Mounting
evidence shows that organophosphate flame retardants (OPFRs),
especially aryl- and halogenated-OPFRs, exert various adverse health
effects on living organisms. This study evaluated the hepatotoxic
effect of trihexyl phosphate (THP) as a long-chain alkyl-OPFR on human
hepatocyte cells (LO2) and mouse hepatocyte cells (AML12) by performing
screening of cytotoxicity in vitro. In combination with transcriptomic
analysis, toxicological mechanisms in vitro were further investigated.
Results showed that THP triggered hepatotoxicity in vitro by altering
four signaling pathways: endoplasmic reticulum (ER) stress, apoptosis,
cell cycle, and the glycolysis signaling pathway. Exposure of LO2
and AML12 liver cells to THP (25 μg/mL) significantly induced
ER stress-mediated apoptosis and cell cycle arrest. Meanwhile, downregulation
of glycolysis caused the blockage of energy metabolism. Furthermore,
the high performance liquid chromatography-quadrupole time-of-flight
mass spectrometry (HPLC-Q-TOF-MS/MS) revealed that much of THP was
absorbed into the cells and displayed stability in the two liver cell
lines. In vivo assays using a mouse model demonstrated that exposure
to THP at 400 mg/kg induced the ballooning degeneration of hepatocytes
in liver tissue, whereas exposure to THP at 800 mg/kg caused acute
liver injury with high alanine aminotransferase levels. This study
provides novel insights into the impact of THP on hepatotoxicity in
vitro and in vivo and uncovers the underlying toxicological mechanisms,
which may serve as a guide for further ecological risk assessment
and reasonable application of alkyl-OPFRs.