- P-ISSN 1010-0695
- E-ISSN 2288-3339
Objectives: Ssanghwa-tang (SHT) is a traditional herbal formula comprising nine medicinal herbs, and it is used for reducing fatigue in Korea. SHT exerts various effects such as anti-inflammatory, antioxidant, and anti-aging activities, and protection against acute hepatotoxicity. However, the genotoxicity of SHT has not yet been established. Methods: Ten components were identified in SHT water extract by using high-performance liquid chromatography analysis. We assessed the genotoxicity of SHT by using bacterial reverse mutation (Ames test), chromosome aberration, and in vivo micronucleus tests. Results: The contents of paeoniflorin, glycyrrhizin, and liquiritin apioside in SHT were 15.57, 6.94, and 3.48 mg/g extract, respectively. SHT did not increase the revertant colonies of Salmonella typhimurium and Escherichia coli strains in the presence or absence of metabolic activity. Although SHT did not induce structurally abnormal chromosomes in Chinese hamster lung (CHL) cells in the presence of metabolic activity, the number of structurally aberrated chromosomes increased dose-dependently in the absence of metabolic activity. In the in vivo micronucleus test, SHT did not affect the formation of micronuclei compared with the vehicle control. Conclusions: Genotoxicity of SHT was not observed in the Ames test and in vivo micronucleus test. However, based on the results of chromosome aberration test, it can be presumed that SHT has the potential to induce genotoxicity because it induced structurally abnormal chromosomes in the absence of metabolic activity.
1. World Health Organization (WHO). WHO guidelines on safety monitoring of herbal medicines in pharmacovigilance systems. 2004. Available from: http://www.who.int/iris/handle/10665/43034/
2. Firenzuoli F, Gori L. Herbal medicine today:clinical and research issues. Evid Based Complement Alternat Med. 2007;4(S1):37-40.
3. Wang J, van der Heijden R, Spruit S, Hankermeier T, Chan K, van der Greef J, et al. Quality and safety of Chinese herbal medicines guided by a systems biology perspective. J Ethnopharmacol. 2009;126(1):31-41.
4. Asif M. A brief study of toxic effects of some medicinal herbs on kidney. Adv Biomed Res. 2012;1: 44.
5. Ekor M. The growing use of herbal medicines:issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol. 2014;4:177.
6. Stournaras E, Tziomalos K. Herbal medicine -related hepatotoxicity. World J Hepatol. 2015;7(19):2189-93.
7. Cho JH, Oh DS, Hong SH, Ko H, Lee NH, Park SE, et al. A nationwide study of the incidence rate of herb-induced liver injury in Korea. Arch Toxicol. 2017;91(12):4009-15.
8. Korea Pharmaceutical and Bio-Pharma Manufacturers Association (KPBMA). Production achievements for pharmaceuticals, etc. in 2018. Seoul: KPBMA. 2019.
9. Shim KS, Lee JH, Ma CJ, Lee YH, Choi SU, Lee J, et al. Inhibitory effect of Ssanghwa-tang on bone loss in ovariectomized rats. Anim Cells Syst. 2010;14(4):283-9.
10. Lee JH, Yang MC, Shim KS, Lee JH, Ma JY. Protective effect of Ssanghwa-tang fermented by Lactobacillus fermentum against carbon tetrachloride-induced acute hepatotoxicity in rats. Afr J Tradit Complement Altern Med. 2011;8(3):312-21.
11. Kim A, Yim NH, Im M, Jung YP, Liang C, Cho WK, et al. Ssanghwa-tang, an oriental herbal cocktail, exerts anti-melanogenic activity by suppression of the p38 MAPK and PKA signaling pathways in B16F10 cells. BMC Complement Altern Med. 2013;13:214.
12. Kim IH, Hwang GJ. Studies on the anti -inflammatory activities of 'Ssangwha-Tang'. Korean J Pharmacogn. 1981;12(3):131-5.
13. Han DS, Lee HK, Cho HJ. Analgesic and anticonvulsionary effects of Ssanghwa-Tang. Korean J Pharmacogn. 1983;14(2):60-3.
14. Park JY, Hwang JG, Yun JK, Han KH, Do EJ, Kim SO, et al. Effect of ssanghwa-tang extract on antioxidant and anti-aging enzyme activities. Korea J Herbol. 2012:27(3), 67-74.
15. Ko JW, Shin NR, Park SH, Lee IC, Ryu JM, Cho YK, et al. Ssanghwa-Tang, a traditional herbal formula, suppresses cigarette smoke -induced airway inflammation via inhibition of MMP-9 and Erk signaling. Mol Cell Toxicol. 2017;13(3):295-304.
16. Kim SJ, Lee MY, Shin IS, Seo CS, Ha H, Huh JI, et al. Single dose acute toxicity of Ssanghwa-tang in Crl:CD (SD) rats. Korea J Herbol. 2011;26(2):39-43.
17. Yoo SR, Ha H, Lee MY, Shin HK, Han SC, Seo CS. A 4-week repeated oral dose toxicity study of Ssanghwa-tang in Crl:CD Sprague Dawley rats. Evid Based Complement Alternat Med. 2019;2019: 2135351.
18. Organization for Economic Cooperation and Develppment (OECD) Guidelines for the Testing of Chemicals. ‘Bacterial reverse mutation test’ (OECD TG#471). 1997. Available from: https://www.oecd.org/chemicalsafety/risk -assessment/1948418.pdf
19. OECD Guidelines for the Testing of Chemicals.. ‘In vitro mammalian chromosome aberration test’ (OECD TG#473). 2014. Available from:https://ntp.niehs.nih.gov/iccvam/suppdocs/fed docs/oecd/oecd-tg473-2014-508.pdf
20. OECD Guidelines for the Testing of Chemicals. ‘Mammalian erythrocyte micronucleus test’(OECD TG#474). 2016. Available from:https://www.oecd-ilibrary.org/docserver/9789264264762-en.pdf?expires=1611031436&id=id &accname=guest&checksum=95E219EF93374249AE0BAB4C1ED1EA04
21. Maron DM, Ames BN. Revised methods for the Salmonella mutagenicity test. Mutat Res. 1983;113(3-4):173-215.
22. Ishidate M. Chromosomal aberration tests in vitro as a primary screening tool for environmental mutagens and /or carcinogens. GANN Monogr Canver Res. 1981;27:95-108.
23. Dean BJ. Assays for the detection of chemically-induced chromosome damage in cultured mammalian cells. In: Venitt S, Parry JM, editors. Mutagenicity testing, a practical approach. Washington DC:IRL Press. 1984.
24. Galloway SM, Aardema MJ, Ishidate M Jr, Ivett JL, Kirkland DJ, Morita T, et al. Report from working group on in vitro tests for chromosomal aberrations. Mutat Res. 1994;312(3):241-61.
25. Japanese Environmental Mutagen Society -Mammalian Mutagenicity Study Group. Atlas of chromosome aberration by chemicals. Tokyo:JEMS-MMS. 1988.
26. Gollapudi B, Kamra OP. Application of a simple giemsa-staining method in the micronucleus test. Mutat Res. 1979;64(1):45-6.
27. Van Miert A.S.J.P.A.M. The use in animals of drugs licensed for human use only. In:Van Miert A.S.J.P.A.M., Bogaert M.G., Debackere M. (Eds.), 1986. Comparative Veterinary Pharmacology, Toxicology and Therapy. MTP Press, Boston, pp.489–500.
28. Mortelmans K, Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutat Res. 2000;455(1-2):29-60.
29. Clare G. The in vitro mammalian chromosome aberration test. In: Parry JM, Parry EM, editors. Genetic toxicology. New York:Springer. 2012.
30. Luzhna L, Kathiria P, Kovalchuk O. Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond. Front Genet. 2013;4:131.
31. Kim DH, Kim NJ, Jang JB, Song BY. Studies on the effects of herbal medicines on the fetus during pregnancy (II)-Mutagenesis and chromosomal aberration of herbal medicines-. J Korean Orient Med. 1999;20(2):121-7.
32. Hilliard CA, Armstrong MJ, Bradt CI, Hill RB, Greenwood SK, Galloway SM. Chromosome aberrations in vitro related to cytotoxicity of nonmutagenic chemicals and metabolic poisons. Environ Mol Mutagen. 1998;31: 316-26.
33. Yamamura E, Aruga C, Muto S, Baba N, Uno Y. Correlation between the results of in vitro and in vivo chromosomal damage tests in consideration of exposure levels of test chemicals. Genes Environ. 2018;40:6.