SERULEİNLE OLUŞTURULMUŞ SIÇAN AKUT PANKREATİT MODELINDE BETA GLUKAN VE FUKOKSANTİNİN ETKİSİ The Effect of Beta Glucan and Fucoxanthine in a Serulein- Induced Acute Pancreatitis in Rats

Öz

ÖZET Amaç: Bu çalışmada, Beta Glukan, fukoksantin ve kombinasyonlarının serulein kaynaklı akut pankreatit (AP) sıçan modelindeki etkileri araştırıldı. Gereç ve Yöntem: Sıçanlar kontrol ve AP gruplarına ayrıldı. AP bir saat arayla dört kez serulein (20 ug/kg/ ip) enjeksiyonuyla oluşturuldu. Serulein enjeksiyonundan 3 gün önce AP gruplarına günde 1 kez taşıyıcı, Beta Glukan (BG;40 mg/kg/po), fukoksantin (Fuko;40 mg/kg/po) veya Beta Glukan (BG;40 mg/kg/po) +fukoksantin (Fuko;40 mg/kg/po) uygulandı. Son enjeksiyondan sonraki 8. saatte ötenazi gerçekleştirildi. Serumda amilaz ve lipaz, pankreas dokusunda interlökin-1β (IL-1β), interlökin-6 (IL-6), interlökin-10 (IL- 10), lucigenin kemilüminesans (CL), malondialdehit (MDA), miyeloperoksidaz (MPO) enzim aktivitesi ve Hematoksilen&Eosin (H&E), Siklooksijenaz-2 (COX-2), nükleer faktör kappa B (NF-κB) histopatolojik incelemeleri yapıldı. İstatistiksel analizler için tek yönlü ANOVA ve Bonferroni post-hoc testi uygulandı. Bulgular: Taşıyıcı grubunda; amilaz, lipaz, lusigenin, IL-1β ve IL-6 düzeyleri kontrole göre (p <0,01-0,0001) yükselirken, GSH ve IL-10 düzeyleri azaldı (p<0,0001). Lipaz ve amilaz düzeyleri; BG, Fuko ve BG+Fuko tedavileri ile taşıyıcıya göre azaldı (p<0,05-0,001). IL-6 düzeyleri, Fuko ve BG+Fuko grubunda taşıyıcıya göre azaldı (p <0,01). IL-1β düzeyleri; BG, Fuko ve BG+Fuko gruplarında taşıyıcı grubuna göre azaldı (p<0,001- 0.0001). IL-10 düzeyi yalnızca BG grubunda taşıyıcı grubuna göre arttı (p<0,01). H&E, COX-2 skorları taşıyıcı grubunda kontrole göre artarken (p<0,0001); BG, Fuko ve BG+Fuko tedavileri ile azalma gözlendi (p<0,05- 0,001). Taşıyıcı grubunda artan NF-κB skorları (p<0,0001), BG tedavisi ile azaldı (p<0,01). Taşıyıcı grubundaki artan lusigenin kemiluminesans, MPO ve MDA düzeyleri (p <0,01-0,0001) tüm tedavilerle azaldı (p <0,01- 0,0001). Sonuç: Beta Glukan ve fukoksantin tedavisi pro-inflamatuvar sitokinleri, COX-2 düzeyini ve oksidatif belirteçleri azaltarak anti-inflamatuvar sitokin düzeyini arttırarak AP gelişimini hafifletmiştir. Anahtar sözcükler: Beta glukan; fukoksantin; COX-2; oksidatif stres ABSTRACT Aim: This study aims to investigate the effects of Beta Glucan, fucoxanthin and their combinations in acute pancreatitis (AP) rat model. Material and Methods: Rats were divided into control and AP groups. The AP was induced by injection of cerulein (20 μg/kg/ip) four times with an hour interval. Vehicle, Beta Glucan (BG;40 mg/kg/po), fucoxanthin (Fuco;40 mg/kg/po) or Beta Glucan (BG;40 mg/kg/+ fucoxanthin (Fuco;40 mg/kg/po) were administered 3 days before cerulein injection. Euthanasia was performed at the 8th hour after the last injection. Serum and pancreatic tissue collected for biochemical and histopathological examinations. One-way ANOVA and Bonferroni post-hoc test was used for statistical analysis. Results: Amylase, lipase, lucigenin, interleukin-1β, and interleukin-6 levels are significantly elevated in the vehicle-treated AP groups (p<0.01-0.0001) while IL-10 was decreased (p<0.0001). Lipase and amylase levels decreased in the BG, Fuco and BG+Fuco groups compared to vehicle group (p<0.05-0.001). IL-6 levels, decreased in the fuco and BG+Fuco group (p <0.01). IL-1β levels decreased in BG, Fuco and BG+Fuco groups compared to vehicle group (p<0,001-0.0001). IL-10 levels increased in BG group compared to the vehicle group (p<0.01). H&E and COX-2 scores increased in the vehicle groups, compared to the control group (p<0.0001). All treatments decreased that scores (p<0.05-0.001). Increased NF-κB scores with vehicle (p<0.0001), decreased with BG treatment (p<0.01). Increased lucigenin, MPO and MDA levels (p<0.01- 0.0001), decreased with BG, Fuco and BG+Fuco treatments (p<0.01-0.0001). Conclusion: Treatment of Beta Glucan and fucoxanthin alleviated AP development by decreasing proinflammatory cytokines, COX-2 and oxidative markers and increasing anti-inflammatory cytokine. Keywords: Beta glucan; fucoxanthine; COX-2; Oxidative stress

Anahtar Kelimeler

• SERULEİNLE OLUŞTURULMUŞ SIÇAN AKUT PANKREATİT MODELINDE BETA GLUKAN VE FUKOKSANTİNİN ETKİSİ The Effect of Beta Glucan and Fucoxanthine in a Serulein- Induced Acute Pancreatitis in Rats

Kaynakça

1. 1. Su KH, Cuthbertson C, Christophi C. Review of experimental animal models of acute pancreatitis. HPB (Oxford). 2006; 8(4):264–286. doi:10.1080/13651820500467358 2. Escobar J, Pereda J, Arduini A, Sandoval J, Sabater L, Aparisi L, et al. Cross-Talk between Oxidative Stress and Pro-Inflammatory Cytokines in Acute Pancreatitis: A Key Role for Protein Phosphatases. Current Pharmaceutical Design. 2009; 15(26), 3027–3042. doi:10.2174/138161209789058075 3. Lin ZS, Ku CF, Guan YF, Xiao HT, Shi XK, Wang HQ, et al. Dihydro-Resveratrol Ameliorates Lung Injury in Rats with Cerulein-Induced Acute Pancreatitis. Phytother Res. 2016 ;30(4):663-70. doi: 10.1002/ptr.5576. 4. Rakonczay Jr Z, Hegyi P, Taka´cs T, McCarroll J, Saluja AK. The role of NF-kB activation in the pathogenesis of acute pancreatitis. Gut. 2008; 57:259–267. doi:10.1136/gut.2007.124115 5. Xiong J, Wang K, Yuan C, Xing R, Ni J, Hu G, et al. Luteolin protects mice from severe acute pancreatitis by exerting HO-1-mediated anti-inflammatory and antioxidant effects. Int J Mol Med. 2017; 39(1): 113–125. 6. Altavilla D, Famulari C, Passaniti M, Campo GM, Macrì A, Seminara P, et al. Lipid peroxidation inhibition reduces NF-kappaB activation and attenuates cerulein-induced pancreatitis. Free Radic Res. 2003; 37:425–35. 7. de Almeida JL, Jukemura J, Coelho AM, Patzina RA, Machado MC, da Cunha JE. Inhibition of cyclooxygenase-2 in experimental severe acute pancreatitis. Clinics (Sao Paulo). 2006;61(4):301-6. doi:10.1590/s1807-59322006000400005 8. Song AM, Bhagat L, Singh VP, Van Acker GGD, Steer ML, Saluja AK. Inhibition of cyclooxygenase-2 ameliorates the severity of pancreatitis and associated lung injury. Am J Physiol Gastrointest Liver Physiol. 2002; 283:1166-74. 9. Raposo MF, de Morais AM, de Morais RM. Carotenoids from Marine Microalgae: A Valuable Natural Source for the Prevention of Chronic Diseases. Mar Drugs. 2015; 13(8):5128–5155. 10. Heo SJ, Ko SC, Kang SM, Kang HS, Kim JP, Kim SH, et al. Cytoprotective effect of fucoxanthin isolated from brown algae Sargassum siliquastrum against H2O2-induced cell damage. Eur. Food Res. Technol. 2008; 228:145–151. doi: 10.1007/s00217-008-0918-7.