Skip to main content
NCBI home page
Toxicological Research logoLink to Toxicological Research
. 2008 Jun 1;24(2):151–159. doi: 10.5487/TR.2008.24.2.151

Evaluation of Genotoxicity of Water and Ethanol Extracts from Rhus verniciflua Stokes (RVS)

Ji-Young Kim 18, Se-Wook Oh 28, Daeseok Han 38, Michael Lee 38,✉
PMCID: PMC7006284  PMID: 32038789

Abstract

Rhus verniciflua Stokes (RVS), one of traditional medicinal plants in Asia, was found to have pharmacological activities such as antioxidative and antiapoptotic effects, raising the possibility for the development of a novel class of anti-cancer drugs. Thus, potential genotoxic effects of RVS in three short-term mutagenicity assays were investigated, which included the Ames assay, in vitro Chromosomal aberration test, and the in vivo Micronucleus assay. In Ames test, the addition of RVS water extracts at doses from 313 up to 5000 mg/plate induced an increase more than 2-fold over vehicle control in the number of revertant colonies in TA98 and TA1537 strains for detecting the frame-shift mutagens. The similar increase in reversion frequency was observed after the addition of RVS ethanol extracts. To assess clastogenic effect, in vitro chromosomal aberration test and in vivo micronucleus assay were performed using Chinese hamster lung cells and male ICR mice, respectively. Both water and ethanol extracts from RVS induced significant increases in the number of metaphases with structural aberrations mostly at concentrations showing the cell survival less than 60% as assessed by in vitro CA test. Also, there was a weak but statistically significant increase in number of micronucleated polychromatic erythrocytes (MNPCEs) in mice treated with water extract at 2000 mg/kg while ethanol extracts of RVS at doses of up to 2000 mg/kg did not induce any statistically significant changes in the incidence of MNPCEs. Therefore, our results lead to conclusion that RVS acts as a genotoxic material based on the available in vitro and in vivo results.

Key words: Rhus verniciflua Stokes (RVS), Ames assay, Chromosomal aberration test, Micronucleus assay, Genotoxicity

References

  1. Choi J, Yoon BJ, Han YN, Lee KT, Ha J, Jung HJ, Park HJ. Antirheumatoid arthritis effect of Rhus verniciflua and of the active component, sulfuretin. Planta Med. 2003;69:899–904. doi: 10.1055/s-2003-38483. [DOI] [PubMed] [Google Scholar]
  2. Dean BJ, Danford N. Assays for the detection of chemically-induced chromosome damage in cultured mammalian cells. In: Venitt S, Parry JM, editors. Mutagenicity testing - a practical approach. Oxford. UK: IRL Press Limited; 1984. pp. 187–232. [Google Scholar]
  3. Fan P, Lou H. Effects of polyphenols from grape seeds on oxidative damage to cellular DNA. Mol. Cell. Biochem. 2004;267:67–74. doi: 10.1023/B:MCBI.0000049366.75461.00. [DOI] [PubMed] [Google Scholar]
  4. Galloway SM. Cytotoxicity and chromosome aberrations in vitro: Experience in industry and the case for an upper limit on toxicity in the aberration assay. Environ. Mol. Mutagen. 2000;35:191–201. doi: 10.1002/(SICI)1098-2280(2000)35:3<191::AID-EM6>3.0.CO;2-4. [DOI] [PubMed] [Google Scholar]
  5. Hayashi M, MacGregor JT, Gatehouse DG. In vivo rodent erythrocyte micronucleus assay. II. Some aspects of protocol design including repeated treatments, integration with toxicity testing and automated scoring. Environ. Mol. Mutagen. 2000;35:234–252. doi: 10.1002/(SICI)1098-2280(2000)35:3<234::AID-EM10>3.0.CO;2-L. [DOI] [PubMed] [Google Scholar]
  6. Heddle JA, Stuart E, Salamone MF. The bone marrow micronucleus test. In: Kilbey BJ, Legator M, Nichols W, Rabel C, editors. Handbook of mutagenicity test procedures. 2nd Edition. 1984. pp. 441–457. [Google Scholar]
  7. Henderson L, Cole R, Cole J, Cole H, Aghamohammdi Z, Regan T. Sister-chromatid exchange and micronucleus induction as indicators of genetic damage in maternal and foetal cells. Mutat. Res. 1984;126:47–52. doi: 10.1016/0027-5107(84)90168-4. [DOI] [PubMed] [Google Scholar]
  8. Hong DH, Han SB, Lee CW, Park SH, Jeon YJ, Kim MJ, Kwak SS, Kim HM. Cytotoxicity of urushiols isolated from sap of Korean lacquer tree (Rhus vernicifera Stokes) Arch. Pharm. Res. 1999;22:638–641. doi: 10.1007/BF02975339. [DOI] [PubMed] [Google Scholar]
  9. Hong M-Y, Kim J-Y, Lee Y-M, Lee M. Assessment of sensitivity of photo-chromosomal assay in the prediction of photo-carcinogenicity. J. Toxicol. Pub. Health. 2005;21:99–105. [Google Scholar]
  10. ICH Harmonized Tripartite Guideline . A Standard Battery for Genotoxicity Testing of Pharmaceuticals, S2B. 1997. [Google Scholar]
  11. Ishidate MJ, Sofuni T, Yoshikawa K. Chromosomal aberration tests in vitro as a primary screening tool for environmental mutagens and/or carcinogens. GANN Monograph on Cancer Res. 1981;27:95–107. [Google Scholar]
  12. Jang HS, Kook SH, Son YO, Kim JG, Jeon YM, Jang YS, Choi KC, Kim J, Han SK, Lee KY, Park BK, Cho NP, Lee JC. Flavonoids purified from Rhus verniciflua Stokes actively inhibit cell growth and induce apoptosis in human osteosarcoma cells. Biochim. Biophys. Acta. 2005;1726:309–316. doi: 10.1016/j.bbagen.2005.08.010. [DOI] [PubMed] [Google Scholar]
  13. Johnson MK, Loo G. Effects of epigallocatechin gallate and quercetin on oxidative damage to cellular DNA. Mutat. Res. 2000;459:211–218. doi: 10.1016/S0921-8777(99)00074-9. [DOI] [PubMed] [Google Scholar]
  14. Jung NC. Biological activity of urushiol and flavonoids from Lac tree. 1998. [Google Scholar]
  15. Kim JH, Kim H-P, Jung C-H, Hong MH, Hong M-C, Bae H-S, Lee S-D, Park S-Y, Park J-H, Ko S- G. Inhibition of cell cycle progression via p27Kip1 upregulation and apoptosis induction by an ethanol extract of Rhus verniciflua Stokes in AGS gastric cancer cells. Int. J. Mol. Med. 2006;18:201–208. [PubMed] [Google Scholar]
  16. Kwon J, Hong M-Y, Koh WS, Chung M-K, Lee M. Computerized Image analysis of micronucleated reticulocytes in mouse bone marrow. J. Toxicol. Pub. Health. 2002;18:369–374. [Google Scholar]
  17. Kim TJ. Korea resource plant. Seoul, Korea: Seoul University Press; 1996. pp. 292–297. [Google Scholar]
  18. Krishna G, Hayashi M. In vivo rodent micronucleus assay: protocol, conduct and data interpretation. Mutat. Res. 2000;455:155–166. doi: 10.1016/S0027-5107(00)00117-2. [DOI] [PubMed] [Google Scholar]
  19. Lee JC, Lim KT, Jang YS. Identification of Rhus verniciflua Stokes compounds that exhibit free radical scavenging and anti-apoptotic properties. Biochim. Biophys. Acta. 2002;1570:181–191. doi: 10.1016/S0304-4165(02)00196-4. [DOI] [PubMed] [Google Scholar]
  20. Lim KT, Hu C, Kitt DD. Antioxidant activity of a Rhus verniciflua Stokes ethanol extract. Food Chem. Toxicol. 2001;39:229–237. doi: 10.1016/S0278-6915(00)00135-6. [DOI] [PubMed] [Google Scholar]
  21. Lovell DP, Anderson D, Albanese R, Amphlett GE, Clare G, Ferguson R, Richold M, Papworth DG, Savage JRK. Statistical analysis on in vivo cytogenetic assays. In: Kirkland DJ, editor. Statistical evaluation of mutagenicity test data. Cambridge, U.K.: Cambridge University Press; 1989. pp. 184–232. [Google Scholar]
  22. MacGregor JT, Jurd L. Mutagenicity of plant flavonoids: structural requirements for mutagenic activity in Salmonella typhimurium. Mutat. Res. 1978;54:297–309. doi: 10.1016/0165-1161(78)90020-1. [DOI] [PubMed] [Google Scholar]
  23. Maron DM, Ames BN. Revised methods for the Salmonella mutagenicity test. Mutat. Res. 1983;113:173–215. doi: 10.1016/0165-1161(83)90010-9. [DOI] [PubMed] [Google Scholar]
  24. Miller WC, Thielman NM, Swai N, Cegielski JP, Shao J, Ting D, Mlalasi J, Manyenga D, Lallinger GJ. Delayed-type hypersensitivity testing in Tanzanian adults with HIV infection. JAIDS-J ACQ IMM DEF. 1996;12:303–308. doi: 10.1097/00042560-199607000-00012. [DOI] [PubMed] [Google Scholar]
  25. Muller L, Sofuni T. Appropriate levels of cytotoxicity for genotoxicity tests using mammalian cells in vitro. Environ. Mol. Mutagen. 2000;35:202–205. doi: 10.1002/(SICI)1098-2280(2000)35:3<202::AID-EM7>3.0.CO;2-Q. [DOI] [PubMed] [Google Scholar]
  26. Na C-S, Choi B-R, Choo D-W, Choi W-I, Kim J-B, Kim H-C, Park YI, Dong M-S. Effect of flavoinoid fractions extracted from Rhus veriniciflua Stokes on the reproductive parameters in SD male rats. J. Toxicol. Pub. Health. 2005;21:309–318. [Google Scholar]
  27. Park KY, Jung GO, Lee KT, Choi J, Choi MY, Kim GT, Jung HJ, Park HJ. Antimutagenic activity of flavonoids from the heartwood of Rhus verniciflua. J. Ethnopharmacol. 2004;90:73–79. doi: 10.1016/j.jep.2003.09.043. [DOI] [PubMed] [Google Scholar]
  28. Richardson C, Williams DA, Allen JA, Amphlett G, Chanter DO, Phillips B. Analysis of data from in vitro cytogenetics assays. In: Kirkland DJ, editor. Statistical evaluation of mutagenicity test data. Cambridge, U.K.: Cambridge University Press; 1989. pp. 141–154. [Google Scholar]
  29. Salamone MF, Heddle JA. The bone marrow micronucleus assay: rationale for a revised protocol. Chemical Mutagens: principles and methods for their detection. 1983;8:111–149. doi: 10.1007/978-1-4613-3694-5_4. [DOI] [Google Scholar]
  30. Son YO, Lee KY, Lee JC, Jang HS, Kim JG, Jeon YM, Jang YS. Selective antiproliferative and apoptotic effects of flavonoids purified from Rhus verniciflua Stokes on normal versus transformed hepatic cell lines. Toxicol. Lett. 2005;155:115–125. doi: 10.1016/j.toxlet.2004.09.003. [DOI] [PubMed] [Google Scholar]
  31. Sung B, Pandey MK, Aggarwal BB. Fisetin, an inhibitor of cyclin-dependent kinase 6, down-Regulates NF-κB-regulated cell proliferation, antiapoptotic and metastatic gene products through the suppression of TAK- 1 and RIP regulated IκB-α kinase activation. Mol. Pharmacol. 2007;71:1703–1714. doi: 10.1124/mol.107.034512. [DOI] [PubMed] [Google Scholar]
  32. The Collaborative Study Group for the Micronucleus Test Sex difference in the micronucleus test. Mutat. Res. 1986;172:151–163. doi: 10.1016/0165-1218(86)90071-6. [DOI] [PubMed] [Google Scholar]
  33. Tinwell H, Ashby J. Comparison of acridine orange and Giemsa stains in several mouse bone marrow micronucleus assay - including a triple dose study. Mutagenesis. 1989;4:476–481. doi: 10.1093/mutage/4.6.476. [DOI] [PubMed] [Google Scholar]
  34. Yang EB, Zhang K, Cheng LY, Mack P. Butein, a specific protein tyrosine kinase inhibitor. Biochem. Biophys. Res. Commun. 1998;245:435–438. doi: 10.1006/bbrc.1998.8452. [DOI] [PubMed] [Google Scholar]

Articles from Toxicological Research are provided here courtesy of Korean Society of Toxicology

RESOURCES