Deneysel akut spinal kord hasarında myrıcetın'in nöroprotektif etkilerinin değerlendirilmesi
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Tarih
2023
Yazarlar
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Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi Meram Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç : Bu projemizde antioksidan özelliklere sahip flavionid polifenolik bileşik sınıfının bir üyesi olan Myricetin’i travmatik spinal kord hasarı oluşturulan ratlara uygulayarak Myricetin’in travma sonrası spinal korddaki serbest radikallere bağlı gelişen sekonder hasara yönelik nöroprotektif etkilerini değerlendirdik.
Yöntem : 45 adet wistar albino cinsi rat her grupta 9 adet olmak üzere 5 gruba ayrılarak randomize edildi. Travmatik spinal kord hasarı titanyum çubuklar vasıtası ile ağırlık düşürme tekniği ile gerçekleştirildi.
Grup-1 (sham) ratlara 1.gün laminektomi yapıldı. 7.gün kan ve doku örnekleri alındı.
Grup-2 ratlara 1.gün laminektomi yapıldı. 7 gün boyunca oral gavaj (OG) ile %0,9 luk serum fizyolojik (SF) verildi. 7.gün kan ve doku örnekleri alındı.
Grup-3 ratlara 1.gün laminektomi yapıldı. Titanyum çubuklar ile spinal travma oluşturuldu. 7 gün boyunca oral gavaj (OG) ile %0,9 luk serum fizyolojik (SF) verildi. 7.gün kan ve doku örnekleri alındı.
Grup-4 ratlara 1.gün laminektomi yapıldı. Titanyum çubuklar ile spinal travma oluşturuldu. 7 gün boyunca oral gavaj (OG) ile düşük doz myricetin (5mg/kg/gün) verildi. 7.gün kan ve doku örnekleri alındı.
Grup-5 ratlara 1.gün laminektomi yapıldı. Titanyum çubuklar ile spinal travma oluşturuldu. 7 gün boyunca oral gavaj (OG) ile yüksek doz myricetin (25mg/kg/gün) verildi. 7.gün kan ve doku örnekleri alındı.
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Deneyin 1. ve 7. günlerinde ratların motor muayeneleri için Basso Beattie and Bresnahan (BBB) ve Modifiye Tarlov Skalası testleri uygulandı. Ratlardan elde edilen hasarlı spinal kord dokusu ve kandan interlökin-1beta (IL-1b), interlökin-10 (IL-10), total antioksidan status (TAS), total oksidatif status (TOS), malonildialdehit (MDA) seviyeleri enzyme linked immunosorbent assay (ELISA) yöntemi ile fakültemiz Tıbbi Biyokimya A.D. Araştırma Laboratuvarında çalışılmıştır. Fakültemiz Histoloji ve Embriyoloji A.D. laboratuarında ratlardan elde edilen spinal kord kesitleri hematoksilen eozin (H-E) boyası kullanılarak histoloji laboratuarında ışık mikroskobu altında incelenmiştir. Apoptotik hücreler TUNEL yöntemi ile ApopTag In Situ Apoptosis Detection Kit (Millipore) kullanılarak etiketlenmiştir. TUNEL pozitif hücreler gri maddede 4 mikronluk kesitlerde ışık mikroskobunda 20'lik objektifde sayılarak yüzdeleri alınmıştır. Histopatoloji skoru ve TUNEL pozitif hücre sayısı, TUKEY testi ile tek yönlü varyans analizi (ANOVA) kullanılarak istatistiksel analiz yapılmıştır.
Bulgular : Myricetin, serum ve dokuda dozdan bağımsız olarak IL-10 seviyelerinde artışa sebep oldu. Serum IL-1beta seviyelerinde ilaç dozundan bağımsız düşüş görülürken, doku örneklerinde yüksek doz ilaç grubunda IL-1beta seviyesinde anlamlı düşüş görüldü. Myricetin’in uygulama dozu arttığında daha belirgin antioksidan yanıt oluşturarak TOS seviyesinde düşüşe, TAS seviyesinde yükselişe yol açtığı tespit edildi. Yine Myricetin’in uygulama dozu arttığında oksidatif stresin güçlü belirteçlerinden olan MDA seviyelerinde anlamlı gerilemeye neden olduğu görüldü. Histopatolojik incelemede Myricetin etkisi ile TUNEL pozitif hücre sayısı ve apoptotik indeksin anlamlı olarak azaldığı tespit edildi. Yüksek doz ilaç grubunda diğer deney gruplarına göre en düşük apoptotik indeks tespit edildi. Klinik olarak Basso-Beattie-Bresnahan (BBB) lokomotor derecelendirme ölçeği ve Modifiye Tarloc Skalası, travma sonrası Myricetin’in nörolojik muayenede iyileşme gösterdiği görüldü.
Sonuç : Elde ettiğimiz biyokimyasal veriler, histopatolojik değerlendirme ve nörolojik muayene sonuçlarına göre Myricetin travma sonrası görülen sekonder spinal kord hasarında antiinflamatuar, antiapoptotik, antioksidan ve nöroprotektif etki göstermiştir.
Objective : In this project, we aimed to identify a class of flavonoid polyphenolic compounds with antioxidant properties. By applying Myricetin, which is a member of the family, to rats with traumatic spinal cord injury. Myricetin has a secondary effect on free radicals in the spinalcord after trauma. We evaluated its neuroprotective effects on injury. Method: 45 Wistar albino rats were randomized into 5 groups, 9 in each group was done. Traumatic spinal cord injury, weight reduction model with titanium rodscarried out. Group-1 (sham), rats underwent laminectomy. 7th day spinal cord sample, blood and tissue samples were taken. Group-2, rats underwent laminectomy. 0.9% saline with oral gavage (OG) done. On the 7th day, spinal cord sample, blood and tissue samples were taken. Group-3, rats underwent laminectomy. Spinal trauma was created with metal rods. With OG 0.9% saline was applied. On the 7th day, spinal cord sample, blood and tissue samples were taken. Group-4, rats underwent laminectomy. Spinal trauma was created with metal rods. with OG low dose myricetin (5mg/kg/day) was administered. 7th day spinal cord sample, blood and tissue samples were taken. Group-5 rats underwent laminectomy. Spinal trauma was created with metal rods. With OG high dose myricetin (25mg/kg/day) was administered. 7th day spinal cord sample, blood and tissue samples were taken. iv Basso Beattie and Bresnahan (BBB) for motor examinations of the rats on the 1st and 7th days of the experiment and Modified Tarlov tests were applied. Interleukin-1beta (IL1b) from damaged spinal cord tissue and blood obtained from rats,interleukin-10 (IL-10), total antioxidant status (TAS), total oxidative status (TOS), malonyldialdehyde (MDA) levels were determined by enzyme linked immunosorbent assay (ELISA) method our faculty Medical Biochemistry A.D. Worked in the Research Laboratory. Our Faculty Histology and Embryology A.D. spinal cord obtained from rats in the laboratorysections were examined in the histology laboratory using hematoxylin-eosin stain (H-E) examined under the light microscope. Apoptotic cells, ApopTag In Situ Apoptosis Detection Kit by TUNEL method (Millipore). TUNEL positive cells are 4 microns in gray matter. The percentages were taken by counting the sections with a 20 objective lens under the light microscope. Histopathology score and TUNEL positive cell count, one-way variance with TUKEY test statistical analysis was performed using ANOVA. Results: Myricetin caused an increase in IL-10 levels in serum and tissue independent of dose. While a decrease was observed in serum IL-1 beta levels independent of the drug dose, a significant decrease was observed in IL-1beta levels in the high-dose drug group in tissue samples. It was determined that when the application dose of myricetin increased, it created a more pronounced antioxidant response, causing a decrease in TOS level and an increase in TAS level. Again, when the application dose of Myricetin was increased, it was observed that MDA levels, which are strong markers of oxidative stress, caused a significant regression. In the histopathological examination, it was determined that TUNEL positive cell count and apoptotic index were significantly reduced by the effect of Myricetin. The lowest apoptotic index was determined in the high-dose drug group compared to the other experimental groups. Clinically, Basso-Beattie-Bresnahan (BBB) locomotor rating scale and Modified Tarloc Scale, post-traumatic Myricetin showed improvement in neurological examination. Conclusion: In the light of the biochemical, histopathological and neurological examination results we obtained, myricetin has shows anti-inflammatory, antioxidant, antiapoptotic and neuroprotective activity on secondary spinal cord injury.
Objective : In this project, we aimed to identify a class of flavonoid polyphenolic compounds with antioxidant properties. By applying Myricetin, which is a member of the family, to rats with traumatic spinal cord injury. Myricetin has a secondary effect on free radicals in the spinalcord after trauma. We evaluated its neuroprotective effects on injury. Method: 45 Wistar albino rats were randomized into 5 groups, 9 in each group was done. Traumatic spinal cord injury, weight reduction model with titanium rodscarried out. Group-1 (sham), rats underwent laminectomy. 7th day spinal cord sample, blood and tissue samples were taken. Group-2, rats underwent laminectomy. 0.9% saline with oral gavage (OG) done. On the 7th day, spinal cord sample, blood and tissue samples were taken. Group-3, rats underwent laminectomy. Spinal trauma was created with metal rods. With OG 0.9% saline was applied. On the 7th day, spinal cord sample, blood and tissue samples were taken. Group-4, rats underwent laminectomy. Spinal trauma was created with metal rods. with OG low dose myricetin (5mg/kg/day) was administered. 7th day spinal cord sample, blood and tissue samples were taken. Group-5 rats underwent laminectomy. Spinal trauma was created with metal rods. With OG high dose myricetin (25mg/kg/day) was administered. 7th day spinal cord sample, blood and tissue samples were taken. iv Basso Beattie and Bresnahan (BBB) for motor examinations of the rats on the 1st and 7th days of the experiment and Modified Tarlov tests were applied. Interleukin-1beta (IL1b) from damaged spinal cord tissue and blood obtained from rats,interleukin-10 (IL-10), total antioxidant status (TAS), total oxidative status (TOS), malonyldialdehyde (MDA) levels were determined by enzyme linked immunosorbent assay (ELISA) method our faculty Medical Biochemistry A.D. Worked in the Research Laboratory. Our Faculty Histology and Embryology A.D. spinal cord obtained from rats in the laboratorysections were examined in the histology laboratory using hematoxylin-eosin stain (H-E) examined under the light microscope. Apoptotic cells, ApopTag In Situ Apoptosis Detection Kit by TUNEL method (Millipore). TUNEL positive cells are 4 microns in gray matter. The percentages were taken by counting the sections with a 20 objective lens under the light microscope. Histopathology score and TUNEL positive cell count, one-way variance with TUKEY test statistical analysis was performed using ANOVA. Results: Myricetin caused an increase in IL-10 levels in serum and tissue independent of dose. While a decrease was observed in serum IL-1 beta levels independent of the drug dose, a significant decrease was observed in IL-1beta levels in the high-dose drug group in tissue samples. It was determined that when the application dose of myricetin increased, it created a more pronounced antioxidant response, causing a decrease in TOS level and an increase in TAS level. Again, when the application dose of Myricetin was increased, it was observed that MDA levels, which are strong markers of oxidative stress, caused a significant regression. In the histopathological examination, it was determined that TUNEL positive cell count and apoptotic index were significantly reduced by the effect of Myricetin. The lowest apoptotic index was determined in the high-dose drug group compared to the other experimental groups. Clinically, Basso-Beattie-Bresnahan (BBB) locomotor rating scale and Modified Tarloc Scale, post-traumatic Myricetin showed improvement in neurological examination. Conclusion: In the light of the biochemical, histopathological and neurological examination results we obtained, myricetin has shows anti-inflammatory, antioxidant, antiapoptotic and neuroprotective activity on secondary spinal cord injury.
Açıklama
Anahtar Kelimeler
Deneysel, Myricetin, Rat, Spinal kord, Travma, Experimental, Myricetin, Rat, Secondary injury, Spinal cord, Trauma
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Kaplan, S. (2023). Deneysel akut spinal kord hasarında myrıcetın'in nöroprotektif etkilerinin değerlendirilmesi. (Yayınlanmamış tıpta uzmanlık tezi) Necmettin Erbakan Üniversitesi, Meram Tıp Fakültesi Cerrahi Tıp Bilimleri Bölümü Beyin ve Sinir Cerrahisi Anabilim Dalı, Konya.
