The cyto- and genotoxic potantials of the herbal mixtures frequently used in Turkey

Abstract

Despite the profound therapeutic advantages possessed by herbs, some constituents of herbs have been shown to be potentially toxic. Knowledge on their safety is inadequate although the concerns have been raised over the lack of quality controls. We aimed to determine safety of six herbal mixtures used as tea, included forty plant species, chosen on the basis of their frequency of medicinal use and commercial importance in Turkey. Their cytotoxic activities were evaluated by determining mitochondrial succinate dehydrogenase (XTT) and extracellular lactate dehydrogenase (LDH) activities on human cerviks cell line (HeLa). For their genotoxic activities, two bacterial mutation assays, Ames assay with Salmonella typhimurium TA98 and TA100 strains and umu assay with S. thyphimurium TA1535/pSK1002 strain, were used. The 50% inhibition concentration (IC) values of the extracts for LDH and XTT tests were 6.52-63.53 and 18.75-104.67 mg/mL, respectively. In the genotoxicity studies conducted by umu assay, no extracts possessed genotoxic activities at 6.25-25 mg/mL. On the contrary, Ames bacterial mutagenicity assay conducted at the same concentrations revealed that (i) some extracts were shown mutagenic activities with metabolic activation (ii) TA100 strain was more sensitive than TA98 strain to the extracts, (iii) especially, three herbal mixtures may include ingredients shown mutagenic activities both in two strains and with metabolic activation. Our findings showed that herbal teas have some significant toxic effects. Therefore, the researchers and/or national authorities should consider that the use of herbal products may be harmful to human health

Keywords

• herbal mixtures
• Ames assay
• umu assay
• LDH test
• XTT test

References

1. WHO. (1996). Technical Report Series 863:178-184. Available at www.apps.who. int/medicinedocs/en/d/Js5516e/.
2. Maiti B, Nagori BP, Singh R, et al. (2011). Recent trends in herbal drugs: A review. Int J Drug Res Tech 1(1):17-25.
3. Akinboro A, Bakare AA. (2007). Cytotoxic and genotoxic effects of aqueous extracts of five medicinal plants on Allium cepa Linn. J Ethnopharmacol 112:470-475.
4. Ernst E. (2004). Risks of herbal medicinal products. Pharmacoepidemiol Drug Safety 13:767-771.
5. Rietjens IMCM, Boersma MG, Van der Woude H, et al. (2005). Flavonoids and alkenylbenzenes: mechanisms of mutagenic action and carcinogenic risk. Mutat Res 574:124-138.
6. Haugen DA, Peak MJ. (1983). Mixtures of polycyclic aromatic compounds inhibit mutagenesis in the Salmonella microsome assay by inhibition of metabolic activation. Mutat Res 116:257-269.
7. Hermann M. (1981). Synergistic effects of individual polycyclic aromatic hydrocarbons on the mutagenicity of their mixtures. Mutat Res 90:399-409.
8. Firenzuoli F, Gori L. (2007). Herbal medicine today: clinical and research issues. J Evid Based Complement Altern Med 4:37-40.

9. Rousseaux CG, Schachter H. (2003). Regulatory issues concerning the safety, efficacy and quality of herbal remedies. Birth Defects Res B Dev Reprod Toxicol 68:505-510.
10. Bandaranayake WM. (2006). Quality control, screening, toxicity, and regulation of herbal drugs.Modern Phytomedicine. Turning Medicinal Plants into Drugs. Edited by I. Ahmad, F. Aqil, and M. Owais, 25-57.
11. Bast A, Chandler RF, Choy PC, et al. (2002). Botanical health products, positioning and requirements for effective and safe use. Environ Toxicol Pharmacol 12:195-211.
12. EMEA Ad hoc Working Group on Herbal Medicinal Products. (1999). Report from the ad hoc Working Group on Herbal Medicinal Products (EMEA/HMPWG/25/99). Working Group on Herbal Medicinal Products, London.
13. Jordan SA, Cunningham DG, Marles RJ. (2010). Assessment of herbal medicinal products: Challenges, and opportunities to increase the knowledge base for safety assessment. Toxicol Appl Pharmacol 243:198-216.
14. Cakilcioglu U, Turkoglu I. (2010). An ethnobotanical survey of medicinal plants in Sivrice (Elazığ-Turkey). J Ethnopharmacol 132:165-175.
15. Kültür Ş. (2007). Medicinal plants used in Kırklareli Province (Turkey). J Ethnopharmacol 111:341-364.
16. Roehm NW, Rodgers GH, Hatfield SM, et al. (1991). An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immun Methods 142:257-265.
17. Legrand C, Bour JM, Jacob C, et al. (1992). Lactate dehidrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker. J Biotechnol 25:231-243.
18. Hakamura A, Shimada H, Nakajima M, et al. (2005). Salmonella/human S9 mutagenicity test: A collaborative study with 58 compounds. Mutagenesis 20(3):217- 228.
19. Umbuzeiro GA, Rech CM, Correia S, et al. (2010). Comparison of the Salmonella/ microsome microsuspension assay with the new microplate fluctuation (MPF) protocol for testing the mutagenicity of environmental samples. Environ Mol Mutagen 51:31-38.
20. Flückiger-Isler S, Kamber M. (2006). The Ames MPF™ 98/100 Assay: Novel mutagenicity testing in liquid microplate format using S. typhimurium TA98 and TA100. EEMS Prague.
21. Oda Y, Nakamuro S, Oki I, et al. (1985). Evaluation of the new system (umu-test) for the detection of environmental mutagens and carcinogens. Mutat Res 147:219-229.
22. Alves AM, Vidal LS, Kuster RM, et al. (2009). Genotoxic and mutagenic effects of Melissa officinalis (Erva Cidreira) extracts. The Open Toxicol J 3:58-69.
23. Maron DM, Ames BN. (1983). Revised methods for the Salmonella mutagenicity test. Mutat Res 113:173-215.
24. Reifferscheid G, Heil J, Oda Y, et al. (1991). A microplate version of the SOS/umu- test for rapid detection of genotoxic potentials of environmental samples. Mutat Res 253:215-222.
25. Zeiger E. (2001). Mutagens that are not carcinogenic: faulty theory or faulty tests? Mutat Res 492:29-38.
26. Ahmad N, Mukhtar H. (1999). Green tea polyphenols and cancer: Biologic mechanisms and practical implications. Nutrition Rev 57(3):78-83.
27. Katiyar SK, Mukhtar H. (1996). Tea in chemoprevention of cancer: epidemiologic and experimental studies. Int J Oncol 8:221-238.
28. Kruawan K, Kangsadalampai K. (2006). Antioxidant activity, phenolic compound contents and antimutagenic activity of some water extract of herbs. Thai J Pharm Sci 30:28-35.
29. Mimica-Dukić N, Bugarin D, Grbović S, et al. (2010). Essential oil of Myrtus communis L. as a potential antioxidant and antimutagenic agents. Mol 15:2759-2770.
30. Rosa MR, Melecchi MIS, Halmenschlager RC, et al. (2006). Antioxidant and Antimutagenic Properties of Hibiscus Tiliaceus L. Methanolic Extract. J Agric Food Chem 54(19):7324-7330.
31. de Carvalho NC, Correa-Angeloni MJF, Leffa DD, et al. (2011). Evaluation of the genotoxic and antigenotoxic potential of Melissa officinalis in mice. Gen Mol Biol 34(2):290-297.
32. Higashimoto M, Purintrapiban J, Kataoka K, et al. (1993). Mutagenicity and antimutagenicity of extracts of three spices and a medicinal plant in Thailand. Mutat Res 303:135-142.
33. Mahmoud I, Alkofahi A, Abdelaziz A. (1992). Mutagenic and toxic activities of several spices and some Jordanian medicinal plants. Int J Pharmacog 30:81-85.
34. Ebeed NM, Abdou HS, Booles HF, et al. (2010). Antimutagenic and chemoprevention potentialities of sweet fennel (Foeniculum vulgare Mill.) hot water crude extract. J Am Sci 6(9):831-842.
35. Saadat M, Masoudi M, Zendehboody Z. (2007). Genotoxicity of Gasterolan (An Herbal Product) on chromosomes of cultured human lymphocytes and rat bone marrow. J Pharmacol Toxicol 2(3):304-306.
36. Mitscher LA, Drake S, Gollapudi SR, et al. (1986). Isolation and identification of higher plant agents active in antimutagenic assay systems: Glycyrrhiza glabra. Basic Life Sci 39:153-165.
37. Zani F, Cuzzoni MT, Daglia M, et al. (1993). Inhibition of mutagenicity in Salmonella typhimurium by Glycyrrhiza glabra extract, glycyrrhizinic acid, 18a- and 18b-glycyrrhetinic acids. Planta Medica 59:502-507.
38. Martinez A, Ikken Y, Cambero MI, et al. (1999). Mutagenicity and cytotoxicity of fruits and vegetables evaluated by the Ames test and 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay. Food Sci Technol Int 5:431-437.
39. Chang PY, Wang CK, Liang CT, et al. (1964). The pharmacological action of Zang Hong Hua (Crocus sativus L.). Effects on the uterus and/or strous cycle. Yao Hsueh Hsueh Pao 11:94-100.
40. Escribano J, Diaz-Guerra MJ, Riese HH, et al. (2000). The cytotoxic effect of glucoconjugate extracted from corms of saffron plant (Crocus sativus) on human cell lines in culture. Planta Medica 66:157-162.
41. Nair SC, Kurumboor SK, Hasegawa JH. (1995). Saffron chemoprevention in biology and medicine: A review. Cancer Biother Radiopharm 10:257-264.
42. Abdullaev FI, Riveron-Negrete L, Caballero-Ortega H, et al. (2003). Use of in vitro assays to assess the potential antigenotoxic and cytotoxic effects of saffron (Crocus sativus L.). Toxicol in Vitro 17:1-6.
43. Ajami M, Eghtesadi S, Pazoki-Toroudi H, et al. (2010). Effect of Crocus sativus on gentamicin induced nephrotoxicity. Biol Res 43:83-90.
44. Sousa SM, Silva PS, Viccini LF. (2010). Cytogenotoxicity of Cymbopogon citratus (DC) Stapf (lemon grass) aqueous extracts in vegetal test systems. An Acad Bras Cienc 82(2):305-311.
45. Yang G, Zhong L, Jiang L, et al. (2010). Genotoxic effect of 6-gingerol on human hepatoma G2 cells. Chemico-Biol Interact 185:12-17.