Ratlarda Deneysel Olarak Oluşturulan Kolit Modelinde N-Asetil Sistein ve β-Glukanın Etkileri

Yıl 2021, Cilt: 16 Sayı: 3, 357 - 364, 01.11.2021

Murat İspiroğlu , Bülent Kantarçeken , Harun Çıralık , Ertan Bulbuloglu

https://doi.org/10.17517/ksutfd.817235

Öz

Giriş-Amaç
Ülseratif kolit, kolonu diffüz tutan mukozal inflamasyonla karakterize rekürren, idiyopatik ve kronik bir hastalıktır. Kolitde, oksidan/antioksidan dengenin bozulduğu gözlenmiştir. N-asetilsistein ve β-glukan ise antioksidan, anti-inflamatuvar özellikte olan maddeler olup kolitde yararlı etkilerini araştırmayı amaçladık.
Materyal-Metod
220-250 gr 50 adet erkek Wistar Albino rat kullanıldı.
Grup I (Kontrol):Tek doz rektal saline uygulandı, sonrası 6 gün normal besin verildi.
Grup II (Kolit):Tek doz rektal asetik asit uygulandı, sonrası 6 gün normal besin verildi.
Grup III (β-Glukan+Kolit):Tek doz oral 100mg/kg β-glukan verildikten 1 saat sonra rektal asetik asit uygulandı. Sonraki 6 gün oral 100/mg/kg/gün β-glukan verildi.
Grup IV (N-asetilsistein+Kolit):Tek doz oral 200mg/kg N-asetilsistein verildikten 1 saat sonra rektal asetik asit uygulandı. Sonraki 6 gün oral 200mg/kg/gün N-asetilsistein verildi.
Grup V (N-asetilsistein+β-glukan+Kolit): Tek doz oral 200mg/kg N-asetilsistein+100 mg/kg β-glukan verildi. 1 saat sonra rektal asetik asit uygulandı. Sonraki 6 gün oral 100 mg/kg/gün β-glukan + 200 mg/kg/gün N-asetilsistein verildi.
Çalışmanın sonunda kalın barsak distal 8 cm’lik kısmı çıkarıldı. Histopatolojik ve biyokimyasal inceleme için örnekler alındı.
Bulgular
Tedavi alanlar da almayanlara göre MDA ve MPO düzeyleri anlamlı düşük; SOD, KAT düzeyleri ise anlamlı yüksek olduğu gözlendi. Tedavi grupları arasında, MDA ve MPO düzeylerinde anlamlı fark yoktu. Diğer antioksidan enzimler (SOD, KAT); N-asetilsistein grubuyla β-glukan grubu arasında anlamlı fark yok iken kombinasyon verilen grupta antioksidan savunma, N-asetilsistein grubundan anlamlı olarak düşük gözlendi. Histopatolojik skor ortalamaları değerlendirildiğinde kontrol ve tedavi grupları arasında istatiksel olarak anlamlı fark izlenmedi.
Sonuçlar
N-Asetilsistein ve β-glukan’ın kolitde faydalı olduğu izlendi. N-asetilsistein ve β-glukan ‘ın kombine verilmesinin ek bir fayda sağlamadığı gözlendi.

Anahtar Kelimeler

Beta-glukan, N-Asetilsistein, oksidatif stres, deneysel kolit

Destekleyen Kurum

yok

Proje Numarası

yok

Effects of N-Acetyl Cystein And Beta Glucan in Experimental Colitis Model in Rats

Öz

Introduction and Aim
Ulcerative colitis is an idiopathic and chronic disease characterized by diffuse mucosal inflammation. Oxidant and antioxidant balance is impaired in tissue. The aim of the present study was to evaluate effects of N-acetylcysteine and β-glucan against colitis.
Materials and Methods
220-250 g average, 50 male Wistar-Albino rats, were used. Single dose rectal acetic acid was applied for colitis. β-glucan 100 mg/kg/day and N-acetylycteine 200mg/kg/day were given with oral probe.
Group I (Control): Single dose rectal saline+normal diet.
Group II (Colitis): Single dose rectal acetic acid+normal diet
Group III (β-Glucan+Colitis): Single dose β-glucan 100 mg/kg (1 hour before rectal asetic acid) + Rectal acetic acid + β-glucan 100 mg / kg / day (during next 6 days).
Group IV(N-asetilsistein+Colitis): Single dose N-acetylycteine 200 mg/kg (one hour before rectal asetic acid) + Rectal acetic acid + N-acetylycteine 200 mg/kg/day (during next 6 days).
Group V (N-asetilsistein+β-glucan+Colitis): Single dose N-acetylycteine 200 mg/kg and beta-glucan 100 mg/kg/day (one hour before rectal asetic acid) + Rectal acetic acid + N-acetylycteine 200 mg/kg/day and beta-glucan 100 mg/kg/day (during next 6 days).
End of the study, distal 8 cm colon was removed. Histopathologic and biochemical assesments were carried out.
Results
MDA and MPO levels were significantly low, SOD and KAT levels were significantly higher in monotherapy and combined therapy groups. MPO and MDA levels were not statistically different between treatment groups. For other anti-oxidant enzymes (SOD, KAT), there were no difference between NAC and BG group, but in combination group anti-oxidant defence was lower than NAC group. Microscobic evaluation revealed that damage score was lowest in NAC group, but no statistical significance was found between treatment groups. Conclusions
This study suggest that N-asetilsistein and/or b-glukan treatment modalities have beneficial effect in colitis. But combined therapy compared with monotherapy did not show additional beneficial effect.

Anahtar Kelimeler

Beta-glucan, N-Acetylcysteine, oxidative stress, experimental colitis

Proje Numarası

yok

Kaynakça

• 1. Gilat T, Grossman A, Fireman Z, Rozen P. Inflammatory bowel disease in jews. ln: McConnell R, Rozen R, Langman M, Gilat T, eds. The genetics and epidemiology of inflammatory bowel disease. New York: Karger, 1986; 11: 141.
• 2. Peterson WL, Graham DY. Ulcerative colitis. In: Jewell DP, Sleisenger MH. Gastrointestinal and liver disease pathophysiology, diagnosis, management. 7th ed. Saunders; 2002; p. 2039-67.
• 3. Pietro G, Lawrance S. Epidemiology and the natural course of inflammatory bowel disease. Gastroenterol. Clin. North Am.1999; 28: 255-81.
• 4. Kirsner JB & Shorter RG. Recent developments in “nonspecific” inflammatory bowel disease. N. Engl. J. Med. 1982; 306: 775–785.
• 5. Jewell DP & Patel C. Immunology of inflammatory bowel disease. Scan. J. Gastroenterol. Suppl. 1985; 114: 119–126.
• 6. John R, Gareth A, Thomas B. Advanced Therapy of inflammatory Bowel Disease. Bayless TM. Hanauer S.B. Mucosal protective and repair agents in the treatment of colitis. 2.nd New York. 2000; 23: 107-110.
• 7. Stenson WF, Korzenik J, Yamada T, Alpers D, Kaplowitz D, Laine L. Textbook of gastroenterology. Philedelphia: Lippincott Williams Wilkins; 2003; 4 ed. p:1699-1750.
• 8. Campieri M, Gionchetti P. Bacteria as the cause of ulcerative colitis. Gut. 2001; 48: 132-5.
• 9. Asik M, Bayraktar Y. İnflamatuvar barsak hastalığında patogenez ve tedavide yenilikler. Güncel Gastroenteroloji . 1998; 2: 156–62.
• 10. Halliwell B, Gutteridge JM, Cross CE. Free radicals, antioxidants and human disease: Where are we now? J Lab. Clin. Med. 1992; 119: 598-620.
• 11. Garg R, Kumbkarni Y, Aljada A, Mohanty P, Ghanim H, Hamada W, et al. Troglitazone reduces reactive oxigen species generation by leukocytes and peroxidation and improves flow mediated vasodilatation in obese subjects. Hypertension 2000; 36: 430-5.
• 12. Cryer B, Feldman M. Cyclooxygenase-1 and cyclooxygenase-2 selectivity of widely usednonsteroidal anti-inflammatory drugs. Am. J. Med. 1998; 104: 413-21.
• 13. Çavdar C, Sifil A, Çamsarı T. Reaktif oksijen partikülleri ve antioksidan savunma. Türk Nefroloji Diyaliz ve Transplantasyon Dergisi 1997; 6: 92-5.
• 14. Esterbauer H. peroxidation products: Formation, chemical properties andbiological activities. In: Free radicals in liver injury. 1RL Press Limited, Oxford, England 1985; p: 29-47.
• 15. Yoshida N, Yoshikawa T, Naito Y, Tanigawa T, Murase H, M Kondo. A novel water soluble vitamin E derivative protects aganist experimental colitis in rat. Antioxid. Redox. Signal. 1999; 1: 555-562.
• 16. Kuralay F, Yıldız C, Özütemiz O, İslekel H, Çalıskan S, Bingol B ve ark. S. Effects of trimetazidine on acetik acid-induced colitis in female swiss rats. 2003; 2: 123-125.
• 17. Ademoglu E, Erbil YB, Tam B, Barbaros U, Ilhan E, Olgac V, ve ark. Do vitamin E and selenium have beneficial effects on trinitrobenzenesulfonic acid-induced experimental colitis. Dig. Dis. Sci. 2004; 49: 102-108.
• 18. Kim SY, Song HJ, Lee YY, et al. Biomedical Issues of Dietary fiber - Glucan. J Korean Med. Sci. 2006; 21: 781-9.
• 19. Ross GD, Vetvicka V, Yan J, et al. Therapeutic intervention with complement and beta-glucan in cancer. Immunopharmacology. 1999; 42: 61-74.
• 20. Tzianabos AO. Polysaccharide immunomodulators as therapeutic agents: structural aspects and biologic function. Clin. Microbiol. Rev. 2000; 13: 523-33.
• 21. Gu Y, Fujimiya Y, Itokawa Y, Oshima M, Choi JS, Miura T, et al. Tumoricidal effects of beta-glucans: mechanisms include both antioxidant activity plus enhanced systemic and topical immunity. Nutr Cancer. 2008; 60: 685-91.
• 22. Babayigit H, Kucuk C, Sozuer E, Yazici C, Kose K, Akgun H. Protective effect of b glucan on lung injury after cecal ligation and puncture in rats. Intensive Care Med. 2005; 865–870.
• 23. Moldeus P, Cotgreave IA, Berggren M. Lung protection by a thiolcontaining antioxidant: N-acetylcysteine. Respiration.1986; 1: 31-42.
• 24. Sener G, Toklu H, Ercan F, et al. Protective effect of beta-glucan against oxidative organ injury in a rat model of sepsis. Int. Immunopharmacol. 2000; 5: 1387–1396.
• 25. Senoglu N, Yuzbasioglu MF, Aral M, Ezberci M, Kurutas EB, Bulbuloglu E, et al. Protective Effects of N-Acetylcysteine and -Glucan Pretreatment on Oxidative Stress in Cecal Ligation and Puncture Model of Sepsis. Journal of Investigative Surgery. 2008; 21: 237- 243.
• 26. Keshavarzian A, Seghdi A, Kanofsky J et al. Excessive production of reactive oxygen metabolites by inflamed colon: Analysis by chemiluminescence probe. Gastroenterology 1992; 103: 177-185.
• 27. Cetinkaya A, Bulbuloglu E, Kurutas EB, Ciralık H, Kantarceken B, Buyukbese MA. Beneficial Effects of N-Acetylcysteine on Acetic Acid-Induced Colitis in Rats. Tohoku J. Exp. Med. 2005; 206: 131-139.
• 28. MacPherson B, MacPherson C. Experimental production of diffuse colitis in rats. Digestion 17. 1978; p: 135–150.
• 29. Yamamoto M, Yoshizaki K, Kishimoto T, and Ito H. IL–6 Is required for the development of Thl Cell–mediated murine colitis. J. Immunol. 2000; 164: 48-78.
• 30. Worthing Enzyme Manual Worthington Biochemical Corporation Freehold, New Jersey, USA, 1972.
• 31. Ohkawa D: Assay for peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979; 95: 351-358.
• 32. Beutler E. Red Cell Metabolism. A Manual of Biochemical Methods. 2nd edition. New York: Grune &Stratton Co. 1975; 261–265.
• 33. Oberley LW, Spitz DR. Assay of superoxide dismutase activity in tumor tissue. Methods Enzym measurement. 1984; 105: 457-464.
• 34. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275.
• 35. Koch TR, Yuan LX, Stryker SJ, Ratliff P, Telford GL, Opara EC: Total antioxidant capacity of colon in patients with chronic ulcerative colitis. Dig Dis Sci. 2000; 5: 1814– 1819.
• 36. Lih-Brody L, Powell SR, CollierKP, Reddy GM, Cerchia R, Kahn E, et al. Increased oxidative stress and decreased antioxidant defenses in mucosa of inflammatory bowel disease. Dig. Dis. Sci. 1996; 41: 2078 –2086.
• 37. McKenzie SJ, Baker MS, Buffinton GD, Doe WF: Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease. J. Clin. Invest. , 1996; 98: 136–141.
• 38. Grisham MB, Granger DN. Neutrophil-mediated mucosal injury. Role of reactive oxygen metabolites. Dig. Dis. Sci.1988; 33: 6-15.
• 39. Sekijuza E, Grisham MA, Li MA, Deitch EA, Granger DN. İnflammation induced intestinal hyperemia in the rat: Role of neutrophils. Gastroenterology. 1988; 95.
• 40. Fontane JC, Ward PA. Role of oxygenderived free radicals and metabolities in leukocyte dependent inflammatory reactions. Am. J. Pathol. 1982; 107.
• 41. Grisham MB. Oxidants and free radicals in inflammatory bowel disease. Lancet, 1994; 344: 859-861.
• 42. Choudhary S, Keshevarzian A, Novel antioxidants zolimid and AEOL11201 ameliorate colitis in rats. Dig. Dis. Sci. 2001; 46.
• 43. Kaya E, Özgüç ES. L-Glutamin enemas attenuate mucosal injury in experimental colitis. Dis. Colon Rectum, 1999; 42.
• 44. Pentney PT, Bubenikj GA. Melatonin reduces the severity of dextran induced colitis in mice. J.Pineal Res. 1995; 19.
• 45. Simmonds NJ, Millar AD, Blake DR. Antioxidant effects of aminosalicylates and potential new drugs for IBD: Assesment in cell-free systems and iflamed human colorectal biopsies. Aliment. Pharmacol. Therapy. 1999; 13.
• 46. Ziment I. Acetylcysteine: A drug with an interesting past and a fascinating future. Respiration. 1986; 50: 1, 26-30.
• 47. Junod AF, Jornot L, Grichting G. Comparative study on the selenium and N-acetylcystein-related effects on the toxic action of hyperoxia, paraquat and enzyme reaction hypoxanthine-xanthine oxidase in cultured endothelial cells. Agents Actions. 1987; 22: 176-183.
• 48. Fedorak RN, Empey LR, MacArthur C, Jewell LD. Misoprostol provides a colonic mucosal protective effect during acetic acid induced colitis in rats. Gastroenterology 1990; 98: 615-625.
• 49. Podolsky DK: Inflammatory bowel disease. N. Engl. J. Med. 1991; 325: 928–937.
• 50. Krawisz JE, Sharon P, Stenson WF. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Gastroenterology 1984; 87: 1344-50.
• 51. Carlson M. ; Raab, Y.; Seveus, L.; Xu, S.; Hallgren, R.; Venge, P. Human neutrophil lipocalin is a unique marker of neutrophil inflammation in ulcerative colitis and proctitis. Gut 2002, 50,501–506.
• 52. Ross GD, Vetvicka V, Yan J, Xia Y, Vetvickova J. Therapeutic intervention with complement and b-glucan in cancer. Immunopharmacology. 1999; 42: 61–74.
• 53. Vetvicka V, Yvin JC. Effect of marine b-1,3 glucan on immune reactions. Int. Immunopharmacol. 2004; 4: 721–730.
• 54. Cleary JA, Kelly GE, Husband AJ. The effectof molecular weight and b-1,6-linkages on priming of macrophage function in mice by b-(1,3)-D-glucan. Immun. Cell. Biol. 1999; 77: 395–403.
• 55. Engstad CS, Engstad RE, Olsen JO, Osteud B. The effect of soluble b-1,3-glucan and lipopolysaccharide on cytokine production and coagulation activation in whole blood. Int. Immunopharmacol. 2002; 2: 1585–1597.
• 56. Solyths J, Quinn MT. Modulation of endotoxinand enterotoxin-induced cytokine release by in vivo treatment with b-(1,6)-branched b-(1,3)-glucan. Infect. Immun. 1999; 67: 244–250.
• 57. Hashimoto T, Ohno N, Adachi Y, Yadomae T. Enhanced production of inducible nitric oxide synthase by b-glucans in mice. Immunol. Med. Microbiol. 1997; 19: 145-50.
• 58. Ljungman AG, Leanderson P, Tagesson C. Beta-(1–3)-D-glucan stimulates nitric oxide generation and cytokine mRNA Expression in macrophages. Environ. Toxicol. Pharmacol. 1998; 5: 273–281.
• 59. Babincova M, Bacova Z, Machova E, Kogan G. Antioxidant properties of carboxymethyl glucan: comparative analysis. J. Med. Food. 2002; 5: 79–83.
• 60. Krinzkova L, Durackova Z, Sandula J, Slamenova D, Sasinkova V, Sivonova M, et al. Fungal beta-(1–3)-D-glucan derivatives exhibit high antioxidative and antimutagenic activity in vitro. Anticancer. Res. 2003; 23: 2751.
• 61. Sugiyama A, Suzuki K, Mitra S, Arashida R, Yoshida E, Nakano R, et al. Hepatoprotective effects of paramylon, a beta (1-3) -D-glucan isolated from Euglena gracilis, on acute liver injury induced by carbon tetrachloride in rats. J. Vet. Med. Sci. 2009; 71: 885-890.
• 62. F Bayrak O, Turgut F, Karatas OF, Cimentepe E, Bayrak R, Catal F, et al. Oral betaglucan protects kidney against ischemia/reperfusion injury in rats. Am. J. Nephrol. 2008; 28: 190–196.
• 63. Mahgoub AA. Evaluating the prophylacticpotetial of zafirlukast against the toxic effects of acetic acid on the rat colon. Toxicol. Lett. 2003.
• 64. Kayali H, Ozdag MF, Kahraman S, Aydin A, Gonul E, Sayal A, et al. The antioxidant effect of β-Glucan on oxidative stress status in experimental spinal cord injury in rats. Neurosurg Rev. 2005; 28: 298–302.
• 65. Dong WG, Liu SP. Ameliorative effects of sodium ferulate on experimental colitis and their mechanisms in rats. World. J.Gastroentol. 2003; 9: 2533-2538.
• 66. Liu SP, Dong WG, Wu DF, Luo HS, Yu JP: Protective effect ofangelica sinensis polysaccharide on experimental immunological colon injury in rats. World J. Gastroenterol, 2003; 9: 2786–2790.
• 67. Siegers CP, Riemann D, Thies E, Younes M. Glutathion and GSH-dependent enziymes in the gastrointestinalmucosa of the rat. Cancer Lett. 1988; 40: 71-76.
• 68. Shan XQ, Aw TY, Jones DP. Glutathione-dependentprotectionagainst oxidative injury. Pharmacol. Ther. 1990; 47: 61-71.
• 69. Sener G, Eksioglu-Demiralp E, Cetiner M, Ercan F, Yegen BC. Beta-glucan ameliorates methotrexateinduced oxidative organ injury via its antioxidant and immunomodulatory effects. Eur. J. Pharmacol. 2006; 542: 170.
• 70. Sener G, Sert G, Sehirli AÖ, Arbak S, Uslu B, Gedik N, et al. Pressure ulcer-induced oxidative organ injury is ameliorated by b-glucan treatment in rats. Int. Immunopharmacol. 2006; 6: 724–732.
• 71. Sener G, Toklu H, Ercan F, Erkanli G. Protective effect of b-glucan against oxidative organ injury in a rat model of sepsis, Int. Immunopharmacol. 2005; 5: 1387–1396.