The effect of vitamin D deficiency and 1,25(OH)2D3 treatment on oxidative stress and Nrf2-antioxidant signaling in ethanol-induced hepatotoxicity

Abstract

Aim: Vitamin D deficiency (VDD) is suggested to enhance hepatotoxicity in chronic liver diseases. However,
there is limited knowledge about the association between VDD and alcoholic liver damage. Therefore, the effect
of VDD on ethanol (EtOH)-induced hepatotoxicity was investigated in this study. Moreover, the role of the
Nrf2-antioxidant signaling pathway in the hepatoprotective potential of 1,25(OH)2D3 was also searched in
EtOH-treated rats.
Methods: Male Wistar rats were fed on VDD-diet for 12 weeks. EtOH (5-20%) was applied in drinking water in
increasing concentrations for the last 8 weeks. In addition, one group of rats were injected with 1,25(OH)2D3
(5μg/kg; twice a week; i.p.) during this period. Hepatic triglyceride and hydroxyproline levels, inflammation
markers, lipid peroxides, protein carbonyls, mRNA expressions of Nrf2, superoxide dismutase (SOD), and
glutathione peroxidase (GSH-Px), SOD and GSH-Px activities, glutathione levels and histopathology were
examined.
Results: EtOH application caused steatosis and fibrosis, elevated hepatic TG, lipid peroxide, protein carbonyls
and hydroxyproline levels and inflammation markers. VDD did not aggravate EtOH-induced liver damage,
steatosis and inflammation, but reactive oxygen species and lipid peroxide levels were slightly increased in
VDD+EtOH group. Gene expressions of Nrf2-SOD-GSH-Px, enzyme activities and glutathione levels were also
higher in VDD+EtOH group than EtOH group. Additionally, 1,25(OH)2D3 elevated mRNA expressions and
activities of SOD and GSH-Px in EtOH-treated rats.
Conclusion: Our results indicate that VDD diet did not cause an additive effect on EtOH-induced hepatotoxicity.
Moreover, it was detected that the activation of Nrf2-antioxidant signaling pathway may play a role in the
protective effect of 1,25(OH)2D3 against EtOH-induced hepatotoxicity

Keywords

• Vitamin D deficiency
• 1,25(OH)2D3
• Ethanol
• Liver damage
• Oxidative stress
• Nrf2

D vitamini eksikliği ve 1,25(OH)2D3 uygulamasının etanole bağlı karaciğer hasarında oksidatif stres ve Nrf2-antioksidan sinyal sistemi üzerine etkisi

Öz

Amaç: D vitamini eksikliğinin (VDE) kronik karaciğer hastalıklarında karaciğer hasarını arttırdığı ileri
sürülmektedir. Bununla birlikte, VDE ile alkole bağlı karaciğer hasarı arasındaki ilişkiye ait bilgiler sınırlıdır.
Bu nedenle, bu çalışmada VDE'nin etanol (EtOH) ile indüklenen karaciğer hasarı üzerindeki etkisi araştırıldı.
Ayrıca, EtOH uygulanan sıçanlarda 1,25(OH)2D3'ün karaciğeri koruyucu potansiyelinde Nrf2-antioksidan sinyal
yolunun rolü de araştırıldı.
Yöntemler: Erkek Wistar sıçanlar 12 hafta süreyle VDE diyet ile beslendi. Son 8 hafta içme suyunda artan
konsantrasyonlarda EtOH (% 5-20) verildi. Ayrıca, bir grup sıçana bu süreçte 11,25(OH)2D3 (5μg/kg; haftada iki
kez; i.p.) uygulandı. Karaciğerde trigliserit (TG) ve hidroksiprolin düzeyleri, inflamasyon göstergeleri, lipid
peroksitler, protein karboniller, Nrf2, süperoksit dismutaz (SOD) ve glutatyon peroksidaz (GSH-Px)’ın mRNA
ekspresyonları, SOD ve GSH-Px aktiviteleri, glutatyon düzeyleri ve histopatolojik incelemeler yapıldı.
Bulgular: EtOH uygulaması steatoz ve fibroza, karaciğerde TG, lipit peroksit, protein karbonil ve hidroksiprolin
düzeyleri ile inflamasyon göstergelerinde artışa neden oldu. VDE, EtOH’a bağlı karaciğer hasarını, steatoz ve
fibrozu yoğunlaştırmadı. Ancak, reaktif oksijen türleri ve lipit peroksit düzeylerinde VDE+EtOH grubunda
ılımlı artış bulundu. Nrf2-SOD-GSH-Px’in gen ekspresyonları, enzim aktiviteleri ve glutatyon düzeyleri de
VDE+EtOH grubunda EtOH grubundan daha yüksekti. Ayrıca, 1,25(OH)2D3, EtOH uygulanan sıçanlarda
karaciğerde SOD ve GSH-Px’ın mRNA ekspresyonlarını ve aktivitelerini arttırdı.
Sonuç: Bulgularımız VDE diyetin EtOH ile indüklenen karaciğer hasarı üzerinde ek bir etki oluşturmadığını
gösterdi. Ayrıca EtOH’a bağlı karaciğer hasarında 11,25(OH)2D3'ün koruyucu etkisinde Nrf2-antioksidan sinyal
yolunun aktivasyonunun rol oynayabileceği saptandı. 

Anahtar Kelimeler

• D vitamini eksikliği
• 1,25(OH)2D3
• Etanol
• Karaciğer hasarı
• Oksidatif stres
• Nrf2

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