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Evaluation of in vitro and in vivo genotoxicity of Angelica acutiloba in a standard battery of assays

Abstract

Among three representative species of Angelica found in Asian countries, including Korea, China, and Japan, Angelica acutiloba (AA) has been used as traditional herbal medicine with antitumor, antiinflammatory, anti-obesity, and anti-diabetes activities. In this study, the potential genotoxicity and mutagenicity of the AA extract were examined in a battery of in vitro and in vivo tests (bacterial reverse mutation assay, in vitro chromosomal aberrations assay, and in vivo micronucleus assay) in accordance with the test guidelines for toxicity testing developed by the Organization for Economic Cooperation and Development. Upon testing in the bacterial mutation assay (Ames test) using five Salmonella typhimurium TA98, TA100, TA102, TA1535 and TA1537, no significant increase the number of revertant colonies in the metabolic activation system and non-activation system was noted in the AA extract groups. Also, in the chromosome aberration test, the AA extract did not cause chromosomal aberration with or without metabolic activation by S9 mix. A bone marrow micronucleus test of mice demonstrated that the incidence of micronucleated polychromatic erythrocytes in the AA extract groups (500, 1000 and 2000 mg/kg BW) was equivalent to that of the negative control group. Based on these results from a standard battery of assays, the AA extract was concluded to have no genotoxic at the proper dose.

References

  1. Park M, Sim M, Kim YC. Anti-Photoaging Effects of Angelica acutiloba Root Ethanol Extract in Human Dermal Fibroblasts. Toxicol Res 2017; 33(2): 125–134.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Sheng Y, Li L, Wang Q, Guo H, Guo DA. Simultaneous determination of gallic acid, albiflorin, paeoniflorin, ferulic acid and benzoic acid in Si-Wu decoction by high-performance liquid chromatography DAD method. J Pharm Biomed Anal 2005; 37(4): 805–810.

    Article  CAS  PubMed  Google Scholar 

  3. Wen K, Huang C, Lu FL. Determination of baicalin and puerarin in traditional Chinese medicinal preparations by high- performance liquid chromatography. J Chromatogr A 1993; 631: 241–250.

    Article  CAS  Google Scholar 

  4. Liu I, Tzeng T, Liou S, Chang CJ. Angelica acutiloba root attenuates insulin resistance induced by high-fructose diet in rats. Phytother Res 2011; 25(9): 1283–1293.

    PubMed  Google Scholar 

  5. Huntley A, Ernst E. A systematic review of herbal medicinal products for the treatment of menopausal symptoms. Menopause 2003; 10(5): 465–476.

    PubMed  Google Scholar 

  6. Jeong S, Kim H, Lee K, Kim K, Huang D, Kim J, Seong RS. Quantitative analysis of marker compounds in Angelica gigas, Angelica sinensis, and Angelica acutiloba by HPLC/DAD. Chem Pharm Bull (Tokyo) 2015; 63(7): 504–511.

    Article  Google Scholar 

  7. Yamada H, Komiyama K, Kiyohara H, Cyong J, Hirakawa Y, Otsuka Y. Structural characterization and antitumor activity of a pectic polysaccharide from the roots of Angelica acutiloba. Planta Med 1990; 56(2): 182–186.

    Article  CAS  PubMed  Google Scholar 

  8. Yoon T, Cheon M, Lee D, Moon B, Lee H, Choo B, Kim HK. Effects of root extracts from Angelica gigas and Angelica acutiloba on inflammatory mediators in mouse macrophages. J Appl Biol Chem 2007; 50(4): 264–269.

    Google Scholar 

  9. Tanaka S, Kano Y, Tabata M, Konoshima M. [Effects of “Toki” (Angelica acutiloba Kitagawa) extracts on writhing and capillary permeability in mice (analgesic and antiinflammatory effects)]. Yakugaku Zasshi 1971; 91(10): 1098–1104.

    Article  CAS  PubMed  Google Scholar 

  10. Tanaka S, Ikeshiro Y, Tabata M, Konoshima M. Anti-nociceptive substances from the roots of Angelica acutiloba. Arzneimittelforschung 1977; 27(11): 2039–2045.

    CAS  PubMed  Google Scholar 

  11. Lee K, Sohn Y, Lee ML Cho H, Jang M, Han N, Shin K, Kim S, Cho I, Bu Y, Jung HS. Effects of Angelica acutiloba on mast cell-mediated allergic reactions in vitro and in vivo. Immunopharmacol Immunotoxicol 2012; 34(4): 571–577.

    Article  PubMed  Google Scholar 

  12. Kumazawa Y, Nakatsuru Y, Fujisawa H, Nishimura C, Mizunoe K, Otsuka Y, Nomoto K. Lymphocyte activation by a polysaccharide fraction separated from hot water extracts of Angelica acutiloba Kitagawa. J Pharmacobiodyn 1985; 8(6): 417–424.

    Article  CAS  PubMed  Google Scholar 

  13. Kiyohara H, Yamada H, Cyong J, Otsuka Y. Studies on polysaccharides from Angelica acutiloba. V. Molecular aggregation and anti-complementary activity of arabinogalactan from Angelica acutiloba. J Pharmacobiodyn 1986; 9(4): 339–346.

    Article  CAS  PubMed  Google Scholar 

  14. Miyazawa M, Tsukamoto T, Anzai J, Ishikawa Y. Insecticidal effect of phthalides and furanocoumarins from Angelica acutiloba against Drosophila melanogaster. J Agric Food Chem 2004; 52(14): 4401–4405.

    Article  CAS  PubMed  Google Scholar 

  15. Liu I, Tzeng T, Liou S, Chang CJ. Regulation of obesity and lipid disorders by extracts from Angelica acutiloba root in high-fat diet-induced obese rats. Phytother Res 2012; 26(2): 223–230.

    Article  CAS  PubMed  Google Scholar 

  16. Lee K, Sohn Y, Lee M, Cho H, Jang M, Han N, Shin K, Kim S, Cho I, Bu Y, Jung HS. Effects of Angelica acutiloba on mast cell-mediated allergic reactions in vitro and in vivo. Immunopharmacol Immunotoxicol 2012; 34(4): 571–577.

    Article  PubMed  Google Scholar 

  17. Joo S, Park D, Shin S, Jeon J, Kim T, Choi Y, Lee S, Kim J, Park S, Hwang B, Lee D, Kim YB. Anti-allergic effects and mechanisms of action of the ethanolic extract of Angelica gigas in dinitrofluorobenzene-induced inflammation models. Environ Toxicol Pharmacol 2010; 30(2): 127–133.

    Article  CAS  PubMed  Google Scholar 

  18. Sarker S, Nahar L. Natural medicine: the genus Angelica. Curr Med Chem 2004; 11(11): 1479–1500.

    Article  CAS  PubMed  Google Scholar 

  19. Uto T, Tung N, Taniyama R, Miyanowaki T, Morinaga O, Shoyama Y Anti-inflammatory Activity of Constituents Isolated from Aerial Part of Angelica acutiloba Kitagawa. Phytother Res 2015; 29(12): 1956–1963.

    Article  CAS  PubMed  Google Scholar 

  20. Markman M. Safety issues in using complementary and alternative medicine. J Clin Oncol 2002; 20: 39–41.

    Article  Google Scholar 

  21. Shin S, Koo K, Bae J, Cha S, Kang I, Kang M, Kim H, Heo H, Park M, Gil G, Lee J, Kim K, Li Y, Lee H, Song S, Choi H, Kang B, Kim JC. Single and 90-day repeated oral dose toxicity studies of fermented Rhus verniciflua stem bark extract in Sprague-Dawley rats. Food Chem Toxicol 2013; 55: 617–626.

    Article  CAS  PubMed  Google Scholar 

  22. Che J, Yun J, Cho E, Kim S, Kim Y, Kim W, Park J, Son W, Kim M, Kang BC. Toxicologic assessment of Paecilomyces tenuipes in rats: renal toxicity and mutagenic potential. Regul Toxicol Pharmacol 2014; 70(2): 527–534.

    Article  CAS  PubMed  Google Scholar 

  23. Che J, Yun J, Kim Y, Kim S, You J, Jang J, Kim H, Kim H, Kang BC. Genotoxicity and subchronic toxicity of Sophorae radix in rats: hepatotoxic and genotoxic potential. Regul Toxicol Pharmacol 2015; 71(3): 379–387.

    Article  CAS  PubMed  Google Scholar 

  24. Yun J, Kim S, Kim Y, You J, Kwon E, Jang J, Park I, Kim H, Kim H, Che J, Kang BC. Evaluation of subchronic (13week) toxicity and genotoxicity potential of vinegar-processed Genkwa Flos. Regul Toxicol Pharmacol 2015; 72(2): 386–393.

    Article  CAS  PubMed  Google Scholar 

  25. Yun J, Che J, Kwon E, Kim Y, Kim S, You J, Kim W, Kim H, Kang BC. Safety evaluation of Angelica gigas: Genotoxicity and 13-weeks oral subchronic toxicity in rats. Regul Toxicol Pharmacol 2015; 72(3): 473–480.

    Article  CAS  PubMed  Google Scholar 

  26. Ministry of Food and Drug Safety (MFDS), 2014. Good Laboratory Practice Regulation for Non-clinical Laboratory Studies (Notification No. 2014-67). MFDS, Korea. Available [October 2014]. https://doi.org/www.mfds.go.kr/ai]27. OECD, 1997. OECD guideline for testing of chemicals, Test No. 471: Bacterial Reverse Mutation test.

    Google Scholar 

  27. OECD, 1997. OECD guideline for testing of chemicals, Test No. 473: in vitro mammalian chromosome aberration test.

    Google Scholar 

  28. OECD, 1997. OECD guideline for testing of chemicals, Test No. 474: mammalian erythrocyte micronucleus test.

    Google Scholar 

  29. Heddle J, Stuart E, Salamone MF. 1984. The bone marrow micronucleus test in Handbook of mutagenicity test procedures. In: Kilbey, B.J., Legator, M., Nichols, W, Ramel, C. (eds.). Elsevier, Amsterdam; pp 441–457.

    Google Scholar 

  30. Hakura A, Suzuki S, Satoh T. Advantage of the use of human liver S9 in the Ames test. Mutat Res 1999; 438(1): 29–36.

    Article  CAS  PubMed  Google Scholar 

  31. Maron D, Ames BN. Revised methods for the Salmonella mutagenicity test. Mutat Res 1983; 113(3-4): 173–215.

    Article  CAS  PubMed  Google Scholar 

  32. Kaleeswaran S, Sriram P, Prabhu D; Chinnathambi; Vijayakumar, Mathuram LN. Anti- and pro-mutagenic effects of silymarin in the Ames bacterial reverse mutation assay. Phytother Res 2009; 23(10): 1378–1384.

    Article  CAS  PubMed  Google Scholar 

  33. Fenech M, Morley AA. Measurement of micronuclei in lymphocytes. Mutat Res 1985; 147(1-2): 29–36.

    Article  CAS  PubMed  Google Scholar 

  34. Perry P, Evans HJ. Cytological detection of mutagen-carcinogen exposure by sister chromatid exchange. Nature 1975; 258(5531): 121–125.

    Article  CAS  PubMed  Google Scholar 

  35. Tucker J, Auletta A, Cimino M, Dearfield K, Jacobson-Kram D, Tice R, Carrano AV. Sister-chromatid exchange: second report of the Gene-Tox Program. Mutat Res 1993; 297(2): 101–108.

    Article  CAS  PubMed  Google Scholar 

  36. Luzhna L, Kathiria P, Kovalchuk O. Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond. Front Genet 2013; 4: 131.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Akyil D, Konuk M. Detection of genotoxicity and mutagenicity of chlorthiophos using micronucleus, chromosome aberration, sister chromatid exchange, and Ames tests. Environ Toxicol 2015; 30(8): 937–945.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Jeong-Hwan Che or Byeong-Cheol Kang.

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Yun, JW., Kim, YS., Kwon, E. et al. Evaluation of in vitro and in vivo genotoxicity of Angelica acutiloba in a standard battery of assays. Lab Anim Res 33, 231–236 (2017). https://doi.org/10.5625/lar.2017.33.3.231

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  • DOI: https://doi.org/10.5625/lar.2017.33.3.231

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