Parkinson modeli oluşturulmuş sıçanlarda ozon tedavisinin etkisinin biyokimyasal biyobelirteçler ile araştırılması
Küçük Resim Yok
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/embargoedAccess
Özet
Parkinson hastalığı (PH), beyin ve tüm sinir sisteminde spesifik bölgelerdeki nöronların ilerleyici kaybı ile karakterize olan nörodejeneratif hastalıklar içinde yer alır ve etkileri yıllarca süren, ileri safhalarda hastada şiddetli hafıza kaybı ve ağır fiziksel işlev bozukluklarına yol açan nörodejeneratif bir hastalıktır. Presemptomatik aşama denen, beyinde substantia nigrada dopamin eksikliğine bağlı klinik semptomların ortaya çıkmasından yıllar önce görülen, aslında hastalığın yarattığı ve erken tedavi seçeneği de sunabilecek olan sistemik disfonksiyonlara bağlı pek çok semptomun bu hastalık ile ilişkisinin ispatlanması zordur.
PH, beyinde dopaminin üretildiği yer olan substantia nigranın pars compacta (SNpc) bölümündeki nöronların önemli ölçüde kaybı ile nigrostriatal dopaminerjik yolda ilerleyici dejenerasyonu içermektedir. PH patofizyolojisinde ayırt edici özellik çoğunlukla Lewy cisimcikleri (LC) ve Lewy nöritleri (LN) formunda α-sinüklein agregasyonlarıdır. PH’nin dolaylı yoldan amiloid-β düzeyindeki artış ve oluşan amiloid-β plakları ile ilişkili olduğu ve Tau proteininin PH patolojisinde yüksek düzeyde fosforillendiği anlaşılmıştır. Ayrıca PH patolojisinde nöronların tirozin hidroksilaz içeriğinin azaldığı, dokudaki glia hücre sayısının arttığı bilinmektedir. nNOS’un PH patogenezindeki rolü tam olarak anlaşılamamıştır; bazı araştırmalar PH gelişiminde nNOS aktivitesinin arttığı yönünde bulgulara sahipken, bazı araştırmalar PH gelişimi ile nNOS aktivitesi arasında bir ilişki bulamamıştır.
PH’yi tedavi etmek için etkin bir yöntem bulunmamaktadır, uygulamadaki tedavi seçenekleri yan etkileri dolayısıyla hastanın yaşam kalitesine ve genel sağlık durumuna zarar vermektedir. Bu nedenle yeni tedavi arayışlarında yeni yaklaşımlara ihtiyaç duyulmakla beraber α-sinüklein proteini tedavi hedefi olarak karşımıza çıkmaktadır. Ayrıca PH’nin erken tanısını ve takibini sağlayan etkin bir biyobelirteç bulunmamaktadır.
Bu araştırmada PH patofizyolojisinde yer aldığı bilinen bazı proteinlerin düzeylerini hem beyin dokusunda hem de kanda ölçerek bu proteinlerin PH erken tanısında ve izleminde bir biyobelirteç olup olmayacaklarını ve ayrıca ozon uygulamasının PH tedavisinde etkinliğini araştırmayı hedefledik.
Rotenon ile PH modeli oluşturulmuş sıçanlarda modelin oluştuğunun kontrolü davranış testleri (apomorfinle indüklenen dönme testi) yapıldıktan sonra 3 hafta (haftada 3 kez) boyunca intarperitoneal 0,8 mg/kg olacak şekilde ozon tedavisini takiben elde edilen beyin dokuları üzerinde biyokimyasal analizler (α-sinüklein, amiloid-β, Tau, fosforillenmiş Tau) ve histolojik incelemeler (α-sinüklein, glia hücre sayısı, tirozin hidroksilaz (+) hücre sayısı, nNOS (+) hücre sayısı) gerçekleştirildi. Ayrıca kanda α-sinüklein, amiloid-β, Tau, fosforillenmiş Tau düzeyleri ölçüldü.
PH modeli oluşturulan grupta apomorfinle indüklenen dönme testi pozitifti. Bu gruptaki histolojik analizlerde beyin dokusundaki glia hücre sayılarında istatistiksel olarak anlamlı (p<0,05) artış, tirozin hidroksilaz (+) hücre sayısında istatistiksel olarak anlamlı azalış saptandı. Ayrıca biyokimyasal analizlerde beyin dokusundaki α-sinüklein, amiloid-β, Tau, fosforillenmiş Tau düzeyleri ve kan α-sinüklein düzeyi istatistiksel olarak anlamlı (p<0,05) yüksek bulundu. PH modeli oluşturulmuş sıçanlara ozon uygulandıktan sonra yapılan histolojik analizlerde beyin dokusundaki tirozin hidroksilaz (+) hücre sayısında istatistiksel olarak anlamlı (p<0,05) artış, nNOS (+) ve glia hücre sayısında istatistiksel olarak anlamlı (p<0,05) azalış saptandı. Bu gruptaki biyokimyasal analizlerde ise beyin dokusunda ve kanda α-sinüklein düzeyinde istatistiksel anlamlı düşüş saptandı.
Sonuç olarak bu çalışmada deneysel Parkinson modeli oluşturulmuş sıçanlarda ozon uygulanmasının PH patofizyolojisinde yer alan özellikle α-sinüklein düzeylerini düşürmesi, tirozin hidroksilaz düzeylerini artırması, glia düzeylerini düşürmesi bize ozon tedavisinin PH için aranan tedavi seçeneği olabileceğini düşündürtmüştür. Ayrıca deneysel olarak yapılan bu çalışmanın olumlu sonuçlarının, klinik çalışmalara yola açacağını düşünüyoruz. Ayrıca kan α-sinüklein düzeyinin PH modelinde yüksek olarak bulunduğundan dolayı bu protein klinik açıdan PH erken tanısını koymada ve izleminde kullanılabilecek bir biyobelirteç adayı olarak değerlendirilebilir. Bu sonuçları güçlendirmek için hem ozon dozu ve uygulama süresinin optimize edilmesine yönelik hem de klinik çalışmaların yapıldığı ileri araştırmalara ihtiyaç bulunmaktadır.
Parkinson's disease (PD) is one of the neurodegenerative diseases that are characterized by the progressive loss of neurons in specific regions of the brain and the entire nervous system, and is defined as a disease whose effects last for years, leading to severe memory loss and severe physical dysfunction in the patient in advanced stages. It is difficult to prove the relationship of many symptoms related to systemic dysfunctions, which is called the presymptomatic stage, which were seen years before the emergence of clinical symptoms due to dopamine deficiency in the substantia nigra in the brain, actually caused by the disease and could offer us early treatment options. PD involves progressive degeneration in the nigrostriatal dopaminergic pathway with significant loss of neurons in the Substantia Nigra pars compacta (SNpc), where dopamine is produced in the brain. Distinctive features in PD pathophysiology are α-synuclein aggregations in the form of Lewy bodies (LC) and Lewy neurites (LN). It has been understood that PD is indirectly releated with an increase in amyloid- β level and elevated amyloid-β plaques, and Tau protein is highly phosphorylated in PD pathology. In addition, it is known that the tyrosine hydroxylase content of neurons decreases and the number of glial cells in the tissue increases in PD pathology. The role of nNOS in the pathogenesis of PD is not fully understood; in some studies no relationship was found between the development of PD and nNOS activity, but in some others, nNOS activity was risen with the development of PD. There is not an effective treatment strategy for the therapy of PD. Side effects of current therapies are harmful for patients and they reduces the quality of life. It is need to have different approaches for a novel therapy of PD and also α-synuclein protein emerges as a treatment target. In addition, there is no biomarker that provides early diagnosis and follow-up of PD. In this study, we aimed to investigate whether these proteins could be biomarkers in the early diagnosis and follow-up of PD by measuring the levels of some proteins known to be involved in the pathophysiology of PH in both brain tissue and blood, and also to investigate the effectiveness of ozone administration in the treatment of PD. Biochemical analysis (α-synuclein amyloid-β, Tau, phosphorylated Tau) and histological examinations (α-synuclein, glia cell number, tyrosine hydroxylase (+) cell number, nNOS (+) cell number) were performed of the brain tissues obtained from experimental rats with PD that was treated with intarperitoneal 0.8 mg / kg ozone for 3 weeks (3 times a week) after conducting behavioral tests (apomorphine-induced rotation test) to control the formation of the experimental model in the group of PD experimental model that was developed by rotenone. In addition, blood levels of α-synuclein, amyloid-β, Tau, phosphorylated Tau were measured.The apomorphine-induced rotation test was positive in the group of PD experimental model. In histological analyzes in this group, a significant (p <0,05) increase in the number of glia cells in the brain tissue and a significant decrease (p<0,05) in the number of tyrosine hydroxylase (+) cells were found. In addition, high α-synuclein, amyloid-β, Tau, phosphorylate Tau levels in brain tissue and blood α-synuclein were found to be significantly higher (p <0,05) in biochemical analysis. In the histological analysis performed after rats that are experimental model of PD were treated with ozone, tyrosine hydroxylase (+) cell number in brain tissue increased significantly p <0,05), nNOS (+) and glia cell number decreased significantly (p<0,05). Biochemical analyzes showed that there was a significant decrease of α-synuclein levels in brain tissue and blood in this group. In conclusion, ozone treatment to PD experimental model altered the level of proteins in the pathophysiology of PD, decreased α-synuclein levels, increased tyrosine hydroxylase levels, and decreased glia levels this result suggest that of ozone therapy can be an option for PD treatment. We also think that these experimental study results will lead to clinical studies. In addition, since the blood α-synuclein level is high in the PD model, this protein can be clinically considered as a biomarker that can be used in the early diagnosis and monitoring of PD. To strengthen these results, further research is needed for both optimization of ozone dose and duration of the ozone therapy, as well as clinical studies.
Parkinson's disease (PD) is one of the neurodegenerative diseases that are characterized by the progressive loss of neurons in specific regions of the brain and the entire nervous system, and is defined as a disease whose effects last for years, leading to severe memory loss and severe physical dysfunction in the patient in advanced stages. It is difficult to prove the relationship of many symptoms related to systemic dysfunctions, which is called the presymptomatic stage, which were seen years before the emergence of clinical symptoms due to dopamine deficiency in the substantia nigra in the brain, actually caused by the disease and could offer us early treatment options. PD involves progressive degeneration in the nigrostriatal dopaminergic pathway with significant loss of neurons in the Substantia Nigra pars compacta (SNpc), where dopamine is produced in the brain. Distinctive features in PD pathophysiology are α-synuclein aggregations in the form of Lewy bodies (LC) and Lewy neurites (LN). It has been understood that PD is indirectly releated with an increase in amyloid- β level and elevated amyloid-β plaques, and Tau protein is highly phosphorylated in PD pathology. In addition, it is known that the tyrosine hydroxylase content of neurons decreases and the number of glial cells in the tissue increases in PD pathology. The role of nNOS in the pathogenesis of PD is not fully understood; in some studies no relationship was found between the development of PD and nNOS activity, but in some others, nNOS activity was risen with the development of PD. There is not an effective treatment strategy for the therapy of PD. Side effects of current therapies are harmful for patients and they reduces the quality of life. It is need to have different approaches for a novel therapy of PD and also α-synuclein protein emerges as a treatment target. In addition, there is no biomarker that provides early diagnosis and follow-up of PD. In this study, we aimed to investigate whether these proteins could be biomarkers in the early diagnosis and follow-up of PD by measuring the levels of some proteins known to be involved in the pathophysiology of PH in both brain tissue and blood, and also to investigate the effectiveness of ozone administration in the treatment of PD. Biochemical analysis (α-synuclein amyloid-β, Tau, phosphorylated Tau) and histological examinations (α-synuclein, glia cell number, tyrosine hydroxylase (+) cell number, nNOS (+) cell number) were performed of the brain tissues obtained from experimental rats with PD that was treated with intarperitoneal 0.8 mg / kg ozone for 3 weeks (3 times a week) after conducting behavioral tests (apomorphine-induced rotation test) to control the formation of the experimental model in the group of PD experimental model that was developed by rotenone. In addition, blood levels of α-synuclein, amyloid-β, Tau, phosphorylated Tau were measured.The apomorphine-induced rotation test was positive in the group of PD experimental model. In histological analyzes in this group, a significant (p <0,05) increase in the number of glia cells in the brain tissue and a significant decrease (p<0,05) in the number of tyrosine hydroxylase (+) cells were found. In addition, high α-synuclein, amyloid-β, Tau, phosphorylate Tau levels in brain tissue and blood α-synuclein were found to be significantly higher (p <0,05) in biochemical analysis. In the histological analysis performed after rats that are experimental model of PD were treated with ozone, tyrosine hydroxylase (+) cell number in brain tissue increased significantly p <0,05), nNOS (+) and glia cell number decreased significantly (p<0,05). Biochemical analyzes showed that there was a significant decrease of α-synuclein levels in brain tissue and blood in this group. In conclusion, ozone treatment to PD experimental model altered the level of proteins in the pathophysiology of PD, decreased α-synuclein levels, increased tyrosine hydroxylase levels, and decreased glia levels this result suggest that of ozone therapy can be an option for PD treatment. We also think that these experimental study results will lead to clinical studies. In addition, since the blood α-synuclein level is high in the PD model, this protein can be clinically considered as a biomarker that can be used in the early diagnosis and monitoring of PD. To strengthen these results, further research is needed for both optimization of ozone dose and duration of the ozone therapy, as well as clinical studies.
Açıklama
Anahtar Kelimeler
Parkinson Hastalığı, Ozon, Nörodejeneratif Hastalık, Rotenon, Parkinson’s Disease, Ozone, Neurodegenerative Disease, Rotenone
