Probing the hepatotoxicity mechanisms of novel chlorinated polyfluoroalkyl sulfonates to zebrafish larvae: Implication of structural specificity

Abstract

Hepatotoxicity in zebrafish (Danio rerio) larvae elicited by legacy perfluorooctane sulfonate (PFOS) and its three novel chlorinated alternatives, including chlorinated polyfluorooctane sulfonate (Cl-PFOS) and chlorinated polyfluoroalkyl ether sulfonates (6:2 and 8:2 Cl-PFESA analogs), was evaluated in this study. Upon 7-day separate exposure to the four target compounds at 1 µmol/L, significant hepatic steatosis in exposed larvae was evidenced by pathological micro/macro vacuolation, which was presumably attributed to the excess accumulation of lipid, especially the overloaded triglyceride (TG) level. Disruption on gene transcription was subjected to a structure-dependent manner. In general, PFOS, Cl-PFOS and 6:2 Cl-PFESA of the identical carbon chain length (i.e. C8), despite with different substituents, displayed a similar activation mode and comparable disruptive potency on lipid metabolism responsive genes, which particularly promoted fatty acid synthesis (acetyl-CoA carboxylase, acacb) and β-oxidation (cytochrome P450 enzymes-1A, cyp1a; peroxisomal acyl-CoA oxidase 1, acox1; and acyl-CoA dehy-drogenase, acadm). However, 8:2 Cl-PFESA with a prolonged carbon chain length (i.e. C10), preferentially disturbed fatty acid exportation (apolipoprotein-B100, apob) and triggered a different modulation pattern on fatty acid β-oxidation against the other three compounds. Molecular docking analysis indicated that 8:2 Cl-PFESA exhibited considerably higher peroxisome proliferator-activated receptors (PPARs) antagonism than others, corresponding to its unique suppression effect on fatty acid β-oxidation responsive genes. To our knowledge, this is the first in vivo study reporting hepatotoxicity of Cl-PFOS and Cl-PFESAs to aquatic organisms. Although characterized with different toxic mode-of-action, these novel alternatives can elicit hepatic steatosis as strong as PFOS, stressing the biological risks in view of their global contamination. Keywords: Novel chlorinated polyfluoroalkyl alternatives, Hepatic steatosis, Structure-dependent mode-of-action, Zebrafish larvae