N-nitrosocarbofuran induces apoptosis in mouse brain microvascular endothelial cells (bEnd.3).
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.
Jung
Yi-Sook
YS
Department of Physiology, School of Medicine, Ajou University, Suwon, Korea. yisjung@ajou.ac.kr
Kim
Chan-Sik
CS
Park
Hye-Seong
HS
Sohn
Seonghyang
S
Lee
Byung-Hoon
BH
Moon
Chang-Kiu
CK
Lee
Soo Hwan
SH
Baik
Eun Joo
EJ
Moon
Chang-Hyun
CH
D000818:Animals; D017304:Annexin A5 / Q000494:pharmacology; D000873:Anthracenes / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D001921:Brain / Q000098:blood supply*; D002235:Carbofuran / Q000031:analogs & derivatives* / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D005419:Flavonoids / Q000494:pharmacology; D005434:Flow Cytometry; D007093:Imidazoles / Q000494:pharmacology; D051379:Mice; D008833:Microcirculation / Q000187:drug effects; D008854:Microscopy, Electron; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D010575:Pesticides / Q000633:toxicity*; D011725:Pyridines / Q000494:pharmacology; D015398:Signal Transduction; D013997:Time Factors
D017304:Annexin A5; D000873:Anthracenes; D005419:Flavonoids; D007093:Imidazoles; C445260:N-nitrosocarbofuran; D010575:Pesticides; D011725:Pyridines; C432165:pyrazolanthrone; D020928:Mitogen-Activated Protein Kinases; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D002235:Carbofuran