engleski

IDENTIFICATION OF POTENTIAL DRUG-TARGETS FOR DRUG INDUCED FATTY LIVER DISEASE

Background: Drug-induced fatty liver disease (DIFLD) is strongly associated with duration and dose of medication. Number of drugs have been implicated in the development of chronic DILI, however, the exact type of steatosis and molecular mechanisms of DIFLD are yet not fully enlightened. Therefore, the aim of our study was to define the type of steatosis for tested drugs, to assess their impact on hepatocyte survival and to evaluate molecular mechanisms involved in development of DIFLD in vitro. Methods: HuH7, a well differentiated hepatoma cell line, was used as a model of DIFLD. Cells were exposed to different concentrations of amiodarone (Amiodarone hydrochloride 98%, Sigma Aldrich) and tamoxifen (Tamoxifen citrate 98%, Acros Organics). After 48 hours, the extent of steatosis, apoptosis and lipogenic pathway’ genes expression was evaluated by Oil-Red-O staining, Trypan blue staining, MTT assay and PCR, respectively. Results: Massive increase in the number and size of lipid droplets into the cytosol of drugs-treated cells compared to controls was demonstrated, with peripheral accumulation of droplets on the inner side of cell membrane without displacement of the nucleus and decrease in cell number proportional to concentration of drugs was shown by Oil-Red-O staining and Trypan blue staining, respectively. Tamoxifen showed greater potential of causing the cell death. Additionally, mRNA expression of SREBP1 was upregulated in cells treated with tested drugs compared to controls, while results considering PPARy were not as consistent. Conclusions: Hepatotoxic drugs induced upregulation of lipogenic genes, thus ehnancing triglyceride accumulation and steatogenesis in Huh7 cell cultures. This steatogenic effect was demonstrated by Oil Red-O–staining of the lipid vesicles. Both drugs share a common hepatotoxicity mechanism involving mitochondrial ATP formation.

amiodarone ; tamoxifen ; steatosis ; cell culture

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