3A,B). In order to determine whether MitoQ had an effect on the ethanol metabolizing enzymes, we measured protein expression levels of CYP2E1 and ALDH2 in liver homogenates (Supporting Fig. 2A,B). Consistent with previous reports, CYP2E1 protein expression was increased in all ethanol-fed animals.12 Importantly, CYP2E1 protein expression was similar in ethanol-fed control animals, indicating similar exposure to ethanol in all treatment groups when compared to treatment with MitoQ. Similarly, ALDH2 protein levels were not affected by ethanol consumption or modified by MitoQ. These data suggest that MitoQ treatment does not affect the key enzymes that are responsible for ethanol metabolism. We next investigated

Selleckchem AZD1152 HQPA whether MitoQ would alter levels of protein kinase (AMPK) and the level of phosphorylation of its downstream target acetyl CoA carboxylase (ACC) because it has been reported that total AMPK levels are decreased upon chronic ethanol consumption when compared

to controls.49 In the current study we observed only modest effects on the AMPK system which only showed significance by MitoQ at 25 mg/kg/day (Supporting Fig. 2C). Furthermore, levels of p-ACC were not different between ethanol-fed animals and their pair-matched controls. MitoQ at 5 mg/kg/day had no effect; however, MitoQ at 25 mg/kg/day had a modest, but significant effect on the p-ACC/total protein ratio in both the control and ethanol-fed animals (Supporting Fig. 2D). Chronic ethanol consumption results in find protocol decreased activity of mitochondrial respiratory chain proteins coded for by mitochondrial DNA.15 Consistent with previous studies, chronic ethanol consumption resulted in decreases in the activities of complex I, III, IV, and V and a small increase in citrate synthase activity in isolated mitochondria but was not changed by MitoQ (Table 2).

As previously shown, chronic ethanol consumption decreased complex I (30 kDa subunit), complex IV (subunits I and IV), and complex III (Rieske FeS) proteins levels, although no effects on complex II or complex III core protein 2 were observed (Supporting 上海皓元医药股份有限公司 Fig. 3).15 Overall, treatment with MitoQ had only a modest effect on complex I, 30 kD subunit and complex IV subunit IV and was only evident at the dose of 5 mg/kg/day MitoQ. Chronic ethanol consumption increased hepatic macro- and microvesicular steatosis compared to the pair-fed controls (Fig. 4). Macrosteatototic vesicles distributed around the pericentral region, in contrast, microvesicular steatosis is predominantly present around the portal tract (zone 1) and to a lesser extent in the pericentral region (Fig. 4A). MitoQ (5 and 25 mg/kg/day) significantly decreased macro- and microsteatosis in ethanol-fed rats. In contrast to macrosteatosis, MitoQ did not demonstrate complete protection of microsteatosis at 25 mg/kg/day (Fig. 4B). MitoQ alone at either dose had no effect on steatosis in the control animals.