Ethanol administration alters the proteolytic activity of hepatic lysosomes.
Protein accumulation in liver cells contributes to alcohol-induced hepatomegaly and is the result of an ethanol-elicited deceleration of protein catabolism (Alcohol Clin Exp Res 13:49, 1989). Because lysosomes are active in the degradation of most hepatic proteins, the present studies were conducted to determine whether ethanol administration altered the proteolytic activities of partially purified hepatic lysosomes. Rats were fed liquid diets containing either ethanol (36% of calories) or isocaloric maltodextrin for periods of 2-34 days. Prior to death, all animals were injected with [3H]leucine to label hepatic proteins. Rats subjected to even brief periods of ethanol feeding (2-8 days) exhibited significant hepatomegaly and hepatic protein accumulation compared with pair-fed control animals. Crude liver homogenates and isolated lysosomal-mitochondrial and cytosolic subfractions were incubated at 37 degrees C, and the acid-soluble radioactivity generated during incubation was measured as an index of proteolysis. At neutral pH, in vitro protein breakdown in incubated liver homogenates and subcellular fractions from control and ethanol-fed rats did not differ significantly. The extent of protein hydrolysis increased when samples were incubated at pH 5.5, which approximates the pH optimum for catalysis by lysosomal acid proteases. Under the latter conditions, partially purified lysosomes from control animals had 2-fold higher levels of proteolysis than corresponding fractions from ethanol-fed rats. The difference in proteolytic capacity appeared to be related to a lower latency and a higher degree of fragility of lysosomes from ethanol-fed rats at the acidic pH.(ABSTRACT TRUNCATED AT 250 WORDS)
Donohue
T M
TM
Liver Study Unit, Department of Veterans Affairs Medical Center, Omaha, NE 68105.
McVicker
D L
DL
Kharbanda
K K
KK
Chaisson
M L
ML
Zetterman
R K
RK
D000135:Acid Phosphatase / Q000378:metabolism; D000428:Alcohol Drinking / Q000009:adverse effects* / Q000473:pathology; D000818:Animals; D002401:Cathepsin B / Q000378:metabolism; D000431:Ethanol / Q000633:toxicity*; D005215:Fasting / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008108:Liver Diseases, Alcoholic / Q000201:enzymology* / Q000473:pathology; D008247:Lysosomes / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D009929:Organ Size / Q000187:drug effects; D010447:Peptide Hydrolases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D001616:beta-Galactosidase / Q000378:metabolism
D011506:Proteins; D000431:Ethanol; D000135:Acid Phosphatase; D001616:beta-Galactosidase; D010447:Peptide Hydrolases; D002401:Cathepsin B