title: The role of sera from equine grass sickness on apoptosis induction in PC12 Tet-off p53 cell line.
abstract: The pathogenesis of equine grass sickness (EGS) has not fully understood. A better understanding of the exact pathogenesis of diseases can help to make an accurate diagnosis. Previous studies reported some pathological damage of neuronal cells in EGS patients. In this study, primarily cytotoxicity of serum from three clinically EGS-diagnosed horses on PC12 Tet-off (PTO) cells was assessed. Subsequently, the apoptotic tests including cytochrome C release, caspase-3/7 activity measurement and DNA fragmentation assay were conducted to clarify the apoptotic effect of serum from EGS patients. Addition of serum from EGS patients at concentrations higher than 25% on PTO cells resulted in a significant cytotoxicity in Alamar blue reduction assay compared with serum from healthy horses. All three apoptotic endpoints showed that the serum from EGS patients does have capability to induce apoptosis. A remarkable up regulation of cytochrome C release accompanied with concentration- and time-dependent augmentation in caspase-3/7 activity and ultimately DNA fragmentation were observed. Our data suggest that serum from EGS patients might have potentially neurotoxic compounds, which exerts cytotoxic and apoptotic effects on neuronal cells. Moreover, the EGS serum-induced apoptosis attributes to augmentation of cytochrome C release and caspase-3/7 activity.
authors.affiliation: Department of Basic Sciences , Faculty of Veterinary Medicine, Urmia University, Urmia, Iran; ; Department for Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Hassan
authors.initials: H
authors.lastname: Malekinejad
authors.affiliation: Department of Basic Sciences , Faculty of Veterinary Medicine, Urmia University, Urmia, Iran;
authors.forename: Nazli
authors.initials: N
authors.lastname: Alizadeh-Tabrizi
authors.affiliation: Department of Basic Sciences , Faculty of Veterinary Medicine, Urmia University, Urmia, Iran;
authors.forename: Araz
authors.initials: A
authors.lastname: Ostadi
authors.affiliation: Department for Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Johanna
authors.initials: J
authors.lastname: Fink-Gremmels
keywords: Caspase-3/7; Cytochrome C; DNA fragmentation; P53 cells; PC12 Tet-off