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


CONCLUSIONS
Our results suggested that NAC could inhibit Tg and mitochondrial protein expression by preventing both MCE and kinase phosphorylation.
a Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales , Universidad de Buenos Aires - IQUIBICEN, CONICET , Buenos Aires , Argentina.
Daniela
D
Soto
b Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas (CSIC) y Universidad Autónoma de Madrid, (CSIC, UAM) , Madrid , Spain.
María
http://orcid.org/0000-0002-9669-091X
M
Gomez-Serrano
a Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales , Universidad de Buenos Aires - IQUIBICEN, CONICET , Buenos Aires , Argentina.
Azul
A
Pieralisi
c IBYME, CONICET , Buenos Aires , Argentina.
Juan C
http://orcid.org/0000-0001-7632-611X
JC
Calvo
b Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas (CSIC) y Universidad Autónoma de Madrid, (CSIC, UAM) , Madrid , Spain.
Belén
B
Peral
a Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales , Universidad de Buenos Aires - IQUIBICEN, CONICET , Buenos Aires , Argentina.
Liliana N
LN
Guerra
D041721:3T3-L1 Cells; D000111:Acetylcysteine / Q000494:pharmacology*; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D051379:Mice; D008995:Monoamine Oxidase / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects
D000975:Antioxidants; D008995:Monoamine Oxidase; D000111:Acetylcysteine
3T3-L1; Antioxidant; MAPK; adipocyte differentiation; kinase phosphorylation; mitocondria; monoamine oxidase A; obesity
Proc Natl Acad Sci U S A. 2006 Jul 18;103(29):10923-8
16829576
Am J Physiol Endocrinol Metab. 2014 Aug 1;307(3):E262-77
24918202
Mol Cell Biol. 2000 Oct;20(19):7205-13
10982837
Annu Rev Biochem. 1995;64:345-73
7574486
J Mol Cell Biol. 2011 Apr;3(2):123-31
21278199
Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):850-5
12525691
J Mol Med (Berl). 2004 May;82(5):336-44
15007512
Redox Rep. 2013;18(6):210-8
24112955
Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):44-9
12502791
Cancer Res. 2010 Nov 15;70(22):9153-65
20978192
J Biol Chem. 2004 Aug 20;279(34):35914-22
15192111
Nat Rev Mol Cell Biol. 2008 Oct;9(10):747-58
18813292
J Biol Chem. 2009 Apr 17;284(16):10601-9
19237544
EMBO Mol Med. 2013 Aug;5(8):1165-79
23828861
Front Physiol. 2014 Sep 18;5:358
25278906
J Proteomics. 2015 Jun 21;124:79-87
25865306
Anal Biochem. 1976 May 7;72:248-54
942051
J Biol Chem. 2002 Nov 29;277(48):46226-32
12270934
Clin Chem. 2001 Aug;47(8):1467-9
11468240
Nutr Hosp. 2013 May-Jun;28(3):671-5
23848087
Int J Cell Biol. 2012;2012:161837
22888353
FASEB J. 2002 Jul;16(9):1129-31
12039844
EMBO J. 1999 Mar 1;18(5):1321-34
10064598
Circulation. 2005 Nov 22;112(21):3297-305
16286591
Biochimie. 2005 Jan;87(1):51-6
15733737
J Pharmacol Exp Ther. 2015 Nov;355(2):137-44
26311813
Mol Biol Cell. 2011 Jul 1;22(13):2165-74
21562223
Physiol Rev. 2002 Jan;82(1):47-95
11773609
Clin Endocrinol (Oxf). 2014 Oct;81(4):582-7
24628390
Biochem J. 2002 Feb 1;361(Pt 3):621-7
11802792
Growth Horm IGF Res. 2010 Jun;20(3):212-9
20185348
Int J Mol Sci. 2011;12(10):6936-51
22072928
Differentiation. 2009 Feb;77(2):115-20
19281770