Evidence of Altered Mitochondrial Protein Expression After Chronic Ethanol Consumption in the Aged Estrogen-Deficient Female Rat Heart.

BACKGROUND: Estrogen loss has been implicated to increase the risk of alcoholic cardiomyopathy in postmenopausal women. The purpose of this study was to identify novel mitochondrial protein targets for the treatment of alcoholic cardiomyopathy in aged women using a state-of-the-art proteomic approach. We hypothesized that chronic ethanol (EtOH) ingestion exacerbates maladaptive mitochondrial protein expression in the aged female heart. METHODS: Adult (3 months) and aged (18 months) F344 ovary-intact or ovariectomized (OVX) rats were randomly assigned an EtOH or control Lieber-DeCarli "all-liquid" diet for 20 weeks. Proteomic analyses were conducted in mitochondria isolated from left ventricles using isobaric tags for relative and absolute quantification (iTRAQ) 8plex labeling and mass spectrometry (n = 3 to 5/group). RESULTS: After EtOH, significant differences (false discovery rate <5%) were observed in electron transport chain components (NADH dehydrogenase [ubiquinone] flavoprotein 2) as well as proteins involved in lipid metabolism (2,4 dienoyl-CoA reductase) and cellular defense (catalase), suggesting a possible link to congestive heart failure. Directional changes in protein levels were confirmed by Western blotting. Additionally, EtOH significantly reduced state 3 mitochondrial respiration in all groups, yet only reduced respiratory control index in the aged OVX rat heart (p < 0.05). CONCLUSIONS: Collectively, the data reveal that EtOH-induced changes in the mitochondrial proteome exacerbate cardiac dysfunction in aged and estrogen-deficient hearts, but not in adult. In conclusion, iTRAQ is a powerful tool for investigating new mitochondrial targets of alcoholic cardiomyopathy.

Comparison of the agar block and Lieber-DeCarli diets to study chronic alcohol consumption in an aging model of Fischer 344 female rats.

INTRODUCTION: Post-menopausal women have a greater risk of developing alcoholic complications compared to age-matched men. Unfortunately, animal models of chronic ethanol consumption with estrogen deficiency are lacking. Here, we characterize the ability of the agar block and Lieber-DeCarli models of chronic ethanol consumption to produce elevated blood alcohol content (BAC) and liver pathology in the F344 postmenopausal animal model of aging. METHODS: Adult (3 mo) and aged (18 mo) F344 ovary-intact or ovariectomized rats were administered ethanol for 14-20 weeks as follows: diet 1, standard chow access, 10% ethanol in drinking water, and 40% ethanol in agar blocks; diet 2, diet 1 plus low phytoestrogen chow (known to affect ethanol metabolism) for the final 4 weeks; diet 3, Lieber-DeCarli all liquid diet with 36% kcal ethanol. Control animals were matched isocalorically with dextrin. RESULTS: For the agar block diet, average BAC was 13±4 mg/dL across groups. BAC was unaffected by reducing dietary phytoestrogen content (12±4 mg/dL), which is known to interfere with ethanol metabolism. Liver pathology was unaffected by the agar block diet. In contrast, the Lieber-DeCarli diet resulted in BAC of 45±5 mg/dL in conjunction with more severe hepatopathology.223 DISCUSSION: We conclude that the Lieber-DeCarli diet produces greater BAC and hepatopathology to study the effects of chronic ethanol administration in the F344 postmenopausal rodent model of aging when compared to an ethanol agar block diet.