Unveiling the Effects of Two Polycyclic Aromatic Hydrocarbons and Two Temperatures on the Trout RTL-W1 Cell Line Expression of Detoxification-Related Target Genes
Polycyclic aromatic hydrocarbons (PAHs), prevalent aquatic contaminants, arise from burning fossil fuels, a major source of greenhouse gases driving global warming. PAHs and warmer temperatures individually exert diverse negative effects on aquatic organisms. However, the effects of PAH exposure and...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-06-01
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| Series: | Journal of Xenobiotics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2039-4713/15/3/84 |
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| Summary: | Polycyclic aromatic hydrocarbons (PAHs), prevalent aquatic contaminants, arise from burning fossil fuels, a major source of greenhouse gases driving global warming. PAHs and warmer temperatures individually exert diverse negative effects on aquatic organisms. However, the effects of PAH exposure and/or rising temperature remain largely unknown. Liver in vitro models, like the rainbow trout (<i>Oncorhynchus mykiss</i>) RTL-W1 liver cell line, have been employed to unravel PAH-exposure effects, primarily on cell viability and enzymatic activity. Here, monolayer-cultured (2D) RTL-W1 cells were used to assess the co-exposure effects of temperature (18 and 21 °C) and two PAHs, benzo[a]pyrene (B[a]P) and benzo[k]fluoranthene (B[k]F), at 10 and 100 nM. After a 72 h exposure, the cell density and viability were evaluated using the trypan blue and LDH assays. The mRNA levels of the detoxification-associated genes <i>aryl hydrocarbon receptor</i> (<i>AhR</i>), <i>cytochrome P450 </i>(<i>CYP</i>)<i>1A</i>, <i>CYP3A27</i>, <i>glutathione S-transferase omega 1</i> (<i>GSTO1</i>), <i>uridine diphosphate–glucuronosyltransferase</i> (<i>UGT</i>), <i>catalase</i> (<i>CAT</i>), and <i>multidrug resistance-associated protein 2</i> (<i>MRP2</i>) were measured by RT-qPCR. Temperature influenced cell viability and LDH leakage. Both PAHs reduced the cell density and upregulated the mRNA levels of <i>AhR</i>, <i>CYP1A, CYP3A27</i>, and <i>UGT</i>, while <i>GSTO1</i> and <i>MRP2</i> were only augmented after the higher B[k]F concentration. Temperature influenced <i>CAT</i> and <i>UGT</i> expression. There was no interaction between temperature and the PAHs. Overall, the results show that B[k]F has more effects on detoxification targets than B[a]P, whereas a temperature increase mildly affects gene expression. The RTL-W1 in 2D seems useful for unravelling not only the liver effects of PAH but also the impact of temperature stress. |
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| ISSN: | 2039-4705 2039-4713 |