miR-17/20’nin Sıçanların Subaraknoid Kanama Modelinde Gelişen Vazospazm Üzerine Etkisi

Year 2022, , 256 - 262, 31.12.2022

Başak Büyük , Ümit Ali Malçok

https://doi.org/10.30565/medalanya.1152279

Abstract

Amaç: Mevcut çalışmamızın amacı, ratlarda oluşturulan subaraknoid kanama (SAH) modelinde melatonin ve miRNA-17/20 uygulamasının basiller arterde gelişen vazospazm ve damar hasarı üzerine etkisinin araştırılmasıdır.

Yöntemler: Çalışmada kullanılan 36 adet sıçan 6 gruba ayrılmıştır; Sham, SAH, SAH+NegmiRNA, SAH+MEL, SAH-miRs-17/20 group, SAH+MEL+miRs-17/20. Sıçanlara anestezi altında SAH modeli uygulandı. Kafanın ön bölgesinden dikey bir kesi ile cilt açılarak kemiğe ulaşıldı. İnsizyondan sonra, kuyruk arterinden toplanan 120 µL heparinize olmayan taze otolog arteriyel kan aseptik koşullar altında 10 saniyede yavaşça prekiazmatik sisternaya enjekte edildi. Sham grubunda kan enjeksiyonu dışında tüm basamaklar SAH grubuyla aynıydı. SAH+NegmiRs-17/20 grubunda, miRs-17/20 miRNA Mimic Negative Control#1 SAH operasyonundan 1 saat sonra uygulandı. SAH+MEL grubunda, SAH operasyonundan 1 saat sonra 10 mg/kg melatonin intraperitoneal olarak verildi. SAH-miRs-17/20 grubundaysa SAH operasyonundan 1 saat sonra intranazal olarak mimic-miR-17 ve mimic-miR-20 verildi. SAH+MEL+miRs-17/20 grubunda, SAH operasyonundan 1 saat sonra intranazal mimic-miR-17 ile intraperitoneal melatonin (10 mg/kg) uygulandı. Deney sonunda sıçanlardan basiller arteri de içeren beyin örnekleri alınıp rutin doku takip işlemlerine tabi tutuldu. Sonrasında H-E yöntemi ile boyanan histolojik kesitlerde damar örnekleri vazospazm, tunica mediada ödem ve lamina elastica internada kıvrımlanma kriterleri açısından değerlendirilip derecelendirildi.

Bulgular: SAH modeli oluşturulan sıçanlarda, SAH modeli sonrası melatonin ve miRs-17/20’nin birlikte uygulanmasının damar duvarında oluşan vazospazm ve ödem oluşmasını anlamlı şekilde azalttığı gösterilmiştir. SAH sonrası miRs-17/20 uygulamasının da tek başına damar duvarında ödem gelişmesini ve vazospazma bağlı lamina elastica internada kıvrımlanmayı azalttığı ortaya konmuştur.

Sonuç: Çalışmamızda, miRs-17/20’nin damar duvarında vazospazmı azaltabileceği, ödemi de azaltarak damar hasarını önleyebileceği gösterilmiştir.

Keywords

Subaraknoid kanama, microRNA, melatonin, sıçan

Effect of miRs-17/20 on vasospasm in subarachnoid hemorrhage model of rats

Abstract

Aim: To investigate the effects of melatonin and miRNA-17/20 administration on vasospasm and vascular damage on the bacillary artery in the Subarachnoid hemorrhage (SAH) model of rats.

Methods: Rats were divided into 6 groups: Sham, SAH, SAH+NegmiRNA, SAH+MEL, SAH-miRs-17/20 group, SAH+MEL+miRs-17/20. For creating the SAH model the skin was cut with a vertical incision in the anterior region of the head.120 µL of fresh non-heparinized autologous arterial blood collected from the tail artery was injected into the prechiasmatic cistern under aseptic conditions. All steps in the Sham were the same as in the SAH group, except for blood injection. In the SAH+NegmiRs-17/20, miRs-17/20 miRNA Mimic-Negative Control#1 was administered 1 hour after SAH operation. In the SAH+MEL,10 mg/kg melatonin was administered intraperitoneally 1 hour after the SAH operation. In the SAH-miRs-17/20, mimic-miR-17 and mimic-miR-20 were given intranasally 1 hour after the SAH operation. In the SAH+MEL+miRs-17/20,intranasal mimic-miR-17 and intraperitoneal melatonin were administered 1 hour after the SAH operation. Brain samples, including the bacillary artery, were taken and subjected to routine tissue processing procedures. Vessel samples were evaluated and graded in histological sections stained with the H-E method in terms of vasospasm, edema in the tunica media, and folding of the lamina elastica interna.

Results: The co-administration of melatonin and miRs-17/20 reduced the vasospasm and edema formation in the vessel wall. It has also been demonstrated that the application of miRs-17/20 after SAH alone reduces the development of edema in the vessel wall and folding of the internasal lamina elastica due to vasospasm.

Conclusion: It has been shown that miRs-17/20 can reduce vasospasm in the vessel wall and prevent vessel damage by reducing edema.

Keywords

Subarachnoid Hemorrhage, microRNAs, rats, melatonin,

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