Polyphenol-induced AMPK activation and lipid reduction is abolished by
overexpression of a catalytically inactive mutant of SIRT1 (SIRT1H355A) in
HepG2 cells. A, a representative immunoblot of overexpression of
an adenovirus vector encoding a catalytically inactive SIRT1 mutant in HepG2
cells is shown. B, polyphenol-stimulated AMPK phosphorylation is
abolished by the SIRT1H355A mutant under normal glucose conditions. HepG2
cells were infected for 48 h with Ad-GFP or Ad-SIRT1H355A, followed by
treatment with S17834 (10 μm) or resveratrol (50
μm) for 1 h. C and D, polyphenol-stimulated
AMPK signaling is diminished by the SIRT1H355A mutant under high glucose
conditions. HepG2 cells infected with Ad-GFP or Ad-SIRT1H355A were quiesced in
serum-free medium overnight and treated for 24 h without or with 10
μm of resveratrol or S17834 in the presence of high glucose.
E, the lipid-lowering effect of resveratrol is attenuated by the
SIRT1H355A mutant. *, p < 0.05 versus normal glucose in
cells expressing GFP; #, p < 0.05 versus high glucose
alone in cells expressing GFP; ##, p < 0.05 versus
polyphenol treatment in cells expressing GFP (mean ± S.E., n =
4).