Geçici fokal serebral iskemi uygulanan sıçanlarda ardkoşullama uygulamasının lipid peroksidasyonu ve apopitozis üzerine etkisi
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Date
2007
Authors
Abaş, Faruk
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Journal ISSN
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Publisher
Uludağ Üniversitesi
Abstract
İskemiyi takiben reperfüzyonun erken döneminde kan akımının hızlı ve aralıklı olarak kesintiye uğramasına ardkoşullama denir. Çalışmamızın amacı, geçici fokal serebral iskemi modeli uygulanan sıçanlarda ard koşullama uygulamasının lipid peroksidasyonu ve apoptozis üzerine etkisinin araştırılmasıdır. Geçici fokal serebral iskemi, 32 adet (275±25g) Spraque-Dawley erkek sıçanlarda intraluminal filament yöntemi ile orta serebral arterde geçici oklüzyon (OSAo) oluşturuldu. OSAo doğrulanması ipsilateral parietal kortekste Laser Doppler Flowmetre monitörizasyonu ve takiben nörolojik muayene ile yapıldı. Deney grupları; iskemi ve ardkoşullama grubu olarak planlandı. 120 dakikalık oklüzyonu takiben 24 saat reperfüze edilerek dekapite edildi. İmmunohistokimyasal analiz için antiapoptotik protein olan bcl-2, proapoptotik olan kaspaz 3, kaspaz 9 ve bax immun boyamaları yapıldı. Lipid peroksidasyonu saptamak içinde her iki hemisferde dokuda malonildialdehit (MDA) seviyeleri saptandı. Tüm sıçanlarda serebral kan akımı değerlerinde anlamlı düşüş saptandı (p<0.05). İskemik hemisferle normal hemisferler arasında her iki grupta istatiksel olarak anlamlı düzeyde MDA seviyelerinde artış saptandı (p<0.05). Grup içi iskemik hemisfer değerleri normal hemisfer karşılaştırıldığında ardkoşullama yapılan grupta MDA seviyelerinde yüzde değişim açısından %10.81’lik artış saptanırken, iskemi oluşturulan grupta %27.81’lik artış saptandı ve bu değişim istatistiksel olarak anlamlı bulundu (p<0.05). Ardkoşullama grubunda bcl-2 immunohistokimyasal skorlarında iskemi grubu ile karşılaştırıldığında artış saptandı (p<0.05). Kaspaz 3, kaspaz 9 ve bax immun boyamaları açısından gruplar arası istatiksel farklılık saptanmadı. Sonuç olarak ardkoşullamanın, lipid peroksidasyonu ve apoptozis üzerinden etki göstererek serebral iskemiden koruduğu düşünülmektedir.
Postconditioning is defined as rapid and transient blood flow interruption in the early phase of reperfusion. In our study, we aimed to investigate the effects of postconditioning on the lipid peroxidation and apoptosis in the rat models of transient focal cerebral ischemia. Transient focal cerebral ischemia was produced by intraluminal filament method in 32 Spraque-Dawley male rats (275±25g). Adequacy of MCA occlusion was monitored with Laser Doppler flowmetry over the ipsilateral cortex and tested with post procedural neurological examination. The groups were planned as ischemia and postconditioning. Rats were decapitated after 24 hours reperfusion following 120 minutes occlusion. Immunohistochemical stainings of bcl-2 as antiapoptotic protein, caspase 3, caspase 9 and bax as proapoptotic protein were performed. Determination of the lipid peroxidation of malonyldi-aldehyde (MDA) was measured in both of the cerebral hemispheres. After MCA occlusion, there was a significant drop in the CBF in all rats (p<0.05). In groups, there was significant increase in the MDA levels when normal and ischemic hemispheres were compared (p<0.05). When MDA ingroup values of ischemic hemisphere were compared with normal hemisphere (100%); 27.81% and 10.81% increases were found in the ischemia group and the postconditioning group, respectively and this was statistically significant (p<0.05). When postconditioning group was compared with ischemia group, there was a statistically significant increase in bcl-2 immunohistochemical scores (p<0.05). There was no statistical significant change between groups in terms of caspase 3, caspase 9 and bax immune staining. As a result, we suggested that postconditioning was effective against cerebral ischemia through lipid peroxidation and apoptosis.
Postconditioning is defined as rapid and transient blood flow interruption in the early phase of reperfusion. In our study, we aimed to investigate the effects of postconditioning on the lipid peroxidation and apoptosis in the rat models of transient focal cerebral ischemia. Transient focal cerebral ischemia was produced by intraluminal filament method in 32 Spraque-Dawley male rats (275±25g). Adequacy of MCA occlusion was monitored with Laser Doppler flowmetry over the ipsilateral cortex and tested with post procedural neurological examination. The groups were planned as ischemia and postconditioning. Rats were decapitated after 24 hours reperfusion following 120 minutes occlusion. Immunohistochemical stainings of bcl-2 as antiapoptotic protein, caspase 3, caspase 9 and bax as proapoptotic protein were performed. Determination of the lipid peroxidation of malonyldi-aldehyde (MDA) was measured in both of the cerebral hemispheres. After MCA occlusion, there was a significant drop in the CBF in all rats (p<0.05). In groups, there was significant increase in the MDA levels when normal and ischemic hemispheres were compared (p<0.05). When MDA ingroup values of ischemic hemisphere were compared with normal hemisphere (100%); 27.81% and 10.81% increases were found in the ischemia group and the postconditioning group, respectively and this was statistically significant (p<0.05). When postconditioning group was compared with ischemia group, there was a statistically significant increase in bcl-2 immunohistochemical scores (p<0.05). There was no statistical significant change between groups in terms of caspase 3, caspase 9 and bax immune staining. As a result, we suggested that postconditioning was effective against cerebral ischemia through lipid peroxidation and apoptosis.
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Keywords
Geçici fokal serebral iskemi, Ardkoşullama, Apopitozis, Lipid peroksidasyonu, Transient focal cerebral ischemia, Postconditioning, Lipid peroxidation, Apoptosis
Citation
Abaş, F. (2007). Geçici fokal serebral iskemi uygulanan sıçanlarda ardkoşullama uygulamasının lipid peroksidasyonu ve apopitozis üzerine etkisi. Yayınlanmamış uzmanlık tezi. Uludağ Üniversitesi Tıp Fakültesi.