome 3A (CYP3A), multidrug resistance protein 2 (MRP2), UDP-glucuronosyl transferase (UGT) in mosquito fish were investigated by qRT-PCR methods under the exposure of concentrations of simvastatin (0.5 μg L^-1, 5 μg L^-1, 50 μgL^-1, 500 μg L^-1) for 24 h, 72 h and 168 h. The related enzyme activity (Erythromycin-N-Demethylase, ERND), the protein expression of PXR and the histological changes of liver tissues in fish were also determined via west blotting and transmission electron microscope approaches in the same conditions. Results showed that the mRNA expression of PXR, CYP3A and P-gp showed significantly changes under simvastatin exposure, exhibiting an obvious time/dose-effect relationship with the prolong of exposure time. ERND activity also showed time-effect at 24 h, and western blotting showed PXR protein displaying a dose-effect relationship to some extent. Hepatocyte cellular of mosquito fish exposed to simvastatin (5 μg L^-1, 168 h) exhibited obvious histological changes in form of swelling, incomplete fragmentary structure etc. Overall, simvastatin altered the expression of PXR signaling pathway and subsequently bring about changes in high-levels of mosquito fish.

Keywords: Enzyme activity; Morphological changes; Mosquito fish; PXR; Simvastatin; Transcriptional expression.