Ahmet ALTUN, Mesut PARLAK, Tijen TEMİZ KAYA, Zeliha ÖDEMİŞ BEŞİKÇİ, Ezgi BAlCI

Simvastatin ve mevastatin'in anjiogenez inhibisyonu üzerine etkilerinin koryoallantoik membran modelinde araştırılması

Amaç. Bu çalışmanın amacı klinik tedavide sık olarak kullanılan statinlerdenmevastatinin anjiogenez inhibisyonu üzerine etkilerini döllenmiş tavuk yumurtaları üzerinde,koryoallantoik membran modelini (CAM modeli) kullanarak kıyaslamaktır. Yöntem. ÇalışmadaRoss 308 cinsi döllenmiş tavuk yumurtaları kullanıldı. Döllenmiş tavuk yumurtaları 37,5°C‟de%80 rölatif nemli ortamda horizontal pozisyonda inkübe edildi. Kuluçkanın beşinci günündeyumurtanın künt tarafından enjektör yardımıyla 5 mL albumin alındı ve yumurtanın diğer ucundan2-3 cm çapında kabuk kesilerek çıkarıldı. Kabuktaki bu açıklık laboratuvar filmi ile kapatıldı vekoryoallantoik membran modeli yaklaşık 2 cm çapa ulaşana kadar 72 saat daha inkübe edildi. Herbir yumurtaya koryoallantoik membran üzerine etken madde içeren bir pellet yerleştirildi. İlaçuygulamasından (bevacizumab, simvastatin ve mevastatin; 10-4M, 10-5M ve 10-6Mkonsantrasyon) sonra yumurtalar 24 saat daha inkübe edildi. Stereoskopik mikroskop altındaBürgermeister ve arkadaşlarının skorlama sistemi kullanılarak pellet uygulama bölgesindeki damaryapısı değerlendirildi. Bulgular. Çalışmamızda negatif kontrol olarak ilaç içermeyen agar disklerkullanıldı. Negatif kontrol yumurtalarının hiç birinde anjiogenez engellenmedi ve anti-anjiogeniketki puanı 0 olarak bulundu. Pozitif kontrol olarak antianjiogenik etkinliği kanıtlanmışbevacizumab kullanıldı. Bevacizumab‟ın 10-4M,10-5M ve 10-6M konsantrasyonlarındaantianjiogenetik skor değerleri sırasıyla 1,58 , 1,55 ve 1,00 olarak bulundu. Simvastatinin vemevastatinin 10-4M, 10-5M ve 10-6M konsantrasyonları için ortalama antianjiyogenik skordeğerleri sırası ile 0,93 , 0,66 , 0,53 ve 1,0, 0,80 ve 0,66 olarak bulundu. Hem simvastatinin hemde mevastatinin her üç konsantrasyonunun da anti anjiogenik etkiye sahip olduğu saptandı. Bu ikiilacın 10-4M konsantrasyonlarının meydana getirmiş olduğu anti anjiogenik etki gücü arasındaanlamlı bir fark bulunamadı. Ancak azalan konsantrasyonlar birbirleri ile karşılaştırıldığındamevastatinin 10-5M ve 10-6 M konsantrasyonlarda simvastatine göre daha anlamlı bir antianjiogenik etki meydana getirdiği saptandı. Sonuç. Sonuç olarak simvastatin ve mevastatin CAMmodelinde antiantiogenik etkinlik gösteren statinlerdir. Bu iki statin lipid düşürücü etkileriyanında, anjiogenez inhibisyonu yaparak; plak içi antianjiogenik etkiyle, plak rüptürü ve bunabağlı ciddi komplikasyonlardan da korunma sağlayabilir.

Investigating the effects of simvastatin and mevastatin on angiogenesis in chorioallantoic membrane model

Aim. The purpose of this study is to compare the effects of Simvastatin and Mevastatin,commonly used statins in clinic practice, on angiogenesis in chorioallontioc membrane models byusing fertilized chicken eggs. Methods. In this study, Ross 308, fertilized chicken eggs were used.The fertilized chicken eggs were incubated in horizontal position with environmental conditions of37.5 °C temperature and 80% relative humidity. On the fifth day of the incubation period, 5 ml ofalbumen was taken through the eggshell with a syringe and a shell piece of 2-3 cm in diameter wasremoved from the contrary side of the eggs. The windows on the egg shells were sealed withgelatin and thereafter, the eggs were incubated for 72 more hours to have chorioallontiocmembrane models reaching 2 cm in diameter. The pellets containing active substrate were placedon the chorioallantoic membrane of each egg. After drug administration (bevacizumab,Simvastatin and Mevastatin; 10-4M, 10-5M ve 10-6M concentration), the eggs were incubated for24 hours again. Under a stereoscopic microscope, vascular structure around the pellets wasevaluated using a scoring system of Bürgermeister et al. Results. Drug-free agar discs were usedas negative control in our study. Angiogenesis was not inhibited in any negative control eggs andanti-angiogenic effect score was found as 0 .Bevacizumab was used as positive control. In 10-4 M, 10-5M and 10-6M concentrations, antiangiogenic score of Bevacizumab were found 1,58, 1,55and 1,00, respectively. For 10-4M, 10-5M and 10-6M concentrations of Simvastatin andMevastatin, antiangiogenic scores were found as 0.93, 0.66, 0.53 and 1.0, 0.80, 0.66, respectively.Both Mevastatin and Simvastatin were found to have antiangiogenic effects in each threeconcentrations. There wasn‟t a significant difference between antiangiogenic effects of these twodrugs in 10-4M. But when lower concentrations were compared it is found that Mevastatin hadsignificantly higher antiangiogenic effect than Simvastatin at 10-5M and 10-6M concentrations.Conclusion. In conclusion, Simvastatin and Mevastatin are statins which shows antiangiogeniceffect in chorioallontioc membrane models. Besides they can the lipid-lowing effects of statinsthey can also provide. Protection against plaque rupture related serious complication by theintraplaque antiangiogenesis effect

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