Annona Muricata Yaprak Ekstratının Spinal Korddaki Radyoprotektif Etkileri ve Histopatolojik Sonuçları

Öz

Amaç: Radyoterapi (RT), primer ve metastatik tümörler için önemli bir kanser tedavisidir. Çok sayıda çalışma, DNA'nın, l ipitlerin ve proteinlerin, iyonize radyasyon (IR) tarafından indüklenen serbest radikaller tarafından saldırıya uğradığını ve dolayısıyla ciddi hücresel hasara yol açtığını göstermiştir. Yaygın olarak "graviola" olarak bilinen Annona muricata (AM) bitkisi tropik iklime sahip ülkelerde (Güney Amerika, Batı Afrika, Güneydoğu Asya) yetişmekte olup antioksidan, antikanser, analjezik ve antispazmodik ajan olarak kullanılmaktadır. Yerel halk tarafından geleneksel ve alternatif tıp amaçlı yaprakları, kökü, kabuğu, tohumları ve meyvesi çok çeşitli formlarda kullanılmaktadır. Çalışmamızda graviola yaprakları kullanılmış olup, Güney Amerika’nın yerel halkından temin edilmiştir. Bu çalışma, graviolanın iyonize radyasyonun omurilik hasarına karşı olası koruyucu etkisini değerlendirmeyi amaçladı. Gereç ve Yöntemler: Benzer fizyolojik özelliklere sahip 32 adet erişkin dişi (250±50gr) Wistar Albino rat randomize olarak seçildi. Ratlar Grup 1 (n=8) (control grubu), Grup 2 (n=8) (Graviola grubu), Grup 3 (n=8) (RT grubu), Grup 4 (n=8) (Graviola+ RT grubu) şeklinde gruplandı. Öz, etanol bazlı filtrasyon yoluyla Graviola bitkisinin kurutulmuş yapraklarından elde edildi. Oluşan öz, ratlara aktif bir bileşen olarak uygulandı. Prone pozisyonda sabitlenmiş ratların spinal korduna toplam 20Gy (1GY/dk) dozda tek bir fraksiyon halinde uygulandı. Tüm analizler SPSS ile yapıldı. Sonuçlar %95 güven aralığında değerlendirilecek olup, p<0.05 değeri anlamlı kabul edilecektir. Bulgular: Dört ayrı grupta inflamatuar hücre infiltrasyonu, ödem, nöron şişmesi, gliosit hiperplazisi, nekroz, spinal kord kavitasyonu, Nissl cisimcik kaybı, çekirdek çevresinde kavitasyon, piknotik çekirdek gibi parametreler incelendi. İstatistiksel analiz dört grup arasında yukarıdaki parametreler değerlendirilerek yapılmıştır. Doku ve hücrelerin histomorfolojik bulguları skorlandı. Her kriter 0 ile 3 arasında puanlandı. (0=normal, 1=Hafif, 2=orta ve 3=ağır). Sonuç: Ratların RT'den önce graviola ile tedavisinin, control grubuna kıyasla omurilikte nekrotik hasarları önlediğini göstermiştir. RT'den önce graviola yaprak ekstresi ile yapılan ön tedavinin, graviolanın antiinflamatuar özelliğinin nöronal korumaya katkıda bulunduğunu desteklemektedir.

Anahtar Kelimeler

• Annona muricata
• spinal kord
• iyonize radyasyon
• radyoprotektif

Etik Beyan

Etik Kurul Tarih: 02.07.2020 Etik kurul No: 2020-18-02/07 Sıçanlarda İyonize Radyasyona Bağlı Spinal Kord Hasarının Önlenmesinde Annona Muricata (Graviola)'nın Etkisi Protective Effects of Annona Muricata (Gravıola) on Radıatıon Spınal Cord

Radioprotective Effects of Annona Muricata Leaf Extract in the Spinal Cord

Öz

Objective: The spinal cord is often involved in radiotherapy to treat malignancies and is considered as one of the most critical dose-limiting organs. Histopathological changes of radiation-induced spinal cord injury include gliosis, demyelination, and necrosis in the white matter, as well as vascular changes in both white and gray matter. All aerial parts of Annona muricata (AM) are used in traditional medicine practices, and previous studies revealed that AM leaf extract has radioprotective effect against gamma- irradiation. This study aimed to evaluate the possible protective effect of AM leaf extract against ionizing radiation spinal cord injury. Methods: A total of 37 adult female Wistar Albino rats were divided into 4 groups (1 control and 3 intervention groups): Group 1 (control) received 0.01 mL/kg of distilled water by oral gavage once a day for 7 days, group 2 (AM group) received 300 mg/kg of AM leaf extract by oral gavage once a day for 7 days, group 3 (IR group) received a single dose of spinal cord radiation (20 Gy) after a 7-day treatment with 0.01 mL/kg of physiologic serum (saline), and group 4 (AM + IR group) received 300 mg/kg of AM leaf extract by oral gavage once a day for 7 days, with spinal cord radiation (20 Gy) applied 1 hour after the last gavage. After the sacrificing, spinal cords were dissected out, embedded in paraffin, and stained with Hematoxylin-Eosin, and 11 parameters were microscopically examined and scored. Results: AM administration prior to irradiation (IR) lowered the histopathological scores for all parameters that were verified in the IR group at most severe levels: inflammatory cells infiltration, edema, neuron swelling, gliocytes hyperplasia, spinal cord cavitation (loosened tissue structure), pyknotic nuclei, vacuolar denaturation, and loss of Nissls substance. Necrosis was present in IR group and was higher compared to Control and AM treated groups. We noticed that necrosis was present in the AM group too and was higher compared to the control. Conclusion: The data of our study suggest that pretreatment with 300 mg/kg of AM leaf extract prior to IR reduce inflammatory cells infiltration, edema, and neuron swelling. This supports the fact that AM-proposed anti-inflammatory property may contribute to neuroprotection. Similarly, AM was documented to have efficient protection on neural cells from apoptosis or necrosis.

Anahtar Kelimeler

• Annona muricata
• Spinal cord
• Irridation
• Radioprotective

Kaynakça

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