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Published on: May 4, 2021
N-acetylcysteine inhibits kinase phosphorylation during 3T3-L1 adipocyte differentiation
Daniela Soto1, María Gomez-Serrano2, Azul Pieralisi1
1a Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales , Universidad de Buenos Aires - IQUIBICEN, CONICET , Buenos Aires , Argentina.
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.
Insights
N-acetylcysteine (NAC) inhibits adipocyte differentiation by reducing triglyceride accumulation and mitochondrial protein expression. This antioxidant prevents mitotic clonal expansion and key kinase phosphorylation, offering potential therapeutic insights for obesity.
Area of Science:
- Cell Biology
- Biochemistry
- Metabolism
Background:
- Antioxidant effects on obesity are debated.
- N-acetylcysteine (NAC) previously inhibited triglyceride accumulation and MAOA expression in adipocytes.
- The role of NAC in adipogenic differentiation requires further investigation.
Purpose of the Study:
- To analyze the role of NAC in the adipogenic differentiation pathway.
- To evaluate the effects of NAC on key signaling molecules and mitochondrial proteins during adipocyte differentiation.
Main Methods:
- 3T3-L1 preadipocytes were differentiated into mature adipocytes.
- Differentiated cells were treated with varying doses of NAC (0.01 or 1 mM).
- Expression of phospho-JNK½, phospho-ERK½, and mitochondrial proteins (citrate synthase, fumarate hydratase, MAOA) was assessed.
Main Results:
- Differentiation increased phospho-JNK½ and phospho-ERK½ expression and mitotic clonal expansion (MCE).
- NAC treatment decreased phospho-JNK½ and phospho-ERK½ levels.
- NAC significantly inhibited MCE and altered mitochondrial protein expression.
Conclusions:
- NAC inhibits adipogenesis by preventing MCE and kinase phosphorylation.
- NAC reduces triglyceride and mitochondrial protein accumulation.
- These findings suggest NAC's potential in managing obesity-related metabolic dysfunction.
