Environmental Analysis Health and Toxicology

The Korean Society of Environmental Health and Toxicology (환경독성보건학회)
권호 표지

Chemopreventive Effect of Vegetable or Fruit Extract Against Total Diesel Exhaust Particle Extract in NIH/3T3 Cells Using Alkaline Single Cell Gel Electrophoresis

총 디젤분진의 DNA 손상작용과 야채 및 과일추출물의 보호효과

  • Heo Chan (College of Pharmacy, Kangwon National University) ;
  • Kim Nam-Yee (College of Pharmacy, Kangwon National University) ;
  • Heo Moon-Young (College of Pharmacy, Kangwon National University)
  • 허찬 (강원대학교 약학대학) ;
  • 김남이 (강원대학교 약학대학) ;
  • 허문영 (강원대학교 약학대학)
  • Published : 2006.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

In urban areas, diesel exhaust particles (DEP) are probably a major component of particulate matters, especially in Korea where drive many diesel vehicles. The aim of this study was to investigate genotoxic effects of DEP using single ceil gel electrophoresis. In order to evaluate the mechanisms of DEP genotoxicity, the rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed. Total diesel particles (DEPT) was collected without site fractionation from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEPT revealed DNA damage itself in NIH/3T3 cells. The level of DNA breaks plus oxidative DNA lesions and microsome mediated DNA damage was assessed by modified single cell gel eletrophoresis. DEPT was able to induce oxidative DNA damage as well as microsome mediated DNA damage. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor. reduced DNA damage in the presence of S-9 mixture. $DEP_T$ is the sources of oxidative stress, but antioxidants can significantly reduce oxidative DNA dmage. And $DEP_T$ may contain indirect mutagens which can be inhibited by CYP1A1 inhibitors. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) were evaluated for their in vitro antigenotoxic effects. BV and BF showed potent Inhibitory effects against DEPT induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P4501A1 in cell culture.

Keywords

References

  1. 이승철, 허 찬, 이승현, 김현표, 허문영. 야채 및 과일추출물의 항산화작용과 산화적 염색체손상에 대한 억제효과, 약학회지 2004; 48(2): 111-116
  2. 허 찬, 김남이, 김현표, 허문영. 야채 및 과일추출물의 항산화작용, 약학회지 2005; 49(3)
  3. 허 찬, 김남이, 정규혁, 문창규, 허문영. 디젤분진이 체세포에서의 DNA손상에 미치는 영향, 환경독성학회지 2004; 19(4): 335-344
  4. 환경부. 환경백서, 2000
  5. Annas A, Brittebo E and Hellman B. Evaluation of benzo (a)pyrene-induced DNA damage in human endothelial cells using alkaline single cell gel electrophoresis, Mutat. Res. 2000; 471(1-2): 145-155
  6. Baulig A, Garlatti M, Bonvallot V, Marchand A, Barouki R, Marano F and Baeza-Squiban A. Involvement of reactive oxygen species in the metabolic pathways triggered by diesel exhaust particles in human airway epithelial cells, Am J Physiol Lung Cell Mol. Physiol. 2003; 285(3): 671-679 https://doi.org/10.1016/S0887-2333(03)00139-5
  7. Baulig A, Sourdeval M, Meyer M, Marano F and Baeza-Squiban A. Biological effects of atmospheric particles on human bronchial epithelial cells. Comparison with diesel exhaust particles, Toxicol In Vitro 2003; 17(5-6): 567-73
  8. Bonvallot V, Baeza-Squiban A, Baulig A, Brulant S, Boland S, Muzeau R, Barouki R and Marano F. Organic compounds from diesel exhaust particles elicit a proinflammatory response in human airway epithelial cells and induce cytochrome P450 1A1 expression, Am. J. Respir, Cell Mol. Biol 2001; 25: 515-521
  9. Cole SP. Rapid chemosensitivity testing of human lung tumor cells using the MTT assay, Cancer Chemother. Pharmacol 1986; 17: 259-263
  10. Das M, Khan WA, Asokan P, Bickers DR and Mukhtar H. Inhibition of polycyclic aromatic hydrocarbon-DNA adduct formation in epidermis and lungs of SENCAR mice by naturally occurring plant phenols, Cancer Res. 1987; 47(3): 767-773 https://doi.org/10.1021/tx000260u
  11. Dellinger B, Pryor WA, Cueto R, Squadrito GL, Hegde V and Deutsch WA. Role of free radicals in the toxicity of airborne fine particulate matter, Chem. Res. Toxicol. 2001; 14: 1371-1377-145
  12. Don Porto Carero A, Hoet PH, Verschaeve L, Schoeters G and Nemery B. Genotoxic effects of carbon black particles, diesel exhaust particles, and urban air particulates and their extracts on a human alveolar epithelial cell line (A549) and a human monocytic cell line (THP-1), Environ. Mol. Mutagen 2001; 37(2): 155-163
  13. Duez P, Dehon G, and Dubois J. Validation of raw data measurements in the comet assay, Talanta 2004; 63(4): 879-886
  14. Dybdahl M, Risom L, Moller P, Autrup H, Wallin H, Vogel U, Bornholdt J, Daneshvar B, Dragsted LO, Weimann A, Poulsen HE and Loft S. DNA adduct formation and oxidative stress in colon and liver of Big Blue rats after dietary exposure to diesel particles, Carcinogenesis 2003; 24(11): 1759-1766
  15. Edenharder R, Krieg H, Kottgen V and Platt KL. Inhibition of clastogenicity of benzo[a]pyrene and of its trans-7, 8-dihydrodiol in mice in vivo by fruits, vegetables, and flavonoids, Mutat. Res. 2003; 537(2): 169-181
  16. Farombi EO, Moller P and Dragsted LO. Ex-vivo and in vitro protective effects of kolaviron against oxygenderived radical-induced DNA damage and oxidative stress in human lymphocytes and rat liver cells, Cell Biol. Toxicol. 2004; 20(2): 71-82
  17. Gelboin HV. Benzo[a]pyrene metabolism, activation and carcinogenesis: role and regulation of mixed-function oxidases and related enzymes, Physiol. Rev. 1980; 60(4): 1107-1166
  18. Greenwell LL, Moreno T, Jones TP and Richards RJ. Particle-induced oxidative damage is ameliorated by pulmonary antioxidants, Free Radic. Biol. Med. 2002; 32 (9): 898-905 https://doi.org/10.1016/S0891-5849(02)00782-7
  19. Halliwell B and Gutteridge JM, Free Radicals in Biology and Medicine, Oxford University Press 1999
  20. Health Effects Institute (HEI). Diesel exhaust: a critical analysis of emissions, exposure and health effects. A Special Report of the Institute's Diesel Working Group, Health Effects Institute. Cambridge, MA. 1995
  21. Health Effects Institute (HEI). Program summary: Research on Diesel Exhaust, Health Effects Institute. Cambridge, MA. 1999
  22. Heo MY, Sohn SJ and Au WW. Anti-genotoxicity of galangin as a cancer chemopreventive agent candidate, Mutation Research 2001; 488: 135-150
  23. Heo MY, Yu KS, Kim KH, Kim HP and Au WW. Anticlastogenic effect of flavonoid against mutagen-induced micronuclei in mice, Mutation Research 1992; 284: 243-249
  24. Herotog MGL, Hollaman PCH and Katan MB. Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands, J. Agric. Food Chem. 1992; 40: 2379 https://doi.org/10.1021/jf00024a011
  25. International Agency for Research on Cancer (IARC). Diesel and gasoline engine exhausts and some nitroarenes. In: Monographs on the Evaluation of Carcinogenic Risks to Humans, World Health Organization IARC, Lyon, France. 1989; 46
  26. International Agency for Research on Cancer. IARC monographs on the evaluation of carcinogenic risks to humans. Diesel and gasoline engine exhausts and some nitroarenes. International Agency for Research on Cancer. IARC Monogr. Eval. Carcinog. Risks Hum. 1989; 46: 1-458 https://doi.org/10.1016/S0013-9351(05)80104-5
  27. Iwai K, Adachi S, Takahashi M, Moller L, Udagawa T, Mizuno S and Sugawara I. Oxidative DNA damages and lung cancer, Env. Res. 2000; 84(3): 255-264 https://doi.org/10.1016/S0304-3835(97)04676-4
  28. Justesen U, Knuthsen P and Leth T. Determination of plant polyphenols in Danish foodstuffs by HPLC-UV and LCMS detection. Cancer Lett. 1997; 114(1-2): 165 https://doi.org/10.1016/S0304-3835(97)04651-X
  29. Justesen U, Knuthsen P and Leth T. Quantitative analysis of flavonols, flavones, and flavanones in fruits, vegetables and beverages by high-performance liquid chromatography with photo-diode array and mass spectrometric detection. J. Chromatogr. 1998; A99(1-2): 101
  30. Kato A, Nagai A and Kagawa J. Morphological changes in rat lung after long-term exposure to diesel emissions, Inhalation Toxicology 2000; 12: 469-490
  31. Kagawa J. Health effects of diesel exhaust emissions-a mixture of air pollutants of worldwide concern, Toxicology 2002; 27: 349-353
  32. Kim KB and Lee BM. Oxidative stress to DNA, protein, and antioxidant enzymes (superoxide dismutase and catalase) in rats treated with benzo(a)pyrene, Cancer Lett. 1997; 113: 205-212
  33. Kunzli N, Kaiser R, Medina S, Studnicka M, Chanel O, Filliger P, Herry M, Horak F, Jr, Puybonnieux-Texier V and Quenel P. Public-health impact of outdoor and traffic-related air pollution: a European assessment, The Lancet 356(9232): 795-801
  34. Lazarova M, Slamenova D. Genotoxic effects of a complex mixture adsorbed onto ambient air particles on human cells in vitro; the effects of Vitamins E and C, Mutat Res. 2004; 557(2): 167-75 https://doi.org/10.1016/j.mrgentox.2003.10.011
  35. Le Marchand L, Murphy SP, Hankin JH, Wilkens LR and Kolonel LN. Intake of flavonoids and lung cancer, J. Natl. Cancer Inst. 2000; 92(2): 154-160
  36. Lewtas J and Gallagher J. Complex mixtures of urban air pollutants: identification and comparative assessment of mutagenic and tumorigenic chemicals and emission sources, IARC Sci. Publ. 1990; 104: 252-260
  37. Li N, Wang M, Oberley TD, Sempf JM and Nel AE. Comparison of the pro-oxidative and proinflammatory effects of organic diesel exhaust particle chemicals in bronchial epithelial cells and macrophages, J. Immunol. 2002; 169(8): 4531-4541
  38. Li N, Sioutas C, Cho A, Schmitz D, Misra C, Sempf J, Wang M, Oberley T, Froines J and Nel A. Ultrafine particulate pollutants induce oxidative stress and mitochondrial damage, Environ. Health Perspect. 2003; 111: 455-460
  39. Matsuoka A, Furuta A, Ozaki M, Fukuhara K and Miyata N. Resveratrol, a naturally occurring polyphenol, induces sister chromatid exchanges in a Chinese hamster lung (CHL) cell line, Mutat. Res. 2001; 494(1-2): 107 https://doi.org/10.3109/08958379609005424
  40. Mauderly JL. Lung overload: the dilemma and opportunities for resolution, Inhalation Toxicology 1996; 8: 1-28 https://doi.org/10.3109/08958379609005424
  41. McClellan RO. Lung cancer in rats from prolonged exposure to high concentrations of carbonaceous particles: implication for human risk assessment, Inhalation Toxicology 1996; 8: 193-226
  42. Moller P, Daneshvar B, Loft S, Wallin H, Poulsen HE, Autrup H, Ravn-Haren G and Dragsted LO. Oxidative DNA damage in vitamin C-supplemented guinea pigs after intratracheal instillation of diesel exhaust particles, Toxicol. Appl. Pharmacol. 2003; 189(1): 39-44 https://doi.org/10.1016/S0021-9673(97)00984-9
  43. Mouly P, Gaydou EM and Auffray A. Simultaneous separation of flavanone glycosides and polymethoxylated flavones in citrus juices using liquid chromatography. J Chromatogr. A. 1998; 800(2): 171
  44. Muller AK, Farombi EO, Moller P, Autrup HN, Vogel U, Wallin H, Dragsted LO, Loft S and Binderup ML. DNA damage in lung after oral exposure to diesel exhaust particles in Big Blue rats, Mutat. Res. 2004; 550(1-2): 123-132
  45. National Institute for Occupational Safety and Health (NIOSH). Carcinogenic Effects of Exposure to Diesel Exhaust. NIOSH Current Intelligence Bulletin 50. DHHS (NIOSH), Centers for Disease Control. Atlanta, GA. 1988
  46. Pelkonen O and Nebert DW. Metabolism of polycyclic aromatic hydrocarbons: etiologic role in carcinogenesis, Pharmacol. Rev. 1982; 34(2): 189-222
  47. Pohjola SK, Lappi M, Honkanen M, Rantanen L and Savela K. DNA binding of polycyclic aromatic hydrocarbons in a human bronchial epithelial cell line treated with diesel and gasoline particulate extracts and benzo[a]pyrene, Mutagenesis 2003; 18(5): 429-438
  48. Quadri SA, Qadri AN, Hahn ME, Mann KK and Sherr DH. The bioflavonoid galangin blocks aryl hydrocarbon receptor activation and polycyclic aromatic hydrocarbon- induced pre-B cell apoptosis, Mol. Pharmacol. 2000; 58(3): 515-525
  49. Scheepers PT, Coggon D, Knudsen LE, Anzion R, Autrup H, Bogovski S, Bos RP, Dahmann D, Farmer P, Martin EA, Micka V, Muzyka V, Neumann HG, Poole J, Schmidt-Ott A, Seiler F, Volf J and Zwirner-Baier I. BIOMarkers for occupational diesel exhaust exposure monitoring (BIOMODEM)--a study in underground mining, Toxicol. Lett. 2002; 134(1-3): 305-317
  50. Schins RP. Mechanisms of genotoxicity of particles and fibers, Inhal. Toxicol. 2002; 14(1): 57-78
  51. Schwarz D, Kisselev P and Roots I. CYP1A1 genotypeselective inhibition of benzo[a]pyrene activation by quercetin. Eur J Cancer 2005; 41(1): 151-8
  52. Singh NP, McCoy MT, Tice RR and Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells, Experimental Cell Research 1988; 175: 184-191
  53. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ and Klenk DC. Measurement of protein using bicinchoninic acid, Anal. Biochem. 1985; 150: 76-85
  54. Thapliyal R and Maru GB. Inhibition of cytochrome P450 isozymes by curcumins in vitro and in vivo, Food Chem Toxicol 2001; 39(6): 541-547 https://doi.org/10.1016/S0278-6915(00)00165-4
  55. U.S. Environmental Protection Agency (USEPA) Health Assessment Document for Diesel Emissions: SAB Review Draft. EPA/600/8-90/057D. National Center for Environmental Assessment, Washington DC. 1999
  56. World Health Organization (WHO). Diesel fuel and exhaust emissions. International Program on Chemical Safety, Environmental Health Criteria 171, Geneva, Switzerland. 1996
  57. Yang CS, Landau JM, Huang MT and Newmark HL. Inhibition of carcinogenesis by dietary polyphenolic compounds, Annu Rev Nutr. 2001; 21: 381-406 https://doi.org/10.1146/annurev.nutr.21.1.381