N-acetylcysteine inhibits kinase phosphorylation during 3T3-L1 adipocyte differentiation

Redox Rep. 2017 Nov;22(6):265-271. doi: 10.1080/13510002.2016.1223267. Epub 2016 Sep 27.

Abstract

Objectives: Reports investigating the effects of antioxidants on obesity have provided contradictory results. We have previously demonstrated that treatment with the antioxidant N-acetylcysteine (NAC) inhibits cellular triglyceride (Tg) accumulation as well as total cellular monoamine oxidase A (MAOA) expression in 3T3-L1 mature adipocytes (Calzadilla et al., Redox Rep. 2013;210-218). Here we analyzed the role of NAC on adipogenic differentiation pathway.

Methods: Assays were conducted using 3T3-L1 preadipocytes (undifferentiated cells: CC), which are capable of differentiating into mature adipocytes (differentiated cells: DC). We studied the effects of different doses of NAC (0.01 or 1 mM) on DC, to evaluate cellular expression of phospho-JNK½ (pJNK½), phospho-ERK½ (pERK½) and, mitochondrial expression of citrate synthase, fumarate hydratase and MAOA.

Results: Following the differentiation of preadipocytes, an increase in the expression levels of pJNK½ and pERK½ was observed, together with mitotic clonal expansion (MCE). We found that both doses of NAC decreased the expression of pJNK½ and pERK½. Consistent with these results, NAC significantly inhibited MCE and modified the expression of different mitochondrial proteins.

Discussion: Our results suggested that NAC could inhibit Tg and mitochondrial protein expression by preventing both MCE and kinase phosphorylation.

Keywords: 3T3-L1; Antioxidant; MAPK; adipocyte differentiation; kinase phosphorylation; mitocondria; monoamine oxidase A; obesity.

MeSH terms

  • 3T3-L1 Cells
  • Acetylcysteine / pharmacology*
  • Adipocytes / cytology
  • Adipocytes / drug effects*
  • Animals
  • Antioxidants / pharmacology*
  • Cell Differentiation / drug effects
  • Mice
  • Monoamine Oxidase / metabolism
  • Phosphorylation / drug effects

Substances

  • Antioxidants
  • Monoamine Oxidase
  • Acetylcysteine

Grants and funding

This study was supported by the grant UBACYT 20020130200160BA (Universidad de Buenos Aires, Argentina), OAT N°42/2013 (Universidad de Buenos Aires, Argentina) and, the grant SAF-2009-10461 (Ministerio de Ciencia e Innovación Tecnológica, España). Juan Carlos Calvo and Liliana N. Guerra are researchers funded by CONICET (Ministerio de Ciencia y Técnica, Argentina) and Universidad de Buenos Aires.