The Effect of Ca-074 (Cathepsın B Inhibitor) on Necrotic and Apoptotic Neuronal Cell Death in Model of Cerebral Ischemia

Yıl 2023, Cilt: 45 Sayı: 5, 781 - 790, 27.09.2023

Emre Özkara , Ramazan Durmaz , Zühtü Özbek , Hilmi Özden , Güngör Kanbak , Kubilay Uzuner

https://doi.org/10.20515/otd.1343599

Öz

Lysosomes and cathepsins, the most common hydrolytic enzymes in lysosomes, are available in the different models of cell death as necrosis and apoptosis. This study investigated the effect of cathepsin B-selective inhibitor CA-074 on apoptotic and necrotic neuronal cell death. Focal cerebral ischemia which has been formed by occlusion of the three-vessel consisting permanent middle cerebral artery occlusion and temporary bilateral common carotid artery occlusion for 60 minutes was selected as ischemia model. Two sets of rats were used in this study. The rats in the first set were used formeasurement of sulfhydryl groups in the lysosomal membrane, lysosomal integrity, cathepsins B and L activities and caspase-3 activity. The rats in the second set were used as histological study including "hematoxylin and eosin" for the detection of necrotic neuronal deathand "TUNEL" staining for the detection of apoptotic neuronal death. 4 mg/kg CA-074 was administered intravenouslyin the treatment group. CA-074 has substantially reduced levels of cathepsins B and L compared to ischemia and solvent groups (respectively, p<0.05 and p<0.01). Similarly, CA-074 has reduced increase in caspase-3 activity compared to ischemia and solvent groups (p<0.05). While the number of eosinophilic (necrotic) and apoptotic neurons has highly increased in post-ischemic cerebral tissue in middle cerebral artery feeding area (p<0.001), CA-074 could only reduce significantly the number of apoptotic neurons (p<0.05). CA-074 has reduced apoptotic neuronal death by inhibiting caspase and cathepsin activity. It may be useful that CA074 is used with other therapeutic drugs in stroke patients.

Anahtar Kelimeler

ischemia, cerebral, rat model, CA074

Katepsin B İnhibitörü olan CA074'ün Serebral İskemi Modelinde Apoptoz ve Hücre Ölümü Üzerine Etkisi

Öz

Lizozomlarda en yaygın hidrolitik enzimlerden olan lizozomlar ve katepsinler, nekroz ve apoptoz olarak farklı hücre ölümü modellerinde görev almaktadır. Bu çalışma, katepsin B-seçici inhibitörü olan CA-074'ün apoptotik ve nekrotik nöronal hücre ölümü üzerindeki etkisini araştırdı. Bu çalışmda İskemi modeli olarak kalıcı orta serebral arterin tıkanıklığı ve geçici bilateral ana karotid arter tıkanıklığından oluşan üç damarın 60 dakika süreyle oklüzyonu ile oluşturulan fokal serebral iskemi modeli seçilmiştir. Çalışmada iki set sıçan kullanıldı. Birinci setteki ratlar lizozomal membrandaki sülfhidril gruplarının, lizozomal bütünlüğün, katepsin B ve L aktivitelerinin ve kaspaz-3 aktivitesinin ölçülmesinde kullanıldı. İkinci setteki sıçanlar, nekrotik nöronal ölümün saptanması için "hematoksilen ve eozin" ve apoptotik nöronal ölümün saptanması için "TUNEL" boyamasını içeren histolojik çalışma olarak kullanıldı. Tedavi grubuna 4 mg/kg CA-074 intravenöz olarak uygulandı. CA-074, iskemi ve solvent gruplarına kıyasla önemli ölçüde azalmış katepsin B ve L seviyelerine sahiptir (sırasıyla, p<0.05 ve p<0.01). Benzer şekilde CA-074 kaspaz-3 aktivitesindeki artışı iskemi ve solvent gruplarına göre azaltmıştır (p<0.05). Orta serebral arter beslenme alanındaki iskemik sonrası serebral dokuda eozinofilik (nekrotik) ve apoptotik nöronların sayısı oldukça artarken (p<0.001), CA-074 sadece apoptotik nöronların sayısını önemli ölçüde azaltabildi (p<0.05). CA-074, kaspaz ve katepsin aktivitesini inhibe ederek apoptotik nöronal ölümü azaltmıştır. CA074'ün inme hastalarında diğer terapötik ilaçlarla birlikte kullanılması faydalı olabilir.

Anahtar Kelimeler

iskemi, beyin, rat modeli, CA074

Destekleyen Kurum

Eskişehir Osmangazi Üniversitesi

Kaynakça

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