Model steatogenic compounds (amiodarone, valproic acid, and tetracycline) alter lipid metabolism by different mechanisms in mouse liver slices.
Although drug induced steatosis represents a mild type of hepatotoxicity it can progress into more severe non-alcoholic steatohepatitis. Current models used for safety assessment in drug development and chemical risk assessment do not accurately predict steatosis in humans. Therefore, new models need to be developed to screen compounds for steatogenic properties. We have studied the usefulness of mouse precision-cut liver slices (PCLS) as an alternative to animal testing to gain more insight into the mechanisms involved in the steatogenesis. To this end, PCLS were incubated 24 h with the model steatogenic compounds: amiodarone (AMI), valproic acid (VA), and tetracycline (TET). Transcriptome analysis using DNA microarrays was used to identify genes and processes affected by these compounds. AMI and VA upregulated lipid metabolism, whereas processes associated with extracellular matrix remodelling and inflammation were downregulated. TET downregulated mitochondrial functions, lipid metabolism, and fibrosis. Furthermore, on the basis of the transcriptomics data it was hypothesized that all three compounds affect peroxisome proliferator activated-receptor (PPAR) signaling. Application of PPAR reporter assays classified AMI and VA as PPARγ and triple PPARα/(β/δ)/γ agonist, respectively, whereas TET had no effect on any of the PPARs. Some of the differentially expressed genes were considered as potential candidate biomarkers to identify PPAR agonists (i.e. AMI and VA) or compounds impairing mitochondrial functions (i.e. TET). Finally, comparison of our findings with publicly available transcriptomics data showed that a number of processes altered in the mouse PCLS was also affected in mouse livers and human primary hepatocytes exposed to known PPAR agonists. Thus mouse PCLS are a valuable model to identify early mechanisms of action of compounds altering lipid metabolism.
Cluster of Bioassays and Toxicology, RIKILT - Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
Ewa
E
Szalowska
BDS BioDetection Systems, Amsterdam, The Netherlands.
Bart
B
van der Burg
BDS BioDetection Systems, Amsterdam, The Netherlands.
Hai-Yen
HY
Man
Cluster of Bioassays and Toxicology, RIKILT - Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
Peter J M
PJ
Hendriksen
Cluster of Bioassays and Toxicology, RIKILT - Institute of Food Safety, Wageningen University and Research Centre, Wageningen, The Netherlands.
Ad A C M
AA
Peijnenburg
D000638:Amiodarone / Q000494:pharmacology*; D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005234:Fatty Liver / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D047493:PPAR alpha / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D047495:PPAR gamma / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D048068:PPAR-beta / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology*; D046509:Tissue Culture Techniques; D059467:Transcriptome; D014635:Valproic Acid / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D047493:PPAR alpha; D047495:PPAR gamma; D048068:PPAR-beta; D014635:Valproic Acid; D013752:Tetracycline; D000638:Amiodarone