In Vitro Genotoxic Effects of Ursodeoxycholic Acid in Human Peripheral Blood Lymphocytes

Öz

Ursodeoxycholic acid (UDCA), the 7b epimer of chenodeoxycholic acid has been increasingly

used for the treatment of cholestatic liver diseases. In this study, it was aimed to determine the potential in vitro

genotoxic effects of ursodeoxycholic acid in human peripheral blood lymphocytes. The potential genotoxic and

cytotoxic effects of UDCA were investigated in vitro by using chromosome aberration and mitotic index assays.

Human peripheral blood lymphocytes were treated with 10, 50 and 100 μg/ml of UDCA for 24 and 48 hours. The

data were evaluated using one-way ANOVA (Post Hoc Analysis-LSD Test) test in the SPSS statistics program.

Statistical results showed that the applied doses of UDCA did not cause a significant increase in chromosome

aberration frequency, and also it did not cause a significant decrease in mitotic index when compared to the control

(p>0.05). The finding that UDCA does not cause an increase in chromosomal aberrations may be related to the

production of this substance physiologically as bile acid in the human body, even in trace amount. The fact that

there are many patients using UDCA increases the importance of findings obtained from this study.

Anahtar Kelimeler

• Chromosome aberration test,Genotoxic effect,Human peripheral lymphocytes,Mitotic index,Ursodeoxycholic acid

Ursodeoksikolik Asit’in İnsan Periferal Kan Lenfositlerindeki in Vitro Genotoksik Etkisi

Öz

Kenodeoksikolik asitin 7b epimeri olan ursodeoksikolik asit (UDKA), kolestatik karaciğer hastalıklarının

tedavilerinde artan bir şekilde kullanılmaktadır. Bu çalışmada, Ursodeoksikolik asitin, insan periferal kan

lenfositlerindeki in vitro sitotoksik ve genotoksik etkilerinin belirlenmesi hedeflendi. UDKA’nın potansiyel

genotoksik ve sitotoksik etkisi, kromozom aberasyon ve mitotik indeks testleri kullanılarak in vitro olarak araştırıldı.

İnsan peripheral kan lenfositleri, 24 ve 48 saat süreyle, 10, 50 ve 100 μg/ml UDKA ile muamele edildi. Veriler

SPSS istatistik programında, Tek Yönlü Anova (Post Hoc Analiz-LSD Test) testi ile analiz edildi. Elde edilen

istatistik sonuçları, kontrolle karşılaştırıldığında, uygulanan UDKA konsantrasyonlarının mitotik indeks değerlerini

düşürmediğini ve kromozom anomali frekanslarında da önemli bir artışa neden olmadığını göstermektedir (p>0.05).

UDKA’nın insan kromozomlarında anomalileri artırmaması bulgusu, bu maddenin az da olsa insan vücudunda

fizyolojik olarak üretilen bir safra asiti olması ile bağlantılı olabilir. UDKA’yı kullanan pek çok hasta olması, bu

çalışmadan elde ettiğimiz bulguların önemini artırmaktadır.

Anahtar Kelimeler

• Genotoksik etki,İnsan periferal lenfositleri,Kromozom aberasyon testi,Mitotik indeks,Ursodeoksikolik asit

Kaynakça

1. Beuers U, Boyer JL, Paumgartner G, 1998. Ursodeoxycholic Acid in Cholestasis: Potential Mechanisms of Action and Therapeutic Applications. Hepatology, 28(6): 1449-1453.
2. Burnat G, Majka J, Konturek PC, 2010. Bile acids are multifunctional modulators of the Barrett’s carcinogenesis. J. Physiol. Pharmacol., 61: 185–192.
3. Dodo M, Owen RW, Thompson MH, Hill MJ, 1984. A comparison of the effects of chenodeoxycholic acid and ursodeoxycholic acid treatment on faecal bile acid profiles in healthy subjects. Biochem. SOC. Trans., 12: 862-863.
4. El-Sherbiny GA, Taye A, Abdel-Raheem IT, 2009. Role of ursodeoxycholic acid in prevention of hepatotoxicity caused by amoxicillin-clavulanic acid in rats. Ann Hepatol., 8(2):134-40.
5. Evans HJ, Kilbey BJ, Legator M, Nicohols W, Ramel C (1984) Handbook of Mutagenicity Test Procedures. Human Peripheral Blood Lymphocytes for The Analysis of Chromosome Aberrations in Mutagen Tests. Elsevier Science Publishers, BV, 405-406.
6. Fimognari C, Nüsse M, Cesari R, Forti GC, and Hrelia P, 2001. Micronuklei induction, cell cycle delay and apoptosis as markers of cellular stress caused by ursodeoxycholic acid in human lymphocytes. Mutation Research, 495: 1-9.
7. Galle PR, Theilmann L, Raedsch R, Otto G, Stiehl A, 1990. Ursodeoxycholate reduces hepatotoxicity of bile salts in primary human hepatocytes. Hepatology, 12 (3 Pt 1):486-91.
8. Glantz L, Avramovich A, Trembovler V, Gurvitz V, Kohen R, Eidelman LA, Shohami E, 2005. Ischemic preconditioning increases antioxidants in the brain and peripheral organs after cerebral ischemia. Exp. Neurol., 192: 117–124.

9. Guarino MPL, Altomore A, Cocca S, Emerenziani S, Cicala M, 2013. Ursodeoxycholic acid therapy in gallbladder disease, a story not yet completed. World J Gastroenterol, 19(31): 5029-5034.
10. Heuman DM, Bajaj R, 1994. Ursodeoxycholate conjugates protect against disruption of cholesterol-rich membranes by bile salts. Gastroenterology, 106: 1333-41.
11. Higashi H, Setoguchi T, and Katsuki T, 1978. Interconversion between chenodeoxycholic acid and ursodeoxycholic acid in anaerobic cultures of intestinal bacteria and reduction of 7-ketolithocholic acid to both bile acids. Acta Hepatol. Jpn. 19: 803.
12. Hofmann AF, 2011. Herbert Falk: a vital force in the renaissance of bile acid research and bile acid therapy. Dig Dis. Sci.,29: 23-3.
13. Hofmann AF, Hagey LR, 2008. Bile acids: chemistry, pathochemistry, biology, pathobiology, and therapeutics. Cell. Mol. Life Sci. 65: 2461–2483.
14. Im EO, Choi YH, Paik KJ, Suh H, Jin Y, Kim KW, Yoo YH, Kim ND, 2001. Novel bile acid derivatives induce apoptosis via a p53-independent pathway in human breast carcinoma cells. Cancer Lett, 163: 83-93.
15. Ishizaki K, Imada T, Tsurufuji M, 2005. Hepatoprotective bile acid 'ursodeoxycholic acid (UDCA)' Property and difference as bile acids.Hepatol Res., 33(2):174-7.
16. Kotb MA, 2009. Ursodeoxycholic acid in neonatal hepatitis and infantile paucity of intrahepatic bile ducts: Review of a historical cohort. Dig. Dis. Sci., 54: 2231–2241.
17. Kotb MA, 2012. Molecular Mechanism of Ursodeoxycholic Acid Toxity & Side Effects: Ursodeoxycholic Acid Freezes Regeneration & Induces Hibernation Mode. Molecular Sciences, 13: 8882-8914.
18. Lim AG, Jazrawi RP, Northfield TC, 1995. The ursodeoxycholic acid story in primary biliary cirrhosis. Gut, 37 (3): 301-304.
19. Lindor KD, Kowdley KV, Luketic VAC, Harrison ME, McCashland T, Befeler AS, Harnois D, Jorgensen R, Petz J, Keach J et al., 2009. High dose ursodeoxycholic acid for the treatment of primary sclerosing Cholangitis. Hepatology, 50: 808–814.
20. Liu H, Oin CY, Han GO, Xu HW, Meng M, Yang Z, 2007. Mechanism of apoptotic effects induced selectively by ursodeoxycholic acid on human hepatoma cell lines. World J Gastroenterol, 13 (11): 1652-1658.
21. Lukivskaya O, Zavodnik L, Knas M, Buko V, 2006. Antioxidant mechanism of hepatoprotection by ursodeoxycholic acid in experimental alcoholic steatohepatitis. Adv Med Sci., 51:54-9.
22. Makino I, Tanaka H, 1998. From a choleretic to an immunomodulator: Historical review of ursodeoxycholic acid as a medicament. Journal of Gastroenterology and Hepatology, 13: 659-664.
23. Owen RW, Wait R, Bilton RF, 1988. Biotransformation of ursodeoxycholic acid by Pseudomonas sp NCIB 10590. Journal of Lipid Research, 29: 459-468.
24. Pusl T, Beuers U, 2006. Ursodeoxycholic acid treatment of vanishing bile duct syndromes. World J Gastroenterol, 12(22): 3487-3495.
25. Roma MG, Toledo FD, Boaglio AC, Basiglio CL, Crocenzi FA, 2011. Sánchez Pozzi EJ.Ursodeoxycholic acid In cholestasis: linking action mechanisms to therapeutic applications. Clin Sci (Lond).,121(12):523-44.
26. Salvioli G, Salati R, Lugli R, Zanni C, 1983. Medical treatment of biliary duct stones: effect of ursodeoxycholic acid administration. Gut, 24: 609-614.
27. Song P, Zhang Y, and Klaassen CD, 2011. Dose-Response of Five Bile Acids on Serum and Liver Bile Acid Concentrations and Hepatotoxicty in Mice. Toxicological Sciences, 123(2): 359–367.