CeO2 nanoparticles (NPs) have been shown to cause lung
fibrosis, however, the exact underlying molecular mechanisms are poorly understood. In this study, we have conducted a mass spectrometry-based global metabolomic analysis of human bronchial epithelial BEAS-2B cells treated by CeO2 NPs with different aspect ratios and assessed their toxicity on the bronchial epithelial cells by various cell-based functional assays. Although CeO2 NPs at doses ranging from 12.5 μg/mL to 25 μg/mL displayed low cytotoxicity on the bronchial epithelial cells, the metabolomic analysis revealed a number of metabolites in the cellular metabolic pathways of
sphingosine-1-phosphate,
fatty acid oxidation,
inflammation, etc. were significantly altered by CeO2 NPs, especially those with high aspect ratios. More importantly, the robustness of metabolomics findings was further successfully validated in mouse models upon acute and chronic exposures to CeO2 NPs. Mechanistically, CeO2 NPs upregulated
transforming growth factor beta-1 (TGF-β1) levels in BEAS-2B cells in an aspect ratio-dependent manner through enhancing the expression of
early growth response protein 1 (EGR-1). In addition, both in vitro and in vivo studies demonstrated that CeO2 NPs significantly induced the expression of
sphingosine kinase 1 (SHPK1), phosphorylated Smad2/3 and lung
fibrosis markers. Moreover, targeting SPHK1, TGFβ receptor or Smad3 phosphorylation significantly attenuated the
fibrosis-promoting effects of CeO2 NPs, and SPHK1-S1P pathway exerted a greater effect on the TGF-β1-mediated lung
fibrosis compared to the conventional Smad2/3 pathway. Collectively, our studies have identified the metabolomic changes in BEAS-2B cells exposed to CeO2 NPs with different aspect ratios and revealed the subtle changes in metabolic activities that traditional approaches might have missed. More importantly, we have discovered a previously unknown molecular mechanism underlying CeO2 NP-induced lung
fibrosis with different aspect ratios, shedding new insights on the environmental hazard potential of CeO2 NPs.