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In vitro Antimutagenic and Genotoxic Effects of Azadirachta indica Extract

님추출물의 in vitro 항돌연변이원성 및 유전독성 영향

  • Yoon, Hyunjoo (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Cho, Hyeon-Jo (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Kim, Jin Hyo (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Park, Kyung-Hun (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Gil, Geun-Hwan (Agro-Material Safety Evaluation Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Oh, Jin-Ah (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Cho, Namjun (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration) ;
  • Paik, Min-Kyoung (Chemical Safety Division, National Academy of Agricultural Sciences, Rural Development Administration)
  • Received : 2013.10.21
  • Accepted : 2014.02.07
  • Published : 2014.09.30

Abstract

Azadirachta indica extract (AIE) has been regarded as a promising source of environment-friendly organic materials owing to their low mammalian toxicity. However, quite a bit of research has been reported that AIE may cause clastogens in human lymphocytes. Therefore, this study was conducted to evaluate the antimutagenic and genotoxicity of two samples of AIE. Antimutagenic test was experimented by using bacterial reverse mutation test. In the bacterial reverse mutation test, five strains Salmonella Typhimurim of two samples of AIE in order to evaluate its mutagenic potential. Bacterial reverse mutation test was also performed on positive control and negative control groups in the presence of the metabolic activation system (S-9 mix) and metabolic non-activation system. In the chromosome aberration test, Chinese hamster lung cells were exposed to AIE for 6 or 24 h with BPS, or for 6 h with S-9 mix. Negative and positive control groups were experimented for chromosome aberration test. As a result, the number of mutated colonies induced by 4-NQO were reduced by AIE treatment in all strains, indicating that AIE may have antimutagenic effects. Bacterial reverse mutation and chromosomal aberration were not shown at all concentration of AIE, regardless of activation of the metabolic system. we concluded that two AIE samples used in this study have no genotoxic effects to human, according to the genotoxicity battery system suggested by ICH (International Conference on Harmonization).

님 추출물은 포유류에 낮은 독성을 가지고 있기 때문에 유기농업자재로 주로 이용되고 있다. 그러나 님 추출물은 림프구의 염색체 이상을 야기한다는 연구결과가 보고되었다. 따라서, 본 연구는 님 추출물을 이용하여 항돌연변이원성 시험 및 유전독성을 평가하였다. 항돌연변이원성 시험은 복귀돌연변이 시험을 이용하여 시험하였다. 복귀돌연변이 시험은 2개의 님 추출물을 Salmonella Typhimurium 5개 균주를 이용하여 돌연변이 유발 가능성을 평가하였다. 복귀돌연변이시험과 염색체이상시험은은 대사활성계 처리군(S-9 mix)과 대사활성계 미처리군(PBS)으로 나누어 양성대조군과 음성대조군을 사용하여 실시하였다. 염색체이상시험은 Chinese hamster lung cell을 이용하여 님추출물 시료에 대사활성계 처리군은 6시간 노출시켰고, 대사활성계 미처리군은 각각 6시간과 24시간 노출시켜 시험하였고, 음성대조군과 양성대조군을 사용하였다. 4 NQO에 의해 유도 된 돌연변이 집락수는 님추출물 시료 처리에 의해 감소되어 SRE는 항 돌연변이 효과가 있을 수 있음을 나타냈다. 복귀돌연변이와 염색체이상시험은 님추출물 모든 시험 농도군에서 대화활성계의 처리 유무와 관계없이 음성으로 판정되었다. 이상의 결과를 ICH에서 제안된 유전독성 battery system에 근거해 살펴 봤을 때 본 연구에서 사용된 님 추출물 2종은 모두 유전독성이 없어 안전함을 확인 할 수 있었다.

Keywords

References

  1. Awasthy KS, Chaurasia OP, and Sinha SP (1995) Genotoxic effects of crude extract of neem (Azadirachta indica) in bone marrow cells of mice. Cytologia 60, 189-95. https://doi.org/10.1508/cytologia.60.189
  2. Awasthy KS, Chaurasia OP, and Sinha SP (1999) Prolonged murine genotoxic effects of crude extracted from neem. Phytother Res 11, 81-3.
  3. Boeke SJ, Boersma MG, Alink GM, van Loon JJ, van Huis A, Dicke M et al. (2004) Safety evaluation of neem (azadirachta indica) derived pesticides. J Ethnophamacol 94, 25-41. https://doi.org/10.1016/j.jep.2004.05.011
  4. Cho HY, Yoon HY, Park KY, Lee JB, Shim CK, Kim JH et al. (2013) In vitro antimutagenic and genotoxic effects of Sophora Radis Extracts. Korean J Pesticide Science 17(4), 1-8. https://doi.org/10.7585/kjps.2013.17.1.1
  5. EFSA(European Food Safety Authority) (2011) Conclusion on the peer review of the pesticide risk assessment of the active substance azadirachtin. EFSA Journal 9, 1858 https://doi.org/10.2903/j.efsa.2011.1858
  6. El-Hawary ZM and Kholief TS (1990) Biochemical studies on hypoglycemic agents. I. Effect of Azadirachta indica leaf extract. Arch Pharm Research (Seoul) 13, 108-12. https://doi.org/10.1007/BF02857845
  7. Govindachari TR, Suresh G, Gopalakrishnan G, Banumathy B, and Masilamani S (1998) Identification of antifungal compounds from the seed oil of Azadirachta indica. Phytoparasitica 26, 109-16. https://doi.org/10.1007/BF02980677
  8. Han S (1989) Diversion of locomotor activity rhythm and neural control by Azadirachtin in Leucophaea maderae. Korean J Zool 32, 441-9.
  9. ICH (2012) Genotoxicity testing and data interpretation for pharmaceuticals intended for human use. International Conference on Harmonization, USA.
  10. Ishidate M Jr and Odashima S (1977) Chromosome test with 134 compounds on Chinese hamster cells in vitro-A screening for chemicals carcinogens. Mutat Res 48, 337-54. https://doi.org/10.1016/0027-5107(77)90177-4
  11. JEMS-MMS (1988) Atlas of chromosome aberration by chemicals. Japanese Environmental Mutagen Society-Mammalian Mutagenicity Study Group, Japan.
  12. Jeun J, Cho HJ, Jun H, Lee JH, Jia Y, Cho KS et al. (2011) Mutagenic and antimutagenic effects of hemp seed oil evaluated by ames Salmonella testing. Korean J Food Sci Technol 43, 396-400. https://doi.org/10.9721/KJFST.2011.43.3.396
  13. Jongen WM and Koeman JH (1983) Mutagenicity testing of two tropical plant materials with pesticidal potential in Salmonella typhimurium: Phytolacca dodecandraberries and oil from seeds of Azadirachta indica. Environ Mutagen 5, 687-94. https://doi.org/10.1002/em.2860050507
  14. KEFAM Association (2012) Standard guidelines of environment-friendly organic materials. Korea Eco-friendly Agro-materials, Korea.
  15. Kusamran WR, Tepsuwan A, and Kupradinun P (1998) Antimutagenic and anticarcinogenic potentials of some Thai vegetables. Mutat Res 18, 247-58.
  16. Lee J-W, Jin C-L, Jang KC, Choi G-H, Lee H-D, and Kim JH (2013) Investigation on the insecticidal limonoid content of commercial biopesticides and neem extract using solid phase extraction. J Agric Chem Environ 2(4), 81-5.
  17. Maron DM and Ames BN (1983) Reversed methods for the Salmonella mutagenicity test. Mutat Res 113, 173-215. https://doi.org/10.1016/0165-1161(83)90010-9
  18. Nakahara K, Roy MK, Ono H, Maeda I, Ohnishi-Kameyama M, Yoshida M et al. (2003) Prenylated flavanones isolated from flowers of antimutagenic constituents against heterocyclic amines. J Agric Food Chem 51, 6456-60. https://doi.org/10.1021/jf034666z
  19. OECD SIDS (1999) Acetone, SIDS Initial Assessment Report for the 9th SIAM. UNEP Publications, USA.
  20. Park M, Kim J, Choi W, and Yoon M (2011) Nematicidal effect of root-knot nematode (Meloidogyne incognita) by biological nematicide. Korean J Soil Fert 44, 228-35. https://doi.org/10.7745/KJSSF.2011.44.2.228
  21. Paterson RR (2006) Ganoderma-a therapeutic fungal biofactory. Phytochemistry 67, 1985-2001. https://doi.org/10.1016/j.phytochem.2006.07.004
  22. Polasa K and Rukmini C (1987) Mutagenicity tests of cashew nut shell liquid, rice-bran oil and other vegetable oils using the Salmonella typhimurium/microsome system. Food Chem Toxicol 25, 763-6. https://doi.org/10.1016/0278-6915(87)90231-6
  23. Rojanapo W and Tepsuwan A (1992) Matagenic and antimutagenic activities of some vegetables (in Thai). Bulletin of the Department of Medical Services 17, 461-9.
  24. RosenKranz HS and Klopman G (1995) An examination of the potential genotoxic carcinogenicity of a biopesticide derived from neem tree. Environ Mol Mutagen 26, 255-60. https://doi.org/10.1002/em.2850260311
  25. Schmutterer H (1990) Properties and potential of natural pesticides form the neem tree, Azadirachta indica. Annu Rev Entomol 35, 271-97. https://doi.org/10.1146/annurev.en.35.010190.001415
  26. Shin N and Koo S (1998) Antimutagenic effects of dietary fiber from yam (dioscorea batatas decne) against 2-AF and MNNG. Korean J Soc Food Sci 14, 333-8.
  27. Singh PP, Junnarkar AY, Reddi GS, and Singh KV (1987) Azadirachta indica: neuro-sychopharmacological antimicrobial studies. Fitoterapia 58, 235-8.
  28. Song S, Yang DC, and Choung SY (2005) The chromosomal aberration test of wild ginseng culture extract in chinese hamster lung cell. J Toxicol Pub Health 21, 57-62.
  29. Tandan SK, Gupta S, Chandra S, Lal J, and Singh R (1995) Safety evaluation of Azadirachta indica seed oil, a herbal wound dressing agent. Fitoterapia 66, 69-72.
  30. Vinod V, Tiwari PK, and Meshram GP (2011) Evaluation of mutagenic and antimutagenic activities of neem (Azadirachta indica) seed oil in the in vitro Ames Salmonella/microsome assay and in vivo mouse bone marrow micronucleus test. J Ethnopharmacol 134, 931-7. https://doi.org/10.1016/j.jep.2011.02.003

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