Publication: 6-OHDA ile oluşturulan parkinson sıçan modelinde bir kalsiyum kanal blokeri olan isradipin tedavisinin striatumdaki nöronlar üzerindeki etkisinin incelenmesi
Abstract
6-OHDA ile oluşturulan Parkinson sıçan modelinde bir kalsiyum kanal blokeri olan isradipin tedavisinin striatumdaki nöronlar üzerindeki etkisinin incelenmesi Öğrenci Adı: Hatice BORACI, Danışman Adı: Prof.Dr. Ümit S. ŞEHİRLİ Program Adı: Anatomi Amaç: Parkinson hastalığı, başta substantia nigra pars compacta olmak üzere bazal çekirdeklerdeki dopaminerjik nöron kaybıyla karakterize ikinci en sık nörodejeneratif hastalıktır. Etiyolojsinde mitokondrial disfonksiyon, oksidatif stres ve intraselüler kalsiyum artışı gibi faktörler öne çıkmaktadır. Kalbindin, kalsiyum kanallarını regüle etmektedir. Bu çalışma, lezyon öncesi ve sonrası kan ve beyindeki kalbindin düzeyi değişimi ve kalsiyum kanal blokeri olan isradipinin, kalbindin üzerine etkisini gözlemlemek amacıyla planlanmıştır. Gereç ve yöntem: 6-OHDA, intrakranial stereotaksik yöntemle enjekte edilmiştir. Deneyin birinci kısmında, süreç takibi yapılarak kalbindinin kandaki düzeylerinin değişimi lezyon öncesi, sonrası, 2.,5. ve 10.haftalarda kontrol (sağlıklı) ve lezyonlu gruplarda dekapite edilen beyinlerde, ELİSA yöntemiyle ölçülmüştür. 5. ve 10.haftada ise beyinde kalbindin düzeyi western blot yöntemiyle ölçülmüştür. İkinci kısımda ise, lezyonlu ve kontrol grubuna isradipin (2mg/ kg) subkutenöz olarak 4 hafta süresince verilmiştir. 21.günde apomorfin (0.05mg/ kg) rotasyon testi uygulanmıştır. Beyin kesitlerine sırasıyla tirozin hidroksilaz ve kalbindin immunohistokimya prosedürleri uygulanmıştır. Veriler, two-way ANOVA post-hoc Tukey test in Graph-Pad PRISM 6.0 (GraphPad Software Inc., San Diego, CA, USA) kullanılarak analiz edilmiştir. Bulgular: 5.haftadaki beyinlerde kalbindin düzeyinde kontrol grubuna kıyasla anlamlı bir azalma gözlemlenmiştir (p<0.05). Parkinsonlu isradipin grubuna ait sıçanlarda kalbindin kan düzeyinde Parkinsonlu olup herhangi bir enjeksiyon yapılmayan gruba kıyasla anlamlı bir artış gözlemlenmiştir (p=0.04), rotasyon sayısı anlamlı azalmıştır (p=0,02). 6-OHDA grubunda lezyon öncesi 2.saat ve 2.hafta arasındaki kan kalbindin düzeyinde ise anlamlı bir azalma gözlemlenmiştir (p=0.028) Sonuç: Kalbindinin lezyon sonrası kanda ve beyinde azalışı klinik çalışmalara teşhis için ışık tutmaktadır. Ayrıca, isradipinin ise kanda kalbindin düzeyine etki edebildiğini yaptığımız bu çalışma ile ortaya koymaktayız.
The effect of a calcium channel blocker, isradipine, on calbindin blood levels and calbindin positive neurons in the striatum of Parkinsonian rats lesioned with 6-OHDA Student Name: Hatice BORACI, Name of Supervisor: Prof. Dr. Ümit S. ŞEHİRLİ Program Name: Anatomy Department Objective: Parkinson’s disease (PD) is characterized by loss of the dopaminergic neurons in the substantia nigra pars compacta. In the pathology, imbalance of the intracellular calcium level may appear as a consequence of abnormality related to calcium-binding proteins like calbindin. We examined the time-dependent change of calbindin and the effect of a calcium channel blocker, isradipine, on the blood level of calbindin and neurons. Material and method: 6-OHDA was injected into the rat brains. For the first part of experiment, control and lesioned groups were inspected during the 2nd, 5th, and 10th weeks. Calbindin blood level was assessed by the ELISA method. The calbindin level in the brain was measured by western blot method. For the second part, the rats were treated with isradipine(2mg/ kg). Brains were immunohistochemically stained by tyrosine-hydroxylase and calbindin. The results were analyzed by Graph-Pad PRISM 6.0. Results: The calbindin level in the brain of 6-OHDA group decreased compared to the control group in the 5th week (p<0.05). The number of rotations was significantly lower (p=0,02) in the treatment group. The calbindin blood level of the treatment group was also (p=0.04). The calbindin blood level of the lesioned groups between the periods (2 hours before and 2 weeks after the lesion) showed a significant decrease (p=0.028). Conclusion: To summarize, this study put forth that the amount of calbindin in the brain and blood calbindin level may show a decrease in the early stage of PD. The mechanism of action of isradipine upsurges the calbindin levels which in turn may improve the pathology of PD.
The effect of a calcium channel blocker, isradipine, on calbindin blood levels and calbindin positive neurons in the striatum of Parkinsonian rats lesioned with 6-OHDA Student Name: Hatice BORACI, Name of Supervisor: Prof. Dr. Ümit S. ŞEHİRLİ Program Name: Anatomy Department Objective: Parkinson’s disease (PD) is characterized by loss of the dopaminergic neurons in the substantia nigra pars compacta. In the pathology, imbalance of the intracellular calcium level may appear as a consequence of abnormality related to calcium-binding proteins like calbindin. We examined the time-dependent change of calbindin and the effect of a calcium channel blocker, isradipine, on the blood level of calbindin and neurons. Material and method: 6-OHDA was injected into the rat brains. For the first part of experiment, control and lesioned groups were inspected during the 2nd, 5th, and 10th weeks. Calbindin blood level was assessed by the ELISA method. The calbindin level in the brain was measured by western blot method. For the second part, the rats were treated with isradipine(2mg/ kg). Brains were immunohistochemically stained by tyrosine-hydroxylase and calbindin. The results were analyzed by Graph-Pad PRISM 6.0. Results: The calbindin level in the brain of 6-OHDA group decreased compared to the control group in the 5th week (p<0.05). The number of rotations was significantly lower (p=0,02) in the treatment group. The calbindin blood level of the treatment group was also (p=0.04). The calbindin blood level of the lesioned groups between the periods (2 hours before and 2 weeks after the lesion) showed a significant decrease (p=0.028). Conclusion: To summarize, this study put forth that the amount of calbindin in the brain and blood calbindin level may show a decrease in the early stage of PD. The mechanism of action of isradipine upsurges the calbindin levels which in turn may improve the pathology of PD.