Doksorubisinin Rat Testis Dokusunda Meydana Getirdiği Hasarların Belirlenmesi ve Çeşitli Antioksidanların Koruyucu Etkileri
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Dosyalar
Tarih
2021
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Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Sunulan tez çalışmasında, beşeri hekimlikte kemoterapötik olarak kullanılan doksorubisinin
rat testis dokusu üzerine zararlı etkileri ve bu zararlı etkiler üzerine timokinon ve resveratrolün
koruyuculuğu araştırıldı.
Çalışmada 10 grup yer aldı. Her bir grupta 8 erkek rat olmak üzere toplamda 80 erkek Albino
Wistar rat kullanıldı. Çalışma grupları kontrol, 15 mg/kg doksorubisin (D), 5mg/kg timokinon (T5),
20mg/kg timokinon (T20), 5mg/kg resveratrol (R5), 20mg/kg resveratrol (R20), D+T5, D+T20,
D+R5 ve D+R20 olarak belirlendi. Kontrol grubuna oral gavaj yöntemiyle gün aşırı serum fizyolojik,
D grubuna 10. gün 15 mg/kg dozda intraperitoneal (İP) olarak doksorubisin, timokinon ve resveratrol
içeren gruplarda ise antioksidanlar oral gavaj yöntemi ile 21 gün boyunca gün aşırı uygulandı.
Yirmibir günün sonunda ratlar anesteziye alınarak servikal dislokasyon yöntemiyle ötenazi edildi.
Spermatolojik ve patolojik parametrelerin ölçülmesi için örnekler alındı. Yapılan ölçümler sonucunda
motilite değerleri karşılaştırıldığında kontrol grubunda motilite %65,83 çıkarken, doksorubisin
uygulanan grupta bu oran %37,5’a kadar düşüş gösterdi (p<0,05). Yine motilite üzerinde, D+T5
(%31,66) ve D+T20 (%40) grupları doksorubisinin zararlı etkisini tersine çeviremezken (p>0,05),
D+R5 (%54,16) ve D+R20 (%61,66) grupları doksorubisin grubundan daha iyi sonuçlar vermeyi
başardı. Epididimal spermatozoa yoğunluğu göz önüne alındığında doksorubisin grubu (66,66x106
)
kontrol grubuna (181,66x106
) oranla spermatozoa yoğunluğunu önemli ölçüde azalttı. Kombine
uygulamalarda ise tüm gruplar doksorubisin grubuna oranla daha iyi sonuçlar vererek spermatozoa
yoğunluğunun düşmesini önledi. Anormal spermatozoa oranları değerlendirildiğinde, doksorubisin
uygulaması anormal spermatozoa oranını (%5,16) diğer gruplara göre önemli derecede artırdı. D+T5
(%2,5), D+T20 (%2,16) ve D+R5 (%1,41) grupları doksorubisin grubuna oranla anormal
spermatozoon oranlarını düşürmeyi başardı. Doksorubisin uygulaması (%50,58) spermatozoa
membran bütünlüğünde önemli oranda hasar oluşturdu. Kombine uygulamalar, spermatozoon
membranında oluşan hasarı kontrol grubu seviyesine getirmede başarısız olmakla birlikte,
doksorubisin’in oluşturduğu hasarı önemli ölçüde azalttı. Mitokondrial aktivite sonuçları
değerlendirildiğinde, doksorubisin grubu (%46,66), diğer gruplara göre önemli oranda düşüşe neden
oldu. D+R5 (%74,1) ve D+R20 (%78,61) gruplarında oluşan hasar doksorubisin grubuna oranla
düzeldi. Doksorubisin (%76,05) uygulaması akrozomal bütünlüğünü kontrol grubuna (%86,41)
oranla önemli oranda düşürdü (p<0,05). Kombine gruplar doksorubisinin oluşturduğu akrozomal
hasarı gidermede başarılı olamadı (p>0,05)
Relatif testis ağırlıkları değerlendirildiğinde, doksorubisin içeren grup (%0,3672) kontrol
grubuna (%0,4371) göre önemli oranda düşüşe neden oldu. D+T5 (%0,4559), D+T20 (%0,4972),
D+R5 (%0,4497) ve D+R20 (0,4845 g) doksorubisinin neden olduğu testis ağırlık kaybını önlemeyi
başardı. Doksorubisin uygulanan hayvanlarda testis germinatif hücre katman kalınlığı (32,9 µm),
kontrol (59,91 µm) grubuna göre oldukça düşük çıktı. Kombine gruplarda ise en başarılı sonucu D+R5 grubu (58,85 µm) verdi. Tubulus seminiferus kontortus çapları dikkate alındığında, kontrol
grubunun ortalama tubulus çapı 308,6 µm iken, doksorubisin uygulanan grupta ortalama 232,33 µm
olarak ölçüldü. D+T5 ve D+T20 uygulamaları, doksorubisinin hasar verici etkisine karşı bir miktar
koruma sağlasa da tubulus çapı değeri kontrol grubuna yaklaşamadı. D+R5 (292,59 µm) ve D+R20
(292,55 µm) grupları tubulus çapı bakımından doksorubisinin hasarını engelleyici etki göstererek
kontrol grubunun ortalamasına oldukça yaklaştırdı (p<0,05). Johnson’un testiküler skorlaması
incelendiğinde, en iyi sonucu 9,0833 ile kontrol grubu verdi. Yalnızca doksorubisin içeren grup
(4,0833) oldukça düşük bir skor alırken, timokinon (T5 ve T20) ve resveratrol (R5 ve R20) içeren
gruplar kontrol grubuyla benzer sonuçları verdi (p>0,05).
Kontrol grubunda Bax değeri 4,19 olarak gözlenirken, doksorubisin grubunda (12,37)
oldukça yüksek çıktı. Doksorubisin ile kombine gruplarda Bax miktarı doksorubisin grubundan daha
düşük çıksa da en iyi sonucu 5,75 ile D+T5 grubu verdi. Bcl-2 sonuçları değerlendirildiğinde,
doksorubisin grubunun Bcl-2 değeri (2,86) diğer tüm gruplara oranla önemli ölçüde düşük çıktı.
Kontrol grubunda bu değer 17,25 çıkarken, D+T5 (18,57), D+T20 (16,36) ve D+R20 (18,98) kontrol
grubuyla benzer ve D grubundan daha yüksek Bcl-2 skorları verdi (p<0,05).
Sunulan tez çalışmasında, doksorubisinin testis dokusuna zarar verici etkileri ortaya kondu.
Timokinon ve resveratrol bileşikleri ise testiküler dokuda doksorubisine bağlı oluşan hasarı önlemede
hem spermatolojik hem de patolojik açıdan başarılı bulundu.
In the presented thesis, harmful effects of doxorubicin on rat testicular tissue were determined and the protectiveness of thymoquinone and resveratrol have been studied. A total of 80 male Albino Wistar rats, including 10 groups and 8 male rats in each group, were used in the study. Experimental groups were designed as; control, 15 mg/kg doxorubicin (D), 5mg/kg thymoquinone (T5), 20mg/kg thymoquinone (T20), 5mg/kg resveratrol (R5), 20mg/kg resveratrol (R20), D+T5, D+T20, D+R5 and D+R20. Physiological Saline was administered to the control group every other day by oral gavage. On the doxorubicin group, 15 mg/kg doxorubicin injected intraperitoneally on 10th day of 21 day period and same procedures has been applied with control group then administrated by oral gavage every other day for 21 days. At the end of the 21th day, rats euthanised by cervical dislocation method. After the euthanise, samples collected for spermatologic and pathologic measurements. When the motility values were compared, while the motility was 65.83% in the control group, this rate decreased to 37.5% in the doxorubicin applied groups (p<0,05). D+T5 (31.66%) and D+T20 (40%) groups could not reverse the harmful effect of doxorubicin (p>0,05), but the D+R5 (54.16%) and D+R20 (61.66%) groups were better than the doxorubicin group. Considering the concentration of epididymal spermatozoa, the doxorubicin group (66.66x106 ) significantly reduced the spermatozoa concentration compared to the control group (181.66x106 ). In combined applications, all groups provided better results when compared to the doxorubicin group and preventing the decrease in spermatozoa concentration. When the abnormal spermatozoa rates were evaluated, doxorubicin administration significantly increased the rate of abnormal spermatozoa (5.16%) compared to the other groups. Among the combined groups, D+T5 (2,5%), D+T20 (2,16%) and D+R5 (1,41%) groups were able to reduce the abnormal spermatozoon rates when compared to the doxorubicin group. In terms of spermatozoon membrane integrity, doxorubicin administration (50.58%) caused significant damage to membrane integrity. Although the combined applications failed to bring the damage to the spermatozoon membrane to the level of the control group, it significantly reduced the damage caused by doxorubicin. When the mitochondrial activity results were evaluated, the doxorubicin group (46.66%) caused a significant decrease compared to the other groups. Damages on the D+R5 (74.1%) and D+R20 (78.61%) groups were ameliorated according to the doxorubicine group. In acrosomal integrity, administration of doxorubicin (76.05%) significantly reduced acrosomal integrity compared to the control group (86.41%, p<0,05). Combined groups could not succeed in removing the acrosome damage consisting of doxorubicin (p>0,05). When the relative testicular weights were evaluated, the group containing doxorubicin (0.3672%) caused a significant decrease compared to the control group (0.4371%), D+T5 (0.4559%), D+T20 (0.4972%), D+R5 (0.4497%) and D+R20 (0.4845 g) prevented testicular weight loss caused by doxorubicin. Germinative cell layer thickness in animals treated with doxorubicine (32.9 µm) decreased considerably compared to the control (59.91 µm). In the combined groups, D+R5 (58.85 µm) gave the most successful result. When tubulus seminiferus contortus diameters were taken into consideration, it was noted that while the avarege tubulus diameter of control was 308.6 µm, it was measured as 232.33 µm in doxorubicine group. Although D+T5 and D+T20 groups provided some protection against the damaging effect of doxorubicin, their tubulus diameters could not come close to the control. D + R5 (292.59 µm) and D+R20 (292.55 µm) groups preventing the damaging effect of doxorubicin in terms of tubular diameter and brought it closer to the average value of the control (p<0,05). When Johnson's testicular score was examined, the best result was found in the control (9.0833). While the only doxorubicin (4.0833) group had considerably low score, the groups containing thymoquinone (T5 and T20) and resveratrol (R5 and R20) gave similar results to that of control (p>0,05). While Bax was 4.19 in the control, this value was 12.37 in the doxorubicin group. Although the amount of Bax in the groups combined with doxorubicin was lower than the doxorubicin group, the D + T5 group (5.75) gave the best result. When the Bcl-2 results were evaluated, the Bcl-2 value of the doxorubicin group (2.86) was significantly lower than all other groups. While this value was 17.25 in the control group, D + T5 (18.57), D + T20 (16.36) and D + R20 (18.98) gave similar results to that of control (p>0,05) and gave higher Bcl-2 scores than doxorubicin (p<0,05). In the presented thesis, the damaging effects of doxorubicin on testicular tissue were revealed. Thymoquinone and resveratrol compounds were found to be both spermatologically and pathologically successful in preventing doxorubicin-induced damage to testicular tissue.
In the presented thesis, harmful effects of doxorubicin on rat testicular tissue were determined and the protectiveness of thymoquinone and resveratrol have been studied. A total of 80 male Albino Wistar rats, including 10 groups and 8 male rats in each group, were used in the study. Experimental groups were designed as; control, 15 mg/kg doxorubicin (D), 5mg/kg thymoquinone (T5), 20mg/kg thymoquinone (T20), 5mg/kg resveratrol (R5), 20mg/kg resveratrol (R20), D+T5, D+T20, D+R5 and D+R20. Physiological Saline was administered to the control group every other day by oral gavage. On the doxorubicin group, 15 mg/kg doxorubicin injected intraperitoneally on 10th day of 21 day period and same procedures has been applied with control group then administrated by oral gavage every other day for 21 days. At the end of the 21th day, rats euthanised by cervical dislocation method. After the euthanise, samples collected for spermatologic and pathologic measurements. When the motility values were compared, while the motility was 65.83% in the control group, this rate decreased to 37.5% in the doxorubicin applied groups (p<0,05). D+T5 (31.66%) and D+T20 (40%) groups could not reverse the harmful effect of doxorubicin (p>0,05), but the D+R5 (54.16%) and D+R20 (61.66%) groups were better than the doxorubicin group. Considering the concentration of epididymal spermatozoa, the doxorubicin group (66.66x106 ) significantly reduced the spermatozoa concentration compared to the control group (181.66x106 ). In combined applications, all groups provided better results when compared to the doxorubicin group and preventing the decrease in spermatozoa concentration. When the abnormal spermatozoa rates were evaluated, doxorubicin administration significantly increased the rate of abnormal spermatozoa (5.16%) compared to the other groups. Among the combined groups, D+T5 (2,5%), D+T20 (2,16%) and D+R5 (1,41%) groups were able to reduce the abnormal spermatozoon rates when compared to the doxorubicin group. In terms of spermatozoon membrane integrity, doxorubicin administration (50.58%) caused significant damage to membrane integrity. Although the combined applications failed to bring the damage to the spermatozoon membrane to the level of the control group, it significantly reduced the damage caused by doxorubicin. When the mitochondrial activity results were evaluated, the doxorubicin group (46.66%) caused a significant decrease compared to the other groups. Damages on the D+R5 (74.1%) and D+R20 (78.61%) groups were ameliorated according to the doxorubicine group. In acrosomal integrity, administration of doxorubicin (76.05%) significantly reduced acrosomal integrity compared to the control group (86.41%, p<0,05). Combined groups could not succeed in removing the acrosome damage consisting of doxorubicin (p>0,05). When the relative testicular weights were evaluated, the group containing doxorubicin (0.3672%) caused a significant decrease compared to the control group (0.4371%), D+T5 (0.4559%), D+T20 (0.4972%), D+R5 (0.4497%) and D+R20 (0.4845 g) prevented testicular weight loss caused by doxorubicin. Germinative cell layer thickness in animals treated with doxorubicine (32.9 µm) decreased considerably compared to the control (59.91 µm). In the combined groups, D+R5 (58.85 µm) gave the most successful result. When tubulus seminiferus contortus diameters were taken into consideration, it was noted that while the avarege tubulus diameter of control was 308.6 µm, it was measured as 232.33 µm in doxorubicine group. Although D+T5 and D+T20 groups provided some protection against the damaging effect of doxorubicin, their tubulus diameters could not come close to the control. D + R5 (292.59 µm) and D+R20 (292.55 µm) groups preventing the damaging effect of doxorubicin in terms of tubular diameter and brought it closer to the average value of the control (p<0,05). When Johnson's testicular score was examined, the best result was found in the control (9.0833). While the only doxorubicin (4.0833) group had considerably low score, the groups containing thymoquinone (T5 and T20) and resveratrol (R5 and R20) gave similar results to that of control (p>0,05). While Bax was 4.19 in the control, this value was 12.37 in the doxorubicin group. Although the amount of Bax in the groups combined with doxorubicin was lower than the doxorubicin group, the D + T5 group (5.75) gave the best result. When the Bcl-2 results were evaluated, the Bcl-2 value of the doxorubicin group (2.86) was significantly lower than all other groups. While this value was 17.25 in the control group, D + T5 (18.57), D + T20 (16.36) and D + R20 (18.98) gave similar results to that of control (p>0,05) and gave higher Bcl-2 scores than doxorubicin (p<0,05). In the presented thesis, the damaging effects of doxorubicin on testicular tissue were revealed. Thymoquinone and resveratrol compounds were found to be both spermatologically and pathologically successful in preventing doxorubicin-induced damage to testicular tissue.
Açıklama
Anahtar Kelimeler
Doksorubisin, timokinon, resveratrol, rat, testis, Doxorubicin, thymoquinone, testes
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Öztürk, A. E., (2021). Doksorubisinin Rat Testis Dokusunda Meydana Getirdiği Hasarların Belirlenmesi ve Çeşitli Antioksidanların Koruyucu Etkileri. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimler Enstitüsü, Konya.
