A link between reduced protein thiol levels and cytotoxicity has been demonstrated in a study conducted with the chemical menadione (Di Monte et al., 1984). In our laboratory, studies with isolated mitochondria showed that DHM, but not MCT, has the ability to oxidize protein thiol groups (Santos et al., 2009). Therefore, to investigate whether this would also happen in hepatocytes, we incubated the isolated hepatocytes with MCT and observed a significant oxidation of –SH groups of proteins at 90 min of incubation. However, when DTT was added, the oxidation of these groups was prevented. Thiol groups, in addition to participating in the Venetoclax supplier antioxidant defense system previously mentioned,

regulate various aspects of cellular Linsitinib supplier function. Among these is the induction of cell death by apoptosis, an activity regulated by the redox state of the thiol groups (Sato et al., 1995). One of the pathways that mediate apoptosis is the mitochondrial pathway (Green and Reed, 1998 and Lemasters et al., 1999), which involves the MPT, a calcium-dependent inner mitochondrial membrane permeabilization. This permeability of the inner membrane is associated with the opening of a pore called the permeability transition

pore. The opening of the pore results in the potential loss of the mitochondrial membrane, swelling of the mitochondria and rupture of the mitochondrial outer membrane (Zoratti and Szabò, 1995 and Halestrap et al., 2002), and it is sufficient to promote the release

of cytochrome c (a component of the electron transport chain that allows the transfer of electrons between complex III and IV) into the cytoplasm of the cell (Kroemer, 1997). Cytochrome c in turn interacts with apoptotic protease activating factors (Apaf), triggering the cascade of activation of pro-caspases by proteolytic cleavage and causing death by apoptosis. By assessing the effects of MCT on the induction of apoptosis with the dye Hoechst 33342 in parallel with monitoring the decrease in cell viability by changes in the pattern of release of the enzyme ALT, we found that MCT is C59 mw able to induce programmed cell death. A possible cause for this observed effect can be found in our previous work with isolated mitochondria (Mingatto et al., 2007). We demonstrated that DHM inhibits NADH-dehydrogenase, causing a significant reduction in the synthesis of ATP, which is a critical event for the development of cell damage by necrosis or apoptosis (Nicotera et al., 1998). In addition, DHM causes the oxidation of thiol groups of proteins from mitochondria, resulting in the release of cytochrome c (Santos et al., 2009), which initiates the cascade of induction of programmed cell death. Accordingly, Copple et al. (2004) showed that MCT kills cultured hepatic parenchymal cells by apoptosis, with activation of caspase 3.