Emin KAYMAK, Derya KARABULUT, Emel ÖZTÜRK, Ali Tuğrul AKIN, Nurhan KULOĞLU, Birkan YAKAN

B12 vitamini sıçanlarda metotreksatın neden olduğu akciğer hasarını azaltır: Histopatolojik, immünohistokimyasal ve biyokimyasal bir çalışma

Amaç: Metotreksat (MTX) önemli bir kemoterapötik olup kanser hastalarında kullanılan önemli bir antikanserojen olmakla birlikte diğer dokularda yan etkilere yol açmaktadır. Bu yan etkiler arasında akciğer toksisitesi de yer almaktadır. B12 Vitamini güçlü bir antioksidandır ve reaktif oksijen türlerinin azalmasını sağlar. Bu çalışma, B12 vitamininin sıçan akciğerinde metotreksat kaynaklı hasara karşı koruma sağlayıp sağlayamayacağını belirlemek için tasarlanmıştır. Yöntem: Toplam 32 erkek Wistar albino sıçanı dört gruba ayrılmıştır: Kontrol grubuna (n=8) deney boyunca intraperitoneal (i.p.) salin uygulanmıştır. B12 vitamini (B12) grubuna (n=8) 3 μg/kg/i.p. 14 gün boyunca B12 vitamini verilmiştir. Metotreksat (MTX) grubuna 20 mg/ kg/i.p. tek doz (MTX) enjeksiyonu deneyin sekizinci gününde yapılmıştır. MTX+B12 grubuna deneyin sekizinci gününde MTX tek doz 20 mg/kg/ i.p. + 3 μg/kg/i.p. B12 vitamini günlük deney boyunca uygulandı. Elde edilen akciğer dokularına histopatolojik, immünohistokimyasal ve biyokimyasal yöntemler uygulanmıştır. Akciğer dokuları Masson’s Trikrom (MT) ile boyanmıştır. Ayrıca α-Sma, Laminin, PCNA ve TNF-α antikorları immünohistokimya ile boyandı. Akciğer doku homojenatlarında ise süperoksit dismutaz (SOD), katalaz (CAT) ve malondialdehit (MDA) seviyeleri değerlendirilmiştir. Bulgular: Metotreksat, MDA ve IL-6 seviyelerinde ve α-Sma, Laminin ve TNF-α ekspresyonunda ve akciğer dokusunda apoptotik hücre sayısında artışa neden olmuştur. Ayrıca MTX grubunda PCNA ekspresyonu ve SOD ve CAT seviyelerinde azalma görülmüştür. B12 vitamini, MDA ve IL-6 düzeylerindeki artışı ve α-Sma, Laminin ve TNF-α ekspresyonunu engellemiştir. B12 vitamininin antioksidan kapasiteyi artırdığı tespit edilmiştir. Sonuç: B12 vitamininin metotreksat kaynaklı akciğer toksisitesinde oksidatif stress, inflamasyon, fibrosis ve apoptosis gibi faktörler üzerinde etkili olduğu ve hasarı azalttığı görülmüştür. Bu çalışma, metotreksatın neden olduğu akciğer hasarı için B12 vitamini kullanımının faydalı olacağını göstermiştir. Kanser ilacı kullanımında yan etkileri azaltmada B12 vitamininin gözardı edilmemesi gerektiğini düşündürmektedir.

Vitamin B12 alleviates methotrexate-induced lung injury in rat: A histopathological, immunohistochemical and biochemical study

Objective: Methotrexate (MTX) is an important chemotherapeutic and an important anticarcinogen used in cancer patients, but it causes side effects in other tissues. Among these side effects is lung toxicity. Vitamin B12 is a powerful antioxidant and reduces reactive oxygen species. This study was designed to determine whether vitamin B12 could protect against methotrexate-induced damage in rat lungs. Methods: A total of 32 male Wistar albino rats were divided into four groups: The control group (n=8) received intraperitoneal (i.p.) saline throughout the experiment. Vitamin B12 group (B12) (n=8) 3 μg/ kg/i.p. Vitamin B12 was administered for 14 days. The methotrexate (MTX) group received a single dose MTX injection at 20 mg/kg/i.p. on the 8th day of the experiment. On the 8th day of the experiment, a single dose of MTX 20 mg/kg/i.p. was given to the MTX+B12 group. + 3 μg/kg/i.p. Vitamin B12 was administered daily throughout the experiment. Histopathological, immunohistochemical, and biochemical methods were applied to the obtained lung tissues. Lung tissues were stained with Masson’s Trichrome (MT). In addition, α-Sma, Laminin, PCNA, and TNF-α antibodies were stained by immunohistochemistry. Superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) levels were evaluated in lung tissue homogenates. Results: MTX caused an increase in MDA and IL-6 levels and expression of α-Sma, Laminin, and TNF-α, and the number of apoptotic cells in lung tissue. It also caused a decrease in PCNA expression and SOD and CAT levels in the MTX group. Vitamin B12 inhibited the increase in MDA and IL-6 levels and the expression of α-Sma, Laminin, and TNF-α. Vitamin B12 was found to increase antioxidant capacity. Conclusion: Vitamin B12 has been shown to be effective on factors such as oxidative stress, inflammation, fibrosis, and apoptosis in MTX-induced lung toxicity and reduce damage. We observed that MTX stimulated lung injury decreased with vitamin B12 treatment It suggests that vitamin B12 should not be ignored in reducing side effects in cancer drug use.

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