Ethanol Enhances TGF-p Activity by Recruiting TGF-p Receptors From Intracellular Vesicles/Lip id Rafts/Caveolae to Non-Lipid Raft Microdomains

摘要

Regular consumption of moderate amounts of ethanol has important health benefits on atherosclerotic cardiovascular disease (ASCVD). Overindulgence can cause many diseases, particularly alcoholic liver disease (ALD). The mechanisms by which ethanol causes both beneficial and harmful effects on human health are poorly understood. Here we demonstrate that ethanol enhances TGF-btea-stimulated luciferase activity with a maximum of 0.5-1% (v/v) in MvlLu cells stably expressing a luciferase reporter gene containing Smad2-dependent elements. In MvlLu cells, 0.5% ethanol increases the level of P-Smad2, a canonical TGF-p signaling sensor, by ~2-3-fold. Ethanol (0.5%) increases cell-surface expression of the type II TGF-p receptor (TpR-II) by ~2-3-fold from its intracellular pool, as determined by I125-TGF-p-cross-linking/Western blot analysis. Sucrose density gradient ultracentrifugation and indirect immunofluorescence staining analyses reveal that ethanol (0.5% and 1%) also displaces cell-surface TpR-I and TpR-II from lipid rafts/caveolae and facilitates translocation of these receptors to non-lipid raft microdomains where canonical signaling occurs. These results suggest that ethanol enhances canonical TGF-p signaling by increasing non-lipid raft microdomain localization of the TGF-p receptors. Since TGF-p plays a protective role in ASCVD but can also cause ALD, the TGF-p enhancer activity of ethanol at low and high doses appears to be responsible for both beneficial and harmful effects. Ethanol also disrupts the location of lipid raft/caveolae of other membrane proteins (e.g., neurotransmitter, growth factor/cytokine, and G protein-coupled receptors) which utilize lipid rafts/caveolae as signaling platforms. Displacement of these membrane proteins induced by ethanol may result in a variety of pathologies in nerve, heart and other tissues. J. Cell. Biochem. 117: 860-871, 2016.