Ratlarda deneysel subaraknoid hemorajide wogonin'in serebral vazospazma yönelik iyileştirici özellikleri
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Tarih
2024
Yazarlar
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Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Bu çalışmada Wogonin’in subaraknoid kanama (SAK) sonrası görülen serebral vazospazm üzerine iyileştirici etkilerini incelemek amaçlanmaktadır.
Gereç ve Yöntem: Wistar Albino cinsi 250-350 gram ağırlığında toplam 60 adet rat çalışmaya dahil edildi. Deney grupları, Grup 1 (kontrol, n=12), Grup 2 (Sham grubu, n=12), Grup 3 (SAK Grubu, n=12), Grup 4 (SAK+DMSO grubu, n=12), Grup 5 (SAK+Wogonin grubu, n=12) olacak şekilde randomize edildi. Subaraknoid kanama, sisterna magnaya çift enjeksiyon modeliyle gerçekleştirildi. Deneyin başlangıcından ratların sakrifikasyonuna kadar ilk gün 0, son gün 7 olacak şekilde günler numaralandırıldı. SAK indüksiyonu çift enjeksiyon modeli ile deneyin 0. ve 2. günlerinde uygulandı. Wogonin, çift enjeksiyon modeli ile SAK indüksiyonu tamamlandıktan hemen sonra intraperitoneal yolla 40 mg/kg dozunda uygulandı. Anestezi, cerrahi işlemlerin öncesinde 10 mg/kg xylazine ve 60 mg/kg ketamin ile sağlandı. Deneyin 7. günü intrakardiyak kan ve serebral doku örnekleri alınarak hayvanlar sakrifiye edildi. Elde edilen santral sinir sistemi dokusunda ve kanda interlökin 1 Beta (IL-1β), interlökin 10 (IL-10), sinir büyüme faktörü (NGF), glial derived büyüme faktörü (GDNF) seviyeleri enzyme linked immunosorbent assay (ELİSA) yöntemiyle ve total antioksidan stres (TAS), total oksidan stres (TOS) seviyeleri spektrofotometri yöntemiyle çalışıldı. Baziller arter görülecek şekilde transvers kesilerek bloklanan doku örnekleri, Hematoksilen Eozin ile boyandı. Preparatların morfometrik olarak, görüntülü analiz programı eşliğinde ve ışık mikroskobu altında damar çapları ölçüldü. İmmünhistokimyasal Apoptoz boyama (TUNNEL Metodu) ile vasküler endoteldeki ekpresyon yüzdeleri tespit edildi. Sayısal değişkenler için ortalama ve standart sapma istatistikleri verildi. Sayısal değişkenlerin analizinde ANOVA ve karma etki modellerinden (mixed effects models) faydalanıldı. Posthoc ikili karşılaştırmalarda Tukey Testi ya da Tukey düzeltmeli en küçük kare ortalama karşılaştırmaları yapıldı. Verilerin analizi R 4.4.1 (R Core Team, 2024) programi ile yapıldı. p<0,05 istatistiksel olarak anlamlı kabul edildi.
Bulgular: Bu çalışmada yapılan histopatolojik incelemede apoptozisi gösteren ortalama TUNEL pozitif endotel hücre yüzdesinin SAK grubunda, Kontrol grubuna göre istatistiksel açıdan anlamlı miktarda yüksek olduğu görüldü. Wogonin tedavisi ile ortalama TUNEL pozitif hücre yüzdesinin istatistiksel olarak anlamlı düzeyde azaldığı gözlendi. SAK grubu ortalama baziller arter çapı değerlerinin kontrol grubuna göre anlamlı düzeyde azaldığı, Wogonin uygulanan grupta ise bu azalmanın olmadığı görüldü. Wogonin grubunda, inflamatuar sitokin olan IL-1β düzeyinde SAK grubuna göre anlamlı azalma gözlendi. Wogonin tedavisi nöroprotektif etkileri gösteren NGF ve GDNF düzeylerinde SAK grubuna göre anlamlı artış sağlamıştır. Ayrıca SAK nedeniyle ortalama TOS düzeylerinde görülen artışın ve ortalama TAS düzeylerinde görülen azalmanın, Wogonin uygulanan grupta anlamlı şekilde düzeldiği görüldü.
Sonuç: Wogonin tedavisinin etkilerini araştıran bu deneysel çalışmada, Wogonin’in SAK sonrası artan endotelyal apoptozisi azalttığı ve azalan baziller arter çapını artırdığı gözlendi. Ayrıca yapılan biyokimyasal incelemelerde Wogonin’in SAK sonrası gelişen serebral vazospazmda antiinflamatuar, nöroprotektif ve antioksidan etkileri olduğu gösterildi.
Aim: The aim of this study is to investigate the ameliorative effects of Wogonin on cerebral vasospasm after subarachnoid hemorrhage (SAH). Material and Method: A total of 60 Wistar Albino rats weighing 250-350 grams were included in the study. The experimental groups were randomized as Group 1 (control, n=12), Group 2 (Sham group, n=12), Group 3 (SAH group, n=12), Group 4 (SAH+DMSO group, n=12), Group 5 (SAH+Wogonin group, n=12). Subarachnoid hemorrhage was performed with a double injection model into the cisterna magna. From the beginning of the experiment until the sacrification of the rats, the days were numbered as 0 on the first day and 7 on the last day. SAH induction was performed on days 0 and 2 of the experiment with the double injection model. Wogonin was administered intraperitoneally at a dose of 40 mg/kg immediately after SAH induction was completed with the double injection model. Anesthesia was induced with 10 mg/kg xylazine and 60 mg/kg ketamine before surgical procedures. On the 7th day of the experiment, intracardiac blood and cerebral tissue samples were obtained and the animals were sacrificed. Interleukin 1 Beta (IL-1β), interleukin 10 (IL-10), nerve growth factor (NGF), glial derived growth factor (GDNF) levels were determined by enzyme linked immunosorbent assay (ELISA) method and total antioxidant stress (TAS), total oxidant stress (TOS) levels were determined by spectrophotometry method in the obtained central nervous system tissue and blood. The tissue samples were transversely cut and blocked to visualize the basilar artery and stained with Hematoxylin Eosin. The tissue specimens were transversely cut and blocked to visualize the basilar artery and stained with Hematoxylin Eosin. Vascular diameters of the preparations were measured morphometrically under a light microscope and with the help of an image analysis pram. Immunohistochemical Apoptosis staining (TUNNEL Method) was used to determine the expression percentages in the vascular endothelium. Mean and standard deviation statistics were given for numerical variables. ANOVA and mixed effects models were used in the analysis of numerical variables. Posthoc pairwise comparisons were performed using Tukey's test or least square mean comparisons with Tukey's correction. Data analysis was performed with R 4.4.1 (R Core Team, 2024). p<0.05 was considered statistically significant. Results: In the histopathological examination performed in this study, the mean percentage of TUNEL positive endothelial cells indicating apoptosis was statistically significantly higher in the SAH group than in the control group. A statistically significant decrease in the mean TUNEL positive cell percentage was observed with wogonin treatment. Mean basilar artery diameter values in the SAH group were significantly decreased compared to the control group, whereas this decrease was not observed in the Wogonin-treated group. A significant decrease was observed in the level of inflammatory cytokine IL-1β in the Wogonin group compared to the SAH group. Wogonin treatment provided a significant increase in NGF and GDNF levels showing neuroprotective effects compared to the SAH group. In addition, the increase in mean TOS levels and decrease in mean TAS levels due to SAH were significantly improved in the Wogonin-treated group. Conclusion: In this experimental study investigating the effects of Wogonin treatment, it was observed that Wogonin decreased endothelial apoptosis and increased basilar artery diameter after SAH. In addition, biochemical analyses showed that Wogonin had anti-inflammatory, neuroprotective and antioxidant effects on cerebral vasospasm after SAH.
Aim: The aim of this study is to investigate the ameliorative effects of Wogonin on cerebral vasospasm after subarachnoid hemorrhage (SAH). Material and Method: A total of 60 Wistar Albino rats weighing 250-350 grams were included in the study. The experimental groups were randomized as Group 1 (control, n=12), Group 2 (Sham group, n=12), Group 3 (SAH group, n=12), Group 4 (SAH+DMSO group, n=12), Group 5 (SAH+Wogonin group, n=12). Subarachnoid hemorrhage was performed with a double injection model into the cisterna magna. From the beginning of the experiment until the sacrification of the rats, the days were numbered as 0 on the first day and 7 on the last day. SAH induction was performed on days 0 and 2 of the experiment with the double injection model. Wogonin was administered intraperitoneally at a dose of 40 mg/kg immediately after SAH induction was completed with the double injection model. Anesthesia was induced with 10 mg/kg xylazine and 60 mg/kg ketamine before surgical procedures. On the 7th day of the experiment, intracardiac blood and cerebral tissue samples were obtained and the animals were sacrificed. Interleukin 1 Beta (IL-1β), interleukin 10 (IL-10), nerve growth factor (NGF), glial derived growth factor (GDNF) levels were determined by enzyme linked immunosorbent assay (ELISA) method and total antioxidant stress (TAS), total oxidant stress (TOS) levels were determined by spectrophotometry method in the obtained central nervous system tissue and blood. The tissue samples were transversely cut and blocked to visualize the basilar artery and stained with Hematoxylin Eosin. The tissue specimens were transversely cut and blocked to visualize the basilar artery and stained with Hematoxylin Eosin. Vascular diameters of the preparations were measured morphometrically under a light microscope and with the help of an image analysis pram. Immunohistochemical Apoptosis staining (TUNNEL Method) was used to determine the expression percentages in the vascular endothelium. Mean and standard deviation statistics were given for numerical variables. ANOVA and mixed effects models were used in the analysis of numerical variables. Posthoc pairwise comparisons were performed using Tukey's test or least square mean comparisons with Tukey's correction. Data analysis was performed with R 4.4.1 (R Core Team, 2024). p<0.05 was considered statistically significant. Results: In the histopathological examination performed in this study, the mean percentage of TUNEL positive endothelial cells indicating apoptosis was statistically significantly higher in the SAH group than in the control group. A statistically significant decrease in the mean TUNEL positive cell percentage was observed with wogonin treatment. Mean basilar artery diameter values in the SAH group were significantly decreased compared to the control group, whereas this decrease was not observed in the Wogonin-treated group. A significant decrease was observed in the level of inflammatory cytokine IL-1β in the Wogonin group compared to the SAH group. Wogonin treatment provided a significant increase in NGF and GDNF levels showing neuroprotective effects compared to the SAH group. In addition, the increase in mean TOS levels and decrease in mean TAS levels due to SAH were significantly improved in the Wogonin-treated group. Conclusion: In this experimental study investigating the effects of Wogonin treatment, it was observed that Wogonin decreased endothelial apoptosis and increased basilar artery diameter after SAH. In addition, biochemical analyses showed that Wogonin had anti-inflammatory, neuroprotective and antioxidant effects on cerebral vasospasm after SAH.
Açıklama
Anahtar Kelimeler
Subaraknoid Kanama, Subarachnoid Haemorrhage, Vazospazm, Vasospasm, Wogonin, Rat
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Yüksek, M. E. (2024). Ratlarda deneysel subaraknoid hemorajide wogonin'in serebral vazospazma yönelik iyileştirici özellikleri. (Yayınlanmamış tıpta uzmanlık tezi) Necmettin Erbakan Üniversitesi, Tıp Fakültesi Cerrahi Tıp Bilimleri Bölümü Beyin ve Sinir Cerrahisi Anabilim Anabilim, Konya.
