Abstract
Polycyclic aromatic hydrocarbons (PAHs) are interesting environmental pollutants for understanding cocktail effects. High-molecular-weight-PAHs (HMW–PAHs) are classified as probable or possible carcinogens; only benzo[a]pyrene (B[a]P) is a certain carcinogen in humans. Their toxicity depends on their metabolic activation. While 3-hydroxybenzo[a]pyrene (3-OHB[a]P) represents its detoxification pathway, trans-anti-7,8,9,10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene (tetrol-B[a]P) represents the carcinogenicity pathway. The objective was to study the metabolism of B[a]P and HMW–PAHs during chronic low-dose exposure to B[a]P or a PAH mixture. Rats were exposed orally 5 times/week for 10 weeks to low-levels of B[a]P (0.02 and 0.2 mg.kg−1.d−1) or to an industrial mixture extracted from coal tar pitch (CTP) adjusted to 0.2 mg.kg−1.d−1 B[a]P. Urinary levels of monohydroxy-, diol-, and tetrol-PAH were measured during weeks 1 and 10 by HPLC-fluorescence and GC‒MS/MS. After 1 week, the percentages of B[a]P eliminated as 3-OHB[a]P and tetrol-B[a]P were not different depending on the dose of B[a]P, whereas they were reduced by half in the CTP group. Repeated exposure led to an increase in the percentages of the 2 metabolites for the 0.02-B[a]P group. Moreover, the percentage of B[a]P eliminated as 3-OHB[a]P was equal in the 0.2-B[a]P and CTP groups, whereas it remained halved for tetrol-B[a]P in the CTP group. The percent elimination of HMW–PAH metabolites did not vary between weeks 1 and 10. Thus, dose, duration of exposure and chemical composition of the mixture have a major influence on PAH metabolism that goes beyond a simple additive effect. This work contributes to the reflection on determination of limit values and risk assessments in a context of poly-exposures.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Barbeau D, Maitre A, Marques M (2011) Highly sensitive routine method for urinary 3-hydroxybenzo[a]pyrene quantitation using liquid chromatography-fluorescence detection and automated off-line solid phase extraction. Analyst 136(6):1183–1191. https://doi.org/10.1039/c0an00428f
Barbeau D, Persoons R, Marques M, Hervé C, Laffitte-Rigaud G, Maitre A (2014) Relevance of urinary 3-hydroxybenzo(a)pyrene and 1-hydroxypyrene to assess exposure to carcinogenic polycyclic aromatic hydrocarbon mixtures in metallurgy workers. Ann Occup Hyg 58(5):579–590. https://doi.org/10.1093/annhyg/meu004
Barbeau D, Lutier S, Bonneterre V, Persoons R, Marques M, Herve C, Maitre A (2015) Occupational exposure to polycyclic aromatic hydrocarbons: relations between atmospheric mixtures, urinary metabolites and sampling times. Int Arch Occup Environ Health 88(8):1119–1129. https://doi.org/10.1007/s00420-015-1042-1
Barbeau D, Lutier S, Choisinard L, Marques M, Persoons R, Maitre A (2018) Urinary trans-anti-7,8,9,10-tetrahydroxy-7,8,9,10-tetrahydroxybenzo(a)pyrene as the most relevant biomarker for assessing carcinogenic polycyclic aromatic hydrocarbons exposure. Environ Int 112:147–155. https://doi.org/10.1016/j.envint.2017.12.012
Bouchard M, Viau C (1995) Benzo(a)pyrenediolepoxide hemoglobin adducts and 3-hydroxybenzo(a)pyrene urinary excretion profiles in rats subchronically exposed to benzo(a)pyrene. Arch Toxicol 69(8):540–546. https://doi.org/10.1007/s002040050209
Bouchard M, Krishnan K, Viau C (1998) Urinary excretion kinetics of 1-hydroxypyrene following intravenous administration of binary and ternary mixtures of polycyclic aromatic hydrocarbons in rat. Arch Toxicol 72(8):475–482. https://doi.org/10.1007/s002040050531
Bouchard M, Thuot R, Carrier G, Viau C (2002) Urinary excretion kinetics of 1-hydroxypyrene in rats subchronically exposed to pyrene or polycyclic aromatic hydrocarbon mixtures. J Toxicol Environ Health A 65(16):1195–1209. https://doi.org/10.1080/152873902760125408
Bourgart E, Barbeau D, Marques M, von Koschembahr A, Béal D, Persoons R, Leccia MT, Douki T, Maitre A (2019a) A realistic human skin model to study benzo[a]pyrene cutaneous absorption in order to determine the most relevant biomarker for carcinogenic exposure. Arch Toxicol 93(1):81–93. https://doi.org/10.1007/s00204-018-2329-2
Bourgart E, Persoons R, Marques M, Rivier A, Balducci F, von Koschembahr A, Beal D, Leccia MT, Douki T, Maitre A (2019b) Influence of exposure dose, complex mixture, and ultraviolet radiation on skin absorption and bioactivation of polycyclic aromatic hydrocarbons ex vivo. Arch Toxicol 93(8):2165–2184. https://doi.org/10.1007/s00204-019-02504-8
Cooper CS, Ribeiro O, Hewer A, Walsh C, Pal K, Grover PL, Sims P (1980) The involvement of a “bay-region” and a non-’bay-region’ diol-epoxide in the metabolic activation of benz[a]anthracene in mouse skin and in hamster embryo cells. Carcinogenesis 1(3):233–243. https://doi.org/10.1093/carcin/1.3.233
Cornet M, Callaerts A, Jorritsma U, Bolt H, Vercruysse A, Rogiers V (1995) Species-dependent differences in biotransformation pathways of 2-methylpropene (isobutene). Chem Res Toxicol 8(7):987–992. https://doi.org/10.1021/tx00049a013
Dankovic DA, Wright CW, Zangar RC, Springer DL (1989) Complex mixture effects on the dermal absorption of benzo[a]pyrene and other polycyclic aromatic hydrocarbons from mouse skin. J Appl Toxicol 9(4):239–244. https://doi.org/10.1002/jat.2550090407
Gendre C, Lafontaine M, Morele Y, Payan JP, Simon P (2002) Relationship between urinary levels of 1-hydroxypyrene and 3-hydroxybenzo[a]pyrene for workers exposed to polycyclic aromatic hydrocarbons. Polycycl Aromat Compd 22(3–4):761–769. https://doi.org/10.1080/10406630290103915
Godschalk RWL, Moonen EJC, Schilderman PA, Broekmans WM, Kleinjans JC, Van Schooten FJ (2000) Exposure-route-dependent DNA adduct formation by polycyclic aromatic hydrocarbons. Carcinogenesis 21(1):87–92. https://doi.org/10.1093/carcin/21.1.67
Grimmer G, Brune H, Dettbarn G, Heinrich U, Jacob J, Mohtashamipur E, Norpoth K, Pott F, Wenzel-Hartung R (1988) Urinary and faecal excretion of chrysene and chrysene metabolites by rats after oral, intraperitoneal, intratracheal or intrapulmonary application. Arch Toxicol 62(6):401–405. https://doi.org/10.1007/BF00288341
Grimmer G, Jacob J, Dettbarn G, Naujack KW (1997) Determination of urinary metabolites of polycyclic aromatic hydrocarbons (PAH) for the risk assessment of PAH-exposed workers. Int Arch Occup Environ Health 69(4):231–239. https://doi.org/10.1007/s004200050141
Grova N, Faÿs F, Hardy EM, Appenzeller BMR (2017) New insights into urine-based assessment of polycyclic aromatic hydrocarbon-exposure from a rat model: Identification of relevant metabolites and influence of elimination kinetics. Environ Pollut 228:484–495. https://doi.org/10.1016/j.envpol.2017.03.060
Hecht SS, Carmella SG, Villalta PW, Hochalter JB (2010) Analysis of phenanthrene and benzo[a]pyrene tetraol enantiomers in human urine: relevance to the bay region diol epoxide hypothesis of benzo[a]pyrene carcinogenesis and to biomarker studies. Chem Res Toxicol 23(5):900–908. https://doi.org/10.1021/tx9004538
Hilton DC, Trinidad DA, Hubbard K, Li Z, Sjödin A (2017) Measurement of urinary benzo[a]pyrene tetrols and their relationship to other polycyclic aromatic hydrocarbon metabolites and cotinine in humans. Chemosphere 189:365–372. https://doi.org/10.1016/j.chemosphere.2017.09.077
IARC (2010) Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures, IARC monographs on the evaluation of carcinogenic risks to humans, volume 92. Lyon: International Agency for Research on Cancer. ISBN 978-92-832-1292-8
Jongeneelen FJ, Leijdekkers CM, Henderson PT (1984) Urinary excretion of 3-hydroxy-benzo[a]pyrene after percutaneous penetration and oral absorption of benzo[a]pyrene in rats. Cancer Lett 25(2):195–201. https://doi.org/10.1016/s0304-3835(84)80045-2
Jongeneelen FJ, Leijdekkers CM, Bos RP, Theuws JL, Henderson PT (1985) Excretion of 3-hydroxybenzo(a)pyrene and mutagenicity in rat urine after exposure to benzo(a)pyrene. J Appl Toxicol 5(5):277–282. https://doi.org/10.1002/jat.2550050503
Jongeneelen FJ, Anzion RB, Henderson PT (1987) Determination of hydroxylated metabolites of polycyclic aromatic hydrocarbons in urine. J Chromatogr B 413:227–232. https://doi.org/10.1016/0378-4347(87)80230-x
Kang HG, Jeong SH, Cho MH, Cho JH (2007) Changes of biomarkers with oral exposure to benzo(a)pyrene, phenanthrene and pyrene in rats. J Vet Sci 8(4):361–368. https://doi.org/10.4142/jvs.2007.8.4.361
Lafontaine M, Gendre C, Delsaut P, Simon P (2004) Urinary 3-hydroxybenzo[a]pyrene as a biomarker of exposure to polycyclic aromatic hydrocarbons: an approach for determining a biological limit value. Polycycl Aromat Compd 24(4–5):441–450. https://doi.org/10.1080/10406630490471447
Leroyer A, Jeandel F, Maitre A, Howsam M, Deplanque D, Mazzuca M, Nisse C (2010) 1-Hydroxypyrene and 3-hydroxybenzo[a]pyrene as biomarkers of exposure to PAH in various environmental exposure situations. Sci Total Environ 408(5):1166–1173. https://doi.org/10.1016/j.scitotenv.2009.10.073
Levin W, Wood AW, Chang RL, Chang RL, Yagi H, Mah HD, Jerina DM, Conney AH (1978) Evidence for bay region activation of chrysene 1,2-dihydrodiol to an ultimate carcinogen. Cancer Res 38(6):1831–1834
Maitre A, Petit P, Marques M, Hervé C, Montlevier S, Persoons R, Bicout DJ (2018) Exporisq-HAP database: 20 years of monitoring French occupational exposure to polycyclic aromatic hydrocarbon mixtures and identification of exposure determinants. Int J Hyg Environ Health 221(2):334–346. https://doi.org/10.1016/j.ijheh.2017.12.008
Marie C, Bouchard M, Heredia-Ortiz R, Viau C, Maître A (2010) A toxicokinetic study to elucidate 3-hydroxybenzo(a)pyrene atypical urinary excretion profile following intravenous injection of benzo(a)pyrene in rats. J Appl Toxicol 30(5):402–410. https://doi.org/10.1002/jat.1511
Marques M, Maitre A, Choisnard L, Demeilliers C, Persoons R (2021) Simultaneous analysis of PAH urinary mono- and dihydroxylated metabolites by GC-MS-MS following SPE and two-stage derivatization. Anal Bioanal Chem 413(27):6823–6835. https://doi.org/10.1007/s00216-021-03638-4
Miller GW, Jones DP (2014) The nature of nurture: refining the definition of the exposome. Toxicol Sci 137(1):1–2. https://doi.org/10.1093/toxsci/kft251
Moreau M, Bouchard M (2015) Comparison of the kinetics of various biomarkers of benzo[a]pyrene exposure following different routes of entry in rats. J Appl Toxicol 35(7):781–790. https://doi.org/10.1002/jat.3070
Moreau M, Ayotte P, Bouchard M (2015) Kinetics of diol and hydroxybenzo[a]pyrene metabolites in relation to DNA adduct formation and gene expression in rats. J Toxicol Environ Health A 78(12):725–746. https://doi.org/10.1080/15287394.2015.1028119
Pushparajah DS, Umachandran M, Plant KE, Plant N, Ioannides C (2008) Differential response of human and rat epoxide hydrolase to polycyclic aromatic hydrocarbon exposure: studies using precision-cut tissue slices. Mutat Res 640(1–2):153–161. https://doi.org/10.1016/j.mrfmmm.2008.01.004
Ramesh A, Inyang F, Hood DB, Archibong AE, Knuckles ME, Nyanda AM (2001) Metabolism, bioavailability, and toxicokinetics of benzo(α)pyrene in F-344 rats following oral administration. Exp Toxicol Pathol 53(4):275–290. https://doi.org/10.1078/0940-2993-00192
Sen B, Mahadevan B, DeMarini DM (2007) Transcriptional responses to complex mixtures: a review. Mutat Res 636(1–3):144–177. https://doi.org/10.1016/j.mrrev.2007.08.002
Shimada T, Fujii-Kuriyama Y (2004) Metabolic activation of polycyclic aromatic hydrocarbons to carcinogens by cytochromes P450 1A1 and 1B1. Cancer Sci 95(1):1–6. https://doi.org/10.1111/j.1349-7006.2004.tb03162.x
Shimada T, Sugie A, Yamada T, Kawazoe H, Hashimoto M, Azuma E, Nakajima T, Inoue K, Oda Y (2003) Dose-response studies on the induction of liver cytochromes P4501A1 and 1B1 by polycyclic aromatic hydrocarbons in arylhydrocarbon-responsive C57BL/6J mice. Xenobiotica 33(9):957–971. https://doi.org/10.1080/0049825031000140896
Valière M, Petit P, Persoons R, Demeilliers C, Maître A (2022) Consistency between air and biological monitoring for assessing polycyclic aromatic hydrocarbon exposure and cancer risk of workers. Environ Res 207:112268. https://doi.org/10.1016/j.envres.2021.112268
Vineis P, Chadeau-Hyam M, Gmuender H, Gulliver J, Herceg Z, Kleinjans J, Kogevinas M, Kyrtopoulos S, Nieuwenhuijsen M, Phillips DH, Probst-Hensch N, Scalbert A, Vermeulen R, Wild CP, EXPOsOMICS Consortium (2017) The exposome in practice: Design of the EXPOsOMICS project. Int J Hyg Environ Health 220(2):142–151. https://doi.org/10.1016/j.ijheh.2016.08.001
Wild CP (2012) The exposome: from concept to utility. Int J Epidemiol 41(1):24–32. https://doi.org/10.1093/ije/dyr236
Woudneh MB, Benskin JP, Grace R, Hamilton MC, Magee BH, Hoeger GC, Forsberg ND, Cosgrove JR (2016) Quantitative determination of hydroxypolycylic aromatic hydrocarbons as a biomarker of exposure to carcinogenic polycyclic aromatic hydrocarbons. J Chromatogr A 1454:93–100. https://doi.org/10.1016/j.chroma.2016.05.057
Xue W, Warshawsky D (2005) Metabolic activation of polycyclic and heterocyclic aromatic hydrocarbons and DNA damage: a review. Toxicol Appl Pharmacol 206(1):73–93. https://doi.org/10.1016/j.taap.2004.11.006
Zhong Y, Carmella SG, Hochalter JB, Balbo S, Hecht SS (2011) Analysis of r-7, t-8,9, c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene in human urine: a biomarker for directly assessing carcinogenic polycyclic aromatic hydrocarbon exposure plus metabolic activation. Chem Res Toxicol 24(1):73–80. https://doi.org/10.1021/tx100287n
Acknowledgements
We would like to thank the zootechnicians of the « Plateforme de Haute Technologie Animale » (PHTA) facility for animal housing and care, Sylvette Liaudy for her help in the bibliographic research and Franck Balducci for his help in statistical analysis.
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This work was funded by the Translational Innovation in Medicine and Complexity (TIMC) CNRS 5525 laboratory and by the « Ministère de l’enseignement supérieur, de la recherche et de l’innovation» (thesis grant of Maguy El Hajjar). These funders had no role in study design, in the collection, analysis or interpretation of data, in the writing of the report, and in the decision to submit the article for publication.
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Conceived of designed study: CD and AM. Performed animal study: CD, MEH, and MM. Analyzed data: CD, MEH, AM, and RP. Wrote the paper: CD, MEH, AM, MM, and RP.
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Animal housing and procedures were conducted in accordance with the recommendations from the Direction des Services Vétérinaires, Ministry of Agriculture of France, according to European Communities Council Directive 2010/63/EU and according to recommendations for health monitoring from the Federation of European Laboratory Animal Science Associations. Protocols involving animals were reviewed by the local ethics committee « Comité d’Ethique pour l’Expérimentation Animale no.#12, Cometh-Grenoble» and approved by the Ministry of Research (APAFIS#16910-2018092816061560v2).
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El Hajjar, M., Maître, A., Marques, M. et al. Metabolism of benzo[a]pyrene after low-dose subchronic exposure to an industrial mixture of carcinogenic polycyclic aromatic hydrocarbons in rats: a cocktail effect study. Arch Toxicol 97, 865–874 (2023). https://doi.org/10.1007/s00204-023-03441-3
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DOI: https://doi.org/10.1007/s00204-023-03441-3