Hepatotoxicity by Acetaminophen and Amiodarone in Zebrafish Embryos

MATERIALS AND METHOD Liver is an essential organ for the detoxification of ingested chemicals and play pivotal role in various biochemical and physiological processes hence liver toxicity is one of the major causes for the withdrawal of newly discovered molecules. Peculiar characteristics viz. high fecundity rate, transparent eggs, less maintenance cost, livelihood in E3 medium without external feeding make Zebrafish (Danio rerio) suitable vertebral model for the hepatotoxicity assay. On 3 day post fertilisation (dpf), embryos were treated with various concentrations of Acetaminophen and Amiodarone for 72 hr. Embryos from both treatment and control groups were observed daily for mortality and other abnormalities. No mortality was observed in Acetaminophen treated groups. At 25 μM, 15 μM and 10 μM Amiodarone concentration, 100%, 78.5% and 64.2% mortality was observed, respectively and captured images of embryos were analysed to evaluate liver necrosis through Image J software on 6 dpf. Significantly decreased mean pixel intensity was observed at 1000 μM, 2000 μM and 5000 μM Acetaminophen concentrations compared to control group. Significantly decreased mean pixel intensity was observed at 5 μM, 7 μM and 10 μM Amiodarone concentrations compared to control group. Based on the results, it is concluded that, NOAEL for Amiodarone and Acetaminophen is 3 μM and 500 μM respectively. Both molecules are potentially hepatotoxic for zebrafish embryos and the results of this study support the use of zebrafish embryos to predict hepatotoxicity in vertebrates as the results obtained are in line with the proven hepatotoxic effects of both the drugs in humans.