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Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c
Published on: June 29, 2011
N-nitrosocarbofuran induces apoptosis in mouse brain microvascular endothelial cells (bEnd.3)
Yi-Sook Jung1, Chan-Sik Kim, Hye-Seong Park
1Department of Physiology, School of Medicine, Ajou University, Suwon, Korea. yisjung@ajou.ac.kr
Abstract:
In this study, we investigated whether carbofuran, a commonly used carbamate pesticide, and N-nitrosocarbofuran (NOCF), the N-nitroso metabolite of carbofuran, have cytotoxicity in mouse brain microvascular endothelial cells (bEnd.3). Results from the MTT assay in bEnd.3 cells showed that NOCF but not carbofuran caused a remarkable decrease in cell viability. The cell death induced by NOCF appeared to involve apoptosis, based on our results from annexin V staining and electron microscopy. To investigate the mechanism of the NOCF-induced cell death, we examined the effects of selective inhibitors for MAP kinase pathways, PD98059 (for MEK/ERK), SB202190 (for p38 MAP kinase), and SP600125 (for JNK), on the NOCF-induced cell death. The NOCF-induced cell death was significantly reduced by PD98059, but not by SB202190 or SP600125. NOCF increased ERK phosphorylation as early as 15 min after the treatment and this increase was maintained for 2 h. In summary, our results suggest that NOCF can induce apoptotic cell death, at least in part, through the ERK pathway in brain microvascular endothelial cells.
Insights
N-nitrosocarbofuran (NOCF), a metabolite of the pesticide carbofuran, induces apoptosis in mouse brain cells. This cell death is mediated through the ERK pathway, highlighting NOCF
Area of Science:
- Neuroscience
- Toxicology
- Cell Biology
Background:
- Carbamate pesticides like carbofuran are widely used.
- Metabolites of pesticides may pose distinct toxicological risks.
- Brain microvascular endothelial cells are crucial for the blood-brain barrier.
Purpose of the Study:
- To determine the cytotoxicity of carbofuran and its metabolite N-nitrosocarbofuran (NOCF) in mouse brain microvascular endothelial cells (bEnd.3).
- To elucidate the mechanism underlying NOCF-induced cell death, focusing on apoptotic pathways and MAP kinase signaling.
Main Methods:
- MTT assay to assess cell viability.
- Annexin V staining and electron microscopy to detect apoptosis.
- Treatment with MAP kinase pathway inhibitors (PD98059, SB202190, SP600125) to investigate signaling mechanisms.
- Western blot analysis to detect ERK phosphorylation.
Main Results:
- N-nitrosocarbofuran (NOCF) significantly decreased bEnd.3 cell viability, while carbofuran did not.
- NOCF-induced cell death exhibited characteristics of apoptosis.
- NOCF treatment led to increased ERK phosphorylation.
- Inhibition of the MEK/ERK pathway with PD98059 significantly reduced NOCF-induced cell death.
Conclusions:
- N-nitrosocarbofuran (NOCF) is cytotoxic to mouse brain microvascular endothelial cells.
- NOCF induces apoptotic cell death, partly via the ERK signaling pathway.
- These findings highlight the potential neurotoxic effects of carbofuran metabolites.