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.
Publication Date: 2015-03-15 PubMed ID: 25992246PubMed Central: PMC4405680
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- Journal Article
Summary
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The research explores the harmful effect of sera or blood serum from horses diagnosed with Equine Grass Sickness (EGS) on PC12 Tet-off (PTO) cells. The findings suggest that EGS patient sera contain potentially neurotoxic compounds that trigger cytotoxicity and cell death (apoptosis) in these cells.
Objective and Methodology
The primary aim of this study is to understand the pathogenesis or the mechanism of disease development of Equine Grass Sickness (EGS) better. The researchers carried out the study by assessing how cytotoxic or harmful the serum from three horses that had been clinically diagnosed with EGS was to PC12 Tet-off (PTO) cells. To elaborate on the effects of this serum on these cells, the team conducted apoptotic tests which include:
- Cytochrome C release
- Measurement of caspase-3/7 activity
- DNA fragmentation assay
Main Findings
After introducing the EGS patient serum to the PTO cells, it was found to cause significant cytotoxicity or cell damage in the cells. This was especially noticeable when the serum from the EGS patients was added to the cells at concentrations higher than 25%. This cytotoxic effect was compared to the impact of serum from healthy horses. Interestingly, through all three apoptotic endpoints, the study discovered that the serum from EGS horses does have the ability to induce cell death or apoptosis.
The release of cytochrome C, a molecule involved in inducing cell death, was notably increased. This was accompanied by a concentration- and time-dependent increase in caspase-3/7 activity, which is directly linked to apoptosis. Finally, DNA fragmentation, also a sign of apoptosis, was observed.
Conclusion
The study’s results indicate that neurotoxic compounds in the EGS patient’s serum cause harm and induce apoptosis in PTO cells. Furthermore, this EGS serum-induced cell death is attributed to the increase in cytochrome C release and caspase-3/7 activity. The findings, therefore, suggest a potential link between these neurotoxic compounds and the onset or progression of Equine Grass Sickness. These insights would be useful in formulating diagnostic methods and treatments for this disease in the future.
Cite This Article
APA
Malekinejad H, Alizadeh-Tabrizi N, Ostadi A, Fink-Gremmels J.
(2015).
The role of sera from equine grass sickness on apoptosis induction in PC12 Tet-off p53 cell line.
Vet Res Forum, 6(1), 9-15.
Publication
Researcher Affiliations
- 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.
- Department of Basic Sciences , Faculty of Veterinary Medicine, Urmia University, Urmia, Iran;
- 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.
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