Abstract
Heterocyclic amines (HCAs) are well-known for their mutagenic properties. One of the major routes of human exposure is through consumption of cooked meat, as certain cooking methods favor formation of HCAs. Recent epidemiological studies reported significant associations between dietary HCA exposure and insulin resistance and type II diabetes. However, no previous studies have examined if HCAs, independent of meat consumption, contributes to pathogenesis of insulin resistance or metabolic disease. In the present study, we have assessed the effect of three HCAs commonly found in cooked meat (2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline [MeIQ], 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline [MeIQx], and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine [PhIP]) on insulin signaling and glucose production. HepG2 or cryopreserved human hepatocytes were treated with 0–50 μM of MeIQ, MeIQx, or PhIP for 3 days. Treatment of HepG2 cells and hepatocytes with MeIQ and MeIQx resulted in a significant reduction in insulin-induced AKT phosphorylation, suggesting that HCA exposure decreases hepatic insulin signaling. HCA treatment also led to significant increases in expression of gluconeogenic genes, G6PC and PCK1, in both HepG2 and cryopreserved human hepatocytes. Additionally, the level of phosphorylated FOXO1, a transcriptional regulator of gluconeogenesis, was significantly reduced by HCA treatment in hepatocytes. Importantly, HCA treatment of human hepatocytes led to increases in extracellular glucose level in the presence of gluconeogenic substrates, suggesting that HCAs induce hepatic glucose production. The current findings suggest that HCAs induce insulin resistance and promote hepatic glucose production in human hepatocytes. This implicates that exposure to HCAs may lead to the development of type II diabetes or metabolic syndrome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets for this study are available from the corresponding author upon request.
Change history
04 September 2023
A Correction to this paper has been published: https://doi.org/10.1007/s00204-023-03596-z
References
Ahrens M, Ammerpohl O, von Schönfels W et al (2013) DNA methylation analysis in nonalcoholic fatty liver disease suggests distinct disease-specific and remodeling signatures after bariatric surgery. Cell Metab 18:296–302. https://doi.org/10.1016/j.cmet.2013.07.004
Alves TC, Befroy DE, Kibbey RG et al (2011) Regulation of hepatic fat and glucose oxidation in rats with lipid-induced hepatic insulin resistance. Hepatol Baltim Md 53:1175–1181. https://doi.org/10.1002/hep.24170
Brown MS, Goldstein JL (2008) Selective versus total insulin resistance: a pathogenic paradox. Cell Metab 7:95–96. https://doi.org/10.1016/j.cmet.2007.12.009
Carvalho AM, Miranda AM, Santos FA et al (2015) High intake of heterocyclic amines from meat is associated with oxidative stress. Br J Nutr 113:1301–1307. https://doi.org/10.1017/S0007114515000628
Cheng K-W, Chen F, Wang M (2006) Heterocyclic amines: chemistry and health. Mol Nutr Food Res 50:1150–1170. https://doi.org/10.1002/mnfr.200600086
Chou HC, Lang NP, Kadlubar FF (1995) Metabolic activation of N-hydroxy arylamines and N-hydroxy heterocyclic amines by human sulfotransferase(s). Cancer Res 55:525–529
de Luca C, Olefsky JM (2008) Inflammation and insulin resistance. FEBS Lett 582:97–105. https://doi.org/10.1016/j.febslet.2007.11.057
Eisenbrand G, Tang W (1993) Food-borne heterocyclic amines. Chemistry, formation, occurrence and biological activities. A literature review. Toxicology 84:1–82. https://doi.org/10.1016/0300-483x(93)90109-6
Estall JL, Kahn M, Cooper MP et al (2009) Sensitivity of lipid metabolism and insulin signaling to genetic alterations in hepatic peroxisome proliferator-activated receptor-gamma coactivator-1alpha expression. Diabetes 58:1499–1508. https://doi.org/10.2337/db08-1571
Felton JS, Knize MG, Hatch FT et al (1999) Heterocyclic amine formation and the impact of structure on their mutagenicity. Cancer Lett 143:127–134. https://doi.org/10.1016/s0304-3835(99)00141-x
Goffart S, Wiesner RJ (2003) Regulation and co-ordination of nuclear gene expression during mitochondrial biogenesis. Exp Physiol 88:33–40. https://doi.org/10.1113/eph8802500
Gonzalez E, McGraw TE (2009) Insulin-modulated Akt subcellular localization determines Akt isoform-specific signaling. Proc Natl Acad Sci USA 106:7004–7009. https://doi.org/10.1073/pnas.0901933106
Gross DN, van den Heuvel APJ, Birnbaum MJ (2008) The role of FoxO in the regulation of metabolism. Oncogene 27:2320–2336. https://doi.org/10.1038/onc.2008.25
Gu D, McNaughton L, Lemaster D et al (2010) A comprehensive approach to the profiling of the cooked meat carcinogens 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, and their metabolites in human urine. Chem Res Toxicol 23:788–801. https://doi.org/10.1021/tx900436m
Gual P, Le Marchand-Brustel Y, Tanti J-F (2005) Positive and negative regulation of insulin signaling through IRS-1 phosphorylation. Biochimie 87:99–109. https://doi.org/10.1016/j.biochi.2004.10.019
Guengerich FP (1992) Metabolic activation of carcinogens. Pharmacol Ther 54:17–61. https://doi.org/10.1016/0163-7258(92)90050-a
Han H-S, Kang G, Kim JS et al (2016) Regulation of glucose metabolism from a liver-centric perspective. Exp Mol Med 48:e218. https://doi.org/10.1038/emm.2015.122
Hein DW (2002) Molecular genetics and function of NAT1 and NAT2: role in aromatic amine metabolism and carcinogenesis. Mutat Res 506–507:65–77. https://doi.org/10.1016/s0027-5107(02)00153-7
Hong KU, Salazar-González RA, Walls KM, Hein DW (2022) Transcriptional regulation of human arylamine N-acetyltransferase 2 gene by glucose and insulin in liver cancer cell lines. Toxicol Sci 190:158–172. https://doi.org/10.1093/toxsci/kfac103
Houstis N, Rosen ED, Lander ES (2006) Reactive oxygen species have a causal role in multiple forms of insulin resistance. Nature 440:944–948. https://doi.org/10.1038/nature04634
Kamata H, Honda S-I, Maeda S et al (2005) Reactive oxygen species promote TNFalpha-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell 120:649–661. https://doi.org/10.1016/j.cell.2004.12.041
Kersten S, Stienstra R (2017) The role and regulation of the peroxisome proliferator activated receptor alpha in human liver. Biochimie 136:75–84. https://doi.org/10.1016/j.biochi.2016.12.019
Kimura S, Kawabe M, Yu A et al (2003) Carcinogenesis of the food mutagen PhIP in mice is independent of CYP1A2. Carcinogenesis 24:583–587. https://doi.org/10.1093/carcin/24.3.583
Knasmüller S, Schwab CE, Land SJ et al (1999) Genotoxic effects of heterocyclic aromatic amines in human derived hepatoma (HepG2) cells. Mutagenesis 14:533–540. https://doi.org/10.1093/mutage/14.6.533
Knize MG, Dolbeare FA, Cunningham PL, Felton JS (1995) Mutagenic activity and heterocyclic amine content of the human diet. Princess Takamatsu Symp 23:30–38
Koliaki C, Szendroedi J, Kaul K et al (2015) Adaptation of hepatic mitochondrial function in humans with non-alcoholic fatty liver is lost in steatohepatitis. Cell Metab 21:739–746. https://doi.org/10.1016/j.cmet.2015.04.004
Lan CM, Kao TH, Chen BH (2004) Effects of heating time and antioxidants on the formation of heterocyclic amines in marinated foods. J Chromatogr B Anal Technol Biomed Life Sci 802:27–37. https://doi.org/10.1016/j.jchromb.2003.09.025
Langouët S, Welti DH, Kerriguy N et al (2001) Metabolism of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline in human hepatocytes: 2-amino-3-methylimidazo[4,5-f]quinoxaline-8-carboxylic acid is a major detoxification pathway catalyzed by cytochrome P450 1A2. Chem Res Toxicol 14:211–221. https://doi.org/10.1021/tx000176e
Leone TC, Lehman JJ, Finck BN et al (2005) PGC-1alpha deficiency causes multi-system energy metabolic derangements: muscle dysfunction, abnormal weight control and hepatic steatosis. PLoS Biol 3:e101. https://doi.org/10.1371/journal.pbio.0030101
Lin HV, Accili D (2011) Hormonal regulation of hepatic glucose production in health and disease. Cell Metab 14:9–19. https://doi.org/10.1016/j.cmet.2011.06.003
Liu G, Zong G, Hu FB et al (2017) Cooking methods for red meats and risk of type 2 diabetes: a prospective study of US women. Diabetes Care 40:1041–1049. https://doi.org/10.2337/dc17-0204
Liu G, Zong G, Wu K et al (2018) Meat cooking methods and risk of type 2 diabetes: results from three prospective cohort studies. Diabetes Care 41:1049–1060. https://doi.org/10.2337/dc17-1992
Mackenzie RW, Elliott BT (2014) Akt/PKB activation and insulin signaling: a novel insulin signaling pathway in the treatment of type 2 diabetes. Diabetes Metab Syndr Obes Targets Ther 7:55–64. https://doi.org/10.2147/DMSO.S48260
Metry KJ, Neale JR, Bendaly J et al (2009) Effect of N-acetyltransferase 2 polymorphism on tumor target tissue DNA adduct levels in rapid and slow acetylator congenic rats administered 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine or 2-amino-3,8-dimethylimidazo-[4,5-f]quinoxaline. Drug Metab Dispos Biol Fate Chem 37:2123–2126. https://doi.org/10.1124/dmd.109.029512
Metry KJ, Neale JR, Doll MA et al (2010) Effect of rapid human N-acetyltransferase 2 haplotype on DNA damage and mutagenesis induced by 2-amino-3-methylimidazo-[4,5-f]quinoline (IQ) and 2-amino-3,8-dimethylimidazo-[4,5-f]quinoxaline (MeIQx). Mutat Res 684:66–73. https://doi.org/10.1016/j.mrfmmm.2009.12.001
Morris EM, Meers GME, Booth FW et al (2012) PGC-1α overexpression results in increased hepatic fatty acid oxidation with reduced triacylglycerol accumulation and secretion. Am J Physiol Gastrointest Liver Physiol 303:G979-992. https://doi.org/10.1152/ajpgi.00169.2012
Nebert DW (1991) Role of genetics and drug metabolism in human cancer risk. Mutat Res 247:267–281. https://doi.org/10.1016/0027-5107(91)90022-g
Orzechowski A, Schrenk D, Bock-Hennig BS, Bock KW (1994) Glucuronidation of carcinogenic arylamines and their N-hydroxy derivatives by rat and human phenol UDP-glucuronosyltransferase of the UGT1 gene complex. Carcinogenesis 15:1549–1553. https://doi.org/10.1093/carcin/15.8.1549
Petersen MC, Shulman GI (2018) Mechanisms of insulin action and insulin resistance. Physiol Rev 98:2133–2223. https://doi.org/10.1152/physrev.00063.2017
Puigserver P, Rhee J, Donovan J et al (2003) Insulin-regulated hepatic gluconeogenesis through FOXO1-PGC-1alpha interaction. Nature 423:550–555. https://doi.org/10.1038/nature01667
Roberts CK, Hevener AL, Barnard RJ (2013) Metabolic syndrome and insulin resistance: underlying causes and modification by exercise training. Compr Physiol 3:1–58. https://doi.org/10.1002/cphy.c110062
Saklayen MG (2018) The global epidemic of the metabolic syndrome. Curr Hypertens Rep 20:12. https://doi.org/10.1007/s11906-018-0812-z
Salvadó L, Palomer X, Barroso E, Vázquez-Carrera M (2015) Targeting endoplasmic reticulum stress in insulin resistance. Trends Endocrinol Metab 26:438–448. https://doi.org/10.1016/j.tem.2015.05.007
Sangwung P, Petersen KF, Shulman GI, Knowles JW (2020) Mitochondrial dysfunction, insulin resistance, and potential genetic implications. Endocrinology. https://doi.org/10.1210/endocr/bqaa017
Santoleri D, Titchenell PM (2019) Resolving the paradox of hepatic insulin resistance. Cell Mol Gastroenterol Hepatol 7:447–456. https://doi.org/10.1016/j.jcmgh.2018.10.016
Sefried S, Häring H-U, Weigert C, Eckstein SS (2018) Suitability of hepatocyte cell lines HepG2, AML12 and THLE-2 for investigation of insulin signalling and hepatokine gene expression. Open Biol 8:180147. https://doi.org/10.1098/rsob.180147
Storz P, Toker A (2003) Protein kinase D mediates a stress-induced NF-kappaB activation and survival pathway. EMBO J 22:109–120. https://doi.org/10.1093/emboj/cdg009
Toribio F, Moyano E, Puignou L, Galceran MT (2002) Ion-trap tandem mass spectrometry for the determination of heterocyclic amines in food. J Chromatogr A 948:267–281. https://doi.org/10.1016/s0021-9673(01)01476-5
Turesky RJ (2007) Formation and biochemistry of carcinogenic heterocyclic aromatic amines in cooked meats. Toxicol Lett 168:219–227. https://doi.org/10.1016/j.toxlet.2006.10.018
Westerbacka J, Kolak M, Kiviluoto T et al (2007) Genes involved in fatty acid partitioning and binding, lipolysis, monocyte/macrophage recruitment, and inflammation are overexpressed in the human fatty liver of insulin-resistant subjects. Diabetes 56:2759–2765. https://doi.org/10.2337/db07-0156
Woziwodzka A, Tarasewicz M, Piosik J (2010) Heterocyclic aromatic amines, food-derived mutagens: metabolism and relevance to cancer susceptibility. Postepy Biochem 56:435–446
Xian Y, Wu Y, Dong H et al (2019) Modified QuEChERS purification and Fe3O4 nanoparticle decoloration for robust analysis of 14 heterocyclic aromatic amines and acrylamide in coffee products using UHPLC-MS/MS. Food Chem 285:77–85. https://doi.org/10.1016/j.foodchem.2019.01.132
Xing Y, Li A, Yang Y et al (2018) The regulation of FOXO1 and its role in disease progression. Life Sci 193:124–131. https://doi.org/10.1016/j.lfs.2017.11.030
Yang W-M, Min K-H, Lee W (2016) MiR-1271 upregulated by saturated fatty acid palmitate provokes impaired insulin signaling by repressing INSR and IRS-1 expression in HepG2 cells. Biochem Biophys Res Commun 478:1786–1791. https://doi.org/10.1016/j.bbrc.2016.09.029
Yoon JC, Puigserver P, Chen G et al (2001) Control of hepatic gluconeogenesis through the transcriptional coactivator PGC-1. Nature 413:131–138. https://doi.org/10.1038/35093050
Zamora R, Hidalgo FJ (2020) Formation of heterocyclic aromatic amines with the structure of aminoimidazoazarenes in food products. Food Chem 313:126128. https://doi.org/10.1016/j.foodchem.2019.126128
Zelber-Sagi S, Ivancovsky-Wajcman D, Fliss Isakov N et al (2018) High red and processed meat consumption is associated with non-alcoholic fatty liver disease and insulin resistance. J Hepatol 68:1239–1246. https://doi.org/10.1016/j.jhep.2018.01.015
Zhang W, Patil S, Chauhan B et al (2006) FoxO1 regulates multiple metabolic pathways in the liver: effects on gluconeogenic, glycolytic, and lipogenic gene expression. J Biol Chem 281:10105–10117. https://doi.org/10.1074/jbc.M600272200
Zhang L, Ashley DL, Watson CH (2011) Quantitative analysis of six heterocyclic aromatic amines in mainstream cigarette smoke condensate using isotope dilution liquid chromatography-electrospray ionization tandem mass spectrometry. Nicotine Tob Res 13:120–126. https://doi.org/10.1093/ntr/ntq219
Zhang Z, Liu H, Liu J (2019) Akt activation: a potential strategy to ameliorate insulin resistance. Diabetes Res Clin Pract 156:107092. https://doi.org/10.1016/j.diabres.2017.10.004
Zhang L, Wang L, Li Y et al (2020) Evaluation of tobacco smoke and diet as sources of exposure to two heterocyclic aromatic amines for the US population: NHANES 2013–2014. Cancer Epidemiol Biomark Prev Publ Am Assoc Cancer Res Cosponsored Am Soc Prev Oncol 29:103–111. https://doi.org/10.1158/1055-9965.EPI-19-0169
Acknowledgements
This work was partially supported by United States Public Health Service Grants P20-GM113226, P30-ES030283, T32-ES011564 and P42-ES023716. Portions of this work will constitute partial fulfilment by Kennedy Walls for the Ph.D. in pharmacology and toxicology at the University of Louisville.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Walls, K.M., Hong, K.U. & Hein, D.W. Heterocyclic amines reduce insulin-induced AKT phosphorylation and induce gluconeogenic gene expression in human hepatocytes. Arch Toxicol 97, 1613–1626 (2023). https://doi.org/10.1007/s00204-023-03488-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-023-03488-2