Arecoline inhibits the growth of 3T3-L1 preadipocytes via AMP-activated protein kinase and reactive oxygen species pathways

PLoS One. 2018 Jul 16;13(7):e0200508. doi: 10.1371/journal.pone.0200508. eCollection 2018.

Abstract

The present study was designed to investigate the pathways involved in the effect of betel nut arecoline on cell viability in 3T3-L1 preadipocytes. Arecoline, but not arecaidine or guvacine, inhibited preadipocyte viability in a concentration- and time-dependent manner. Arecoline arrested preadipocyte growth in the G2/M phase of the cell cycle; decreased the total levels of cyclin-dependent kinase 1 (CDK1), p21, and p27 proteins; increased p53 and cyclin B1 protein levels; and had no effect on CDK2 protein levels. These results suggested that arecoline selectively affected a particular CDK subfamily. Arecoline inhibited AMP-activated protein kinase (AMPK) activity; conversely, the AMPK activator, AICAR, blocked the arecoline-induced inhibition of cell viability. Pre-treatment with the antioxidant, N-acetylcysteine, prevented the actions of arecoline on cell viability, G2/M growth arrest, reactive oxygen species (ROS) production, and the levels of CDK1, p21, p27, p53, cyclin B1, and phospho-AMPK proteins. These AMPK- and ROS-dependent effects of arecoline on preadipocyte growth may be related to the mechanism underlying the modulatory effect of arecoline on body weight.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells
  • AMP-Activated Protein Kinases / metabolism*
  • Adipocytes / cytology
  • Adipocytes / metabolism*
  • Animals
  • Arecoline / pharmacology*
  • Cell Cycle Proteins / biosynthesis
  • G2 Phase Cell Cycle Checkpoints / drug effects*
  • Gene Expression Regulation / drug effects
  • Mice
  • Reactive Oxygen Species / metabolism*

Substances

  • Cell Cycle Proteins
  • Reactive Oxygen Species
  • Arecoline
  • AMP-Activated Protein Kinases

Grants and funding

This work was supported by the Ministry of Science and Technology, Taiwan [MOST105-2311-B-008-003, MOST-106-2320-B-008-006, and MOST-106-2320-B-008-008-MY3 to Y.-H.K]; the Landseed General Hospital, Taiwan [Y.-H.K]; and the Taoyuan Armed Forces General Hospital [AFTYGH-105-09 to Y.-H.K; and AFTYGH-10611 and AFTYGH-10716 to Y.-W.T]. Landseed General Hospital provided support in the form of salaries for author YCK. The funding bodies had no role in study design; data collection, analysis, and interpretation; and manuscript writing and submission. The specific roles of these authors are articulated in the 'author contributions' section.