(62bf) Xenobiotic Metabolism of Myclobutanil in Cultured Hepatocytes

The liver is the primary location of detoxification of xenobiotics. Elucidating the links between central hepatic metabolism and detoxification pathways will provide better understanding of hepatic injury. Flux Balance Analysis (FBA) is a technique that employs extracellular measurements to elucidate intracellular fluxes of a metabolic network using an optimization framework. FBA in conjunction with pathways analysis can help determine important pathways in the detoxification of the xenobiotics. Conazoles are fungicides that stop the fungal growth by inhibiting ergosterol formation. Various gene expression studies were not able to identify a unique set of toxicological responses that could differentiate the conazoles. The aim of this study is to test the effect of various doses of myclobutanil (a conazole possessing a 1,2,4-triazole ring) on primary rat hepatocytes over a three day culture period. In three six-well plates, hepatocytes were cultured and tested using 4 concentrations ranging from 1.2 mM to 0.15 mM with controls. Initial analysis indicated that higher concentrations (>0.6 mM) of myclobutanil result in cell death over the three day period. Urea production, an important marker of hepatic function is also inhibited by myclobutanil. The results from this experiment will be used for future research as guidelines for optimal choice of myclobutanil concentrations without killing most of the hepatocytes.