Expression of testosterone-dependent enzyme, carbonic anhydrase III, and oxidative stress in experimental alcoholic...

We studied the sequential immunohistochemical appearance of androgen-dependent carbonic anhydrase (CA III) during the development of ethanol-induced liver injury using liver samples from castrated and noncastrated male micropigs. In castrated micropigs, the baseline expression of CA III was either low or absent, while distinct positive immunoreactions were found in zone 3 hepatocytes at 5 and 12 months after the initiation of the ethanol diet. The CA III enzyme and protein adducts of lipid peroxidation-derived aldehydic products, malondialdehyde and 4-hydroxynonenal, appeared together in the perivenous region, suggesting that the enzyme functions in an oxidative environment. The positive staining became more abundant and widespread during the progression of alcoholic liver disease. After 12 months, CA III was significantly more abundant in both the ethanol-fed noncastrated and castrated micropigs than in the control animals (P < 0.001, P < 0.05, respectively). CA III content was strikingly high in the ethanol-fed noncastrated animals, consistent with a potential role of androgens in the regulation of ethanol-induced CA III expression. The strongly positive CA III immunoreactions in the ethanol-fed noncastrated micropigs were associated with scant evidence of aldehydic protein adducts and minimal histopathology. Thus, enhanced expression of CA III during ethanol consumption may also account in part for gender differences in the susceptibility for alcohol-induced liver injury.