Microtubules are more stable and more highly acetylated in ethanol-treated hepatic cells

Background/AimsChronic alcohol consumption can lead to serious liver disease. Although the disease progression is clinically well-described, the molecular basis for alcohol-induced hepatotoxicity is not understood.MethodsWe examined hepatocyte-specific, alcohol-induced alterations in microtubule dynamics in WIF-B cells. These cells provide an excellent model for studying alcohol-induced hepatotoxicity; they remain differentiated in culture and metabolize alcohol.ResultsConsistent with reports in other hepatic systems, microtubule polymerization in ethanol-treated WIF-B cells was impaired. However, when viewed by epifluorescence, the microtubules in ethanol-treated cells resembled stable polymers. Antibodies to acetylated α-tubulin confirmed their identity morphologically and revealed biochemically that ethanol-treated cells had approximately three-fold more acetylated α-tubulin than control cells. Livers from ethanol-fed rats also contained increased levels of acetylated α-tubulin. Consistent with increased acetylated α-tubulin levels, microtubules in ethanol-treated WIF-B cells were more stable. Because stability increased with increased time of ethanol exposure or concentration, was prevented by 4-methylpyrazole and was potentiated by cyanamide, we conclude that increased acetylation requires alcohol metabolism and is likely to be mediated by acetaldehyde.ConclusionsEthanol metabolism impairs tubulin polymerization, but once microtubules are formed they are hyperstabilized. These ethanol-induced alterations in microtubule integrity likely have profound effects on hepatocyte function.