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

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

Makale detayları

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

Yazarlar:

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

Yayıncı:

Eskişehir Osmangazi University

Ana Başlıklar:

Beyin ve Sinir Cerrahisi (Nöroşirurji)

Dergi:

Osmangazi Tıp Dergisi Vol.45 Issue 5, pp.781-790

Belge Tipi:

Makale

Yayın Tarihi:

9/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001293346

pISSN:

1305-4953

Anahtar kelimeler:

Özet:

 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 mgkg 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, p0.05 and p0.01. Similarly, CA-074 has reduced increase in caspase-3 activity compared to ischemia and solvent groups p0.05. While the number of eosinophilic necrotic and apoptotic neurons has highly increased in post-ischemic cerebral tissue in middle cerebral artery feeding area p0.001, CA-074 could only reduce significantly the number of apoptotic neurons p0.05. 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.

 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 mgkg 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, p0.05 and p0.01. Similarly, CA-074 has reduced increase in caspase-3 activity compared to ischemia and solvent groups p0.05. While the number of eosinophilic necrotic and apoptotic neurons has highly increased in post-ischemic cerebral tissue in middle cerebral artery feeding area p0.001, CA-074 could only reduce significantly the number of apoptotic neurons p0.05. 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.

 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 mgkg 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, p0.05 and p0.01. Similarly, CA-074 has reduced increase in caspase-3 activity compared to ischemia and solvent groups p0.05. While the number of eosinophilic necrotic and apoptotic neurons has highly increased in post-ischemic cerebral tissue in middle cerebral artery feeding area p0.001, CA-074 could only reduce significantly the number of apoptotic neurons p0.05. 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.

 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-074ün 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 mgkg CA-074 intravenöz olarak uygulandı. CA-074, iskemi ve solvent gruplarına kıyasla önemli ölçüde azalmış katepsin B ve L seviyelerine sahiptir sırasıyla, p0.05 ve p0.01. Benzer şekilde CA-074 kaspaz-3 aktivitesindeki artışı iskemi ve solvent gruplarına göre azaltmıştır p0.05. Orta serebral arter beslenme alanındaki iskemik sonrası serebral dokuda eozinofilik nekrotik ve apoptotik nöronların sayısı oldukça artarken p0.001, CA-074 sadece apoptotik nöronların sayısını önemli ölçüde azaltabildi p0.05. CA-074, kaspaz ve katepsin aktivitesini inhibe ederek apoptotik nöronal ölümü azaltmıştır. CA074ün inme hastalarında diğer terapötik ilaçlarla birlikte kullanılması faydalı olabilir.

Makale detayları

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

Yazarlar:

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

Yayıncı:

Eskişehir Osmangazi University

Ana Başlıklar:

Beyin ve Sinir Cerrahisi (Nöroşirurji)

Dergi:

Osmangazi Tıp Dergisi Vol.45 Issue 5, pp.781-790

Belge Tipi:

Makale

Yayın Tarihi:

9/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001293346

pISSN:

1305-4953

ischemia

cerebral

rat model

CA074iskemi

beyin

rat modeli

CA074

Sayı

Makale detayları

Yazarlar: Emre Özkara | Ramazan Durmaz | Zühtü Özbek | Hilmi Özden | Güngör Kanbak | Kubilay Uzuner

Yayıncı: Eskişehir Osmangazi University

Ana Başlıklar: Beyin ve Sinir Cerrahisi (Nöroşirurji)

Dergi: Osmangazi Tıp Dergisi Vol.45 Issue 5, pp.781-790

Belge Tipi: Makale

Yayın Tarihi: 9/2023

Belge Dili: İngilizce

Document ID (Org No.): 2001293346

pISSN: 1305-4953

Sayı

Yazarlar:

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

Yayıncı:

Eskişehir Osmangazi University

Ana Başlıklar:

Beyin ve Sinir Cerrahisi (Nöroşirurji)

Dergi:

Osmangazi Tıp Dergisi Vol.45 Issue 5, pp.781-790

Belge Tipi:

Makale

Yayın Tarihi:

9/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001293346

pISSN:

1305-4953