Multiomics reveal non-alcoholic fatty liver disease in rats following chronic exposure to an ultra-low dose of Roundup herbicide

The impairment of liver function by low environmentally relevant doses of glyphosate-based herbicides
(GBH) is still a debatable and unresolved matter. Previously we have shown that rats administered for
2 years with 0.1 ppb (50 ng/L glyphosate equivalent dilution; 4 ng/kg body weight/day daily intake) of a
Roundup GBH formulation showed signs of enhanced liver injury as indicated by anatomorphological,
blood/urine biochemical changes and transcriptome profiling. Here we present a multiomic study
combining metabolome and proteome liver analyses to obtain further insight into the Roundup-
induced pathology. Proteins significantly disturbed (214 out of 1906 detected, q < 0.05) were
involved in organonitrogen metabolism and fatty acid β-oxidation. Proteome disturbances reflected
peroxisomal proliferation, steatosis and necrosis. The metabolome analysis (55 metabolites altered out
of 673 detected, p < 0.05) confirmed lipotoxic conditions and oxidative stress by showing an activation
of glutathione and ascorbate free radical scavenger systems. Additionally, we found metabolite
alterations associated with hallmarks of hepatotoxicity such as γ-glutamyl dipeptides, acylcarnitines,
and proline derivatives. Overall, metabolome and proteome disturbances showed a substantial overlap
with biomarkers of non-alcoholic fatty liver disease and its progression to steatohepatosis and thus
confirm liver functional dysfunction resulting from chronic ultra-low dose GBH exposure.


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