title: Nicotine-induced activation of AMP-activated protein kinase inhibits fatty acid synthase in 3T3L1 adipocytes: a role for oxidant stress.
abstract: Recent studies suggest that the AMP-activated protein kinase (AMPK) acts as a major energy sensor and regulator in adipose tissues. The objective of this study was to investigate the role of AMPK in nicotine-induced lipogenesis and lipolysis in 3T3L1 adipocytes. Exposure of 3T3L1 adipocytes to smoking-related concentrations of nicotine increased lipolysis and inhibited fatty acid synthase (FAS) activity in a time- and dose-dependent manner. The effects of nicotine on FAS activity were accompanied by phosphorylation of both AMPK (Thr(172)) and acetyl-CoA carboxylase (ACC; Ser(79)). Nicotine-induced AMPK phosphorylation appeared to be mediated by reactive oxygen species based on the finding that nicotine significantly increased superoxide anions and 3-nitrotyrosine-positive proteins, exogenous peroxynitrite (ONOO(-)) mimicked the effects of nicotine on AMPK, and N-acetylcysteine (NAC) abolished nicotine-enhanced AMPK phosphorylation. Inhibition of AMPK using either pharmacologic (insulin, compound C) or genetic means (overexpression of dominant negative AMPK; AMPK-DN) abolished FAS inhibition induced by nicotine or ONOO(-). Conversely, activation of AMPK by pharmacologic (nicotine, ONOO(-), metformin, and AICAR) or genetic (overexpression of constitutively active AMPK) means inhibited FAS activity. Notably, AMPK activation increased threonine phosphorylation of FAS, and this effect was blocked by adenovirus encoding dominant negative AMPK. Finally, AMPK-dependent FAS phosphorylation was confirmed by (32)P incorporation into FAS in adipocytes. Taken together, our results strongly suggest that nicotine, via ONOO(-) activates AMPK, resulting in enhanced threonine phosphorylation and consequent inhibition of FAS.
authors.affiliation: Vascular Biology Laboratory, Department of Surgery, Graduate School of Medicine, University of Tennessee, Knoxville, Tennessee 37922.
authors.forename: Zhibo
authors.initials: Z
authors.lastname: An
authors.affiliation: Division of Endocrinology and Diabetes, Department of Medicine, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Division of Endocrinology and Diabetes, Department of Medicine, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104.
authors.forename: Ping
authors.initials: P
authors.lastname: Song
authors.affiliation: Division of Endocrinology and Diabetes, Department of Medicine, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104.
authors.forename: Miao
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Vascular Biology Laboratory, Department of Surgery, Graduate School of Medicine, University of Tennessee, Knoxville, Tennessee 37922.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Geng
authors.affiliation: Vascular Biology Laboratory, Department of Surgery, Graduate School of Medicine, University of Tennessee, Knoxville, Tennessee 37922; Division of Endocrinology and Diabetes, Department of Medicine, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104. Electronic address: ming-hui-zou@ouhsc.edu.
authors.forename: Ming-Hui
authors.initials: MH
authors.lastname: Zou
mesh_terms: D041721:3T3-L1 Cells; D055372:AMP-Activated Protein Kinases; D000103:Acetyl-CoA Carboxylase / Q000378:metabolism; D017667:Adipocytes / Q000201:enzymology*; D000818:Animals; D064429:Fatty Acid Synthases / Q000037:antagonists & inhibitors*; D008066:Lipolysis / Q000187:drug effects; D051379:Mice; D009097:Multienzyme Complexes / Q000502:physiology*; D009538:Nicotine / Q000494:pharmacology*; D018384:Oxidative Stress; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D014280:Triglycerides / Q000032:analysis
chemical_list: D009097:Multienzyme Complexes; D014280:Triglycerides; D009538:Nicotine; D064429:Fatty Acid Synthases; D017346:Protein Serine-Threonine Kinases; C491654:Stk11 protein, mouse; D055372:AMP-Activated Protein Kinases; D000103:Acetyl-CoA Carboxylase