Carnosic Acid Attenuates an Early Increase in ROS Levels during Adipocyte Differentiation by Suppressing Translation of Nox4 and Inducing Translation of Antioxidant Enzymes

Int J Mol Sci. 2021 Jun 5;22(11):6096. doi: 10.3390/ijms22116096.


The objective of this study was to investigate molecular mechanisms underlying the ability of carnosic acid to attenuate an early increase in reactive oxygen species (ROS) levels during MDI-induced adipocyte differentiation. The levels of superoxide anion and ROS were determined using dihydroethidium (DHE) and 2'-7'-dichlorofluorescin diacetate (DCFH-DA), respectively. Both superoxide anion and ROS levels peaked on the second day of differentiation. They were suppressed by carnosic acid. Carnosic acid attenuates the translation of NADPH (nicotinamide adenine dinucleotide phosphate) oxidase 4 (Nox4), p47phox, and p22phox, and the phosphorylation of nuclear factor-kappa B (NF-κB) and NF-κB inhibitor (IkBa). The translocation of NF-κB into the nucleus was also decreased by carnosic acid. In addition, carnosic acid increased the translation of heme oxygenase-1 (HO-1), γ-glutamylcysteine synthetase (γ-GCSc), and glutathione S-transferase (GST) and both the translation and nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2). Taken together, these results indicate that carnosic acid could down-regulate ROS level in an early stage of MPI-induced adipocyte differentiation by attenuating ROS generation through suppression of NF-κB-mediated translation of Nox4 enzyme and increasing ROS neutralization through induction of Nrf2-mediated translation of phase II antioxidant enzymes such as HO-1, γ-GCS, and GST, leading to its anti-adipogenetic effect.

Keywords: HO-1 translation; IkBα phosphorylation; NF-κB translocation; Nox4 enzyme translation; Nrf2 translocation; ROS generation; ROS neutralization; carnosic acid.

MeSH terms

  • 3T3-L1 Cells
  • ATPases Associated with Diverse Cellular Activities / genetics*
  • Abietanes / pharmacology*
  • Adipocytes / drug effects
  • Adipocytes / metabolism
  • Animals
  • Antioxidants / pharmacology
  • Cell Differentiation / drug effects
  • Cytochrome b Group / genetics
  • DNA Helicases / genetics*
  • Ethidium / analogs & derivatives
  • Ethidium / pharmacology
  • Fluoresceins / pharmacology
  • Glutathione Transferase / genetics
  • Heme Oxygenase-1 / genetics*
  • Membrane Proteins / genetics*
  • Mice
  • NADPH Oxidase 4 / genetics*
  • NADPH Oxidases / genetics
  • NF-KappaB Inhibitor alpha / genetics*
  • Protein Biosynthesis / drug effects
  • Reactive Oxygen Species / metabolism


  • Abietanes
  • Antioxidants
  • Cytochrome b Group
  • Fluoresceins
  • Membrane Proteins
  • Reactive Oxygen Species
  • dihydroethidium
  • NF-KappaB Inhibitor alpha
  • 2',7'-dichlorofluorescein
  • Heme Oxygenase-1
  • Hmox1 protein, mouse
  • NADPH Oxidase 4
  • NADPH Oxidases
  • Cyba protein, mouse
  • Glutathione Transferase
  • ATPases Associated with Diverse Cellular Activities
  • DNA Helicases
  • RUVBL2 protein, mouse
  • Ethidium
  • salvin