In Vivo Investigation of Genotoxic and Antigenotoxic Potential of Hypericum Perforatum Extract in Rats

Öz

This study was planned to reveal in vivo genotoxic, antigenotoxic and cytotoxic properties of methanolic extracts of St. John's Wort plant (Hypericum perforatum). The methanol extract of H. perforatum was administrated to rats alone or in combination with N-nitrosodimethylamine and Triclosan (positive control). The concentrations of H. Perforatum were 20, 100, 500 mg/kg and the period of administration was 30 days. At the end of this period, chromosome aberration test used to determine genotoxic and antigenotoxic effects of H. perforatum. In addition, total oxidant and antioxidant levels of peripheral blood serum of rats were determined by spectrophotometric method. According to our result, H. perforatum extract did not demonstrate genotoxic effect at concentrations administrated. When compared to the negative control, the percentage of abnormal cells was increased in groups treated with test substance, N-nitrosodiethylamine and triclosan together. However, these increases were not as higher as groups treated with N-nitrosodiethylamine plus triclosan (positive control). Even, in groups treated with the test substance additional N-nitrosodiethylamine plus triclosan, the ratio of abnormal cells was inversely proportional to the increase in the concentration. The lowest ratio of abnormal cells was in the group in which the highest concentration of the H. perforatum (500 mg/kg) was applied. The highest two concentrations of the H. perforatum (100 and 500 mg/kg) significantly increased the mitotic index. In the group treated with test substance and positive control together, the mitotic index was even greater. At the same time 100 mg/kg of H. perforatum extract significantly reduced the oxidative stress.

Anahtar Kelimeler

• Hypericum perforatum,N-nitrosodiethylamine,triclosan,cytotoxicity,genotoxicity,rat bone marrow,oxidative stress

Kaynakça

1. Öztürk Y. Testing the antidepressant effects of Hypericum species on animal models. Pharmacopsychiatry. 1997; 30(S 2): 125-128.
2. Baytop T, Türkiye’de Bitkiler ile Tedavi Geçmiste ve Bugün [Treatment Plant in Turkey; Past and Present]. Nobel Tıp Kitabevi, İlaveli İkinci Baskı, İstanbul. Turkish, 1999.
3. Ernst E, Hypericum: the genus Hypericum. CRC Press. 2003.
4. Klemow KM, Bartlow A, Crawford J, Kocher N, Shah J, Ritsick M. Medical Attributes of St. John's Wort (Hypericum perforatum). Herbal Medicine: Biomolecular and Clinical Aspects. I. F. F. Benzie and S. Wachtel-Galor. Boca Raton (FL), 2011.
5. Wolfle U, Seelinger G, Schempp CM. Topical application of St. John's wort (Hypericum perforatum). Planta Med. 2014; 80(2-3): 109-120.
6. Okpanyi SN, Lidzba H, Scholl BC, Miltenburger HG. Genotoxicity of a standardized Hypericum extract. Arzneimittelforschung. 1990; 40(8): 851-855.
7. Ramos AA, Marques F, Fernandes-Ferreira M, Pereira-Wilson C. Water extracts of tree Hypericum sps. protect DNA from oxidative and alkylating damage and enhance DNA repair in colon cells. Food Chem Toxicol. 2013; 51: 80-86.
8. Hostanska K, Reichling J, Bommer S, Weber M, Saller R. Hyperforin a constituent of St John's wort (Hypericum perforatum L.) extract induces apoptosis by triggering activation of caspases and with hypericin synergistically exerts cytotoxicity towards human malignant cell lines. Eur J Pharm Biopharm. 2003; 56(1): 121–32.

9. Roscetti G, Franzese O, Comandini A, Bonmassar E. Cytotoxic activity of Hypericum perforatum L. on K562 erythroleukemic cells: differential effects between methanolic extract and hypericin. Phytother Res. 2004; 18(1): 66-72.
10. Schmitt LA, Liu Y, Murphy PA, Petrich JW, Dixon PM, Birt DF. Reduction in hypericin-induced phototoxicity by Hypericum perforatum extracts and pure compounds. J Photochem Photobiol B. 2006; 85(2): 118-130.
11. Tepkeeva II, Aushev VN, Zborovskaya IB, Demushkin VP. Cytostatic activity of peptide extracts of medicinal plants on transformed A549, H1299, and HeLa Cells. Bull Exp Biol Med. 2009; 147(1): 48-51.
12. Bartsch H, Montesano R. Relevance of nitrosamines to human cancer. Carcinogenesis. 1984; 5(11): 1381-1393.
13. Bogovski P, Bogovski S. Special report animal species in which n‐nitroso compounds induce cancer. Int J Cancer. 1981; 27(4): 471-474.
14. Mirvish SS. Role of N-nitroso compounds (NOC) and N-nitrosation in etiology of gastric, esophageal, nasopharyngeal and bladder cancer and contribution to cancer of known exposures to NOC. Cancer letters. 1995; 93(1): 17-48.
15. Levy CW, Roujeinikova A, Sedelnikova S, Baker PJ, Stuitje AR, Slabas AR, Rice DW, Rafferty JB. Molecular basis of triclosan activity. Nature. 1999; 398(6726): 383.
16. Yueh MF, Taniguchi K, Chen S, Evans RM, Hammock BD, Karin M, Tukey RH. The commonly used antimicrobial additive triclosan is a liver tumor promoter. Proc Natl Acad Sci U S A. 2014; 111(48): 17200-5
17. Schempp CM, Kirkin V, Simon-Haarhaus B, Kersten A, Kiss J, Termeer CC, Simon JC. Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John's wort that acts by induction of apoptosis. Oncogene. 2002; 21(8): 1242.
18. Silva SM, Martinho A, Moreno I, Silvestre S, Granadeiro LB, Alves G, Duarte AP, Domingues F, Gallardo E. Effects of Hypericum perforatum extract and its main bioactive compounds on the cytotoxicity and expression of CYP1A2 and CYP2D6 in hepatic cells. Life Sci. 2016; 144: 30-6.
19. Rietjens IM, Martena MJ, Boersma MG, Spiegelenberg W, Alink GM. Molecular mechanisms of toxicity of important food-borne phytotoxins. Mol Nutr Food Res. 2005; 49(2): 131-158.
20. Miadokova E, Chalupa I, Vlckova V, Sevcovicova A, Nadova S, Kopaskova M, Hercegova A, Gasperova P, Alfoldiova L, Komjatiova M, Csanyiova Z. Galova E, Cellarova E, Vlcek D. Genotoxicity and antigenotoxicity evaluation of non-photoactivated hypericin. Phytother Res. 2010; 24(1): 90-5.
21. Peron AP, Mariucci RG, de Almeida IV, Düsman E, Mantovani MS, Vicentini VE. Evaluation of the cytotoxicity, mutagenicity and antimutagenicity of a natural antidepressant, Hypericum perforatum L. (St. John's wort), on vegetal and animal test systems. BMC Complement Altern Med. 2013; 6(13): 97. Lu YH, Du CB, Liu JW, Hong W, Wei DZ. Neuroprotective effects of Hypericum perforatum on trauma induced by hydrogen peroxide in PC12 cells. Am J Chin Med. 2004; 32(3): 397-405.
22. Zou YP, Lu YH, Wei DZ. Protective effects of a flavonoid-rich extract of Hypericum perforatum L. against hydrogen peroxide-induced apoptosis in PC12 cells. Phytother Res. 2010; 24 Suppl 1: S6-S10.
23. Cinci L, Di Cesare Mannelli L, Maidecchi A, Mattoli L, Ghelardini C. Effects of Hypericum perforatum extract on oxaliplatin-induced neurotoxicity: in vitro evaluations. Zeitschrift fur Naturforschung. C, Journal of Biosciences. 2017; 72, 219-226.
24. Polyplant. https://www.centerchem.com/Products/DownloadFile.aspx?FileID=7270) accessed on 28 June 2017 OECD Test No. 475: Mammalian Bone Marrow Chromosomal Aberration Test, OECD Publishing.
25. Timocin T, Ila HB. Investigation of flurbiprofen genotoxicity and cytotoxicity in rat bone marrow cells. Drug Chem Toxicol. 2015; 38(3): 355-360. Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem. 2005; 38(12): 1103-1111.
26. Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem. 2004; 37(4): 277-285.
27. Harma M, Harma M, Erel O. Increased oxidative stress in patients with hydatidiform mole. Swiss Med Wkly. 2003; 133(41-42): 563-566. Kosecik M, Erel O, Sevinc E, Selek S. Increased oxidative stress in children exposed to passive smoking. Int J Cardiol. 2005; 100(1): 61-64.
28. Yumru M, Savas HA, Kalenderoglu A, Bulut M, Celik H, Erel O. Oxidative imbalance in bipolar disorder subtypes: a comparative study. Prog Neuropsychopharmacol Biol Psychiatry. 2009; 33(6): 1070-1074.