Abstract
Early-life consumption of high-fat and sugar-rich foods is recognized as a major contributor for the onset of metabolic dysfunction and its related disorders, including diabetes and nonalcoholic fatty liver disease. The lifelong impact of early unhealthy eating habits that start at younger ages remains unclear. Therefore, to better understand the effects of diet, it is essential to evaluate the structural and functional changes induced in metabolic organs and potential mechanisms underlying those changes. To investigate the long-term effects of eating habits, young male rats were exposed to high-sugar and high-energy diets. After 14 weeks, body composition was assessed, and histopathological changes were analyzed in the liver and adipose tissue. Serum biochemical parameters were also determined. Expression of inflammatory markers in the liver was evaluated by immunohistochemistry. Our results revealed that serum levels of glucose, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), and lipid profile were increased in rats red high-sugar and high-energy diets. Histopathological alterations were observed, including abnormal hepatocyte organization and lipid droplet accumulation in the liver, and abnormal structure of adipocytes. In both unhealthy diet groups, hepatic expression of Toll-like receptor 4 (TLR4), cyclooxygenase 2 (COX-2), and E-selectin were increased, as well as a biomarker of oxidative stress. Together, our data demonstrated that unhealthy diets induced functional and structural changes in the metabolic organs, suggesting that proinflammatory and oxidative stress mechanisms trigger the hepatic alterations and metabolic dysfunction.
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References
Ainslie DA, Proietto J, Fam BC, Thorburn AW (2000) Short-term, high-fat diets lower circulating leptin concentrations in rats. Am J Clin Nutr 71:438–442. https://doi.org/10.1093/ajcn/71.2.438
Angulo P (2002) Nonalcoholic fatty liver disease. N Engl J Med 346:1221–1231. https://doi.org/10.1056/NEJMra011775
Arriarán S, Agnelli S, Sabater D, Remesar X, Fernández-López JA, Alemany M (2015) Evidences of basal lactate production in the main white adipose tissue sites of rats. Effects of sex and a cafeteria diet. PLoS One 10:e0119572. https://doi.org/10.1371/journal.pone.0119572
Barbosa J, Faria J, Leal S, Afonso LP, Lobo J, Queirós O et al (2017) Acute administration of tramadol and tapentadol at effective analgesic and maximum tolerated doses causes hepato- and nephrotoxic effects in Wistar rats. Toxicology 389:118–129. https://doi.org/10.1016/j.tox.2017.07.001
Briffa JF, Grinfeld E, Jenkin KA, Mathai ML, Poronnik P, McAinch AJ et al (2015) Diet induced obesity in rats reduces NHE3 and Na(+) /K(+)-ATPase expression in the kidney. Clin Exp Pharmacol Physiol 42:1118–1126. https://doi.org/10.1111/1440-1681.12452
Byeon C-H, Kang K-Y, Kang S-H, Bae E-J (2015) Sarcopenia is associated with Framingham risk score in the Korean population: Korean national health and nutrition examination survey (KNHANES) 2010–2011. J Geriatr Cardiol 12:366–372. https://doi.org/10.11909/j.issn.1671-5411.2015.04.007
Catrysse L, van Loo G (2017) Inflammation and the metabolic syndrome: the tissue-specific functions of NF-κB. Trends Cell Biol 27:417–429. https://doi.org/10.1016/j.tcb.2017.01.006
Cha MC, Chou CJ, Boozer CN (2000) High-fat diet feeding reduces the diurnal variation of plasma leptin concentration in rats. Metabolism 49:503–507. https://doi.org/10.1016/S0026-0495(00)80016-5
Christ A, Lauterbach M, Latz E (2019) Western diet and the immune system: an inflammatory connection. Immunity 51:794–811. https://doi.org/10.1016/j.immuni.2019.09.020
Cotter TG, Rinella M (2020) Nonalcoholic fatty liver disease 2020: the state of the disease. Gastroenterology 158:1851–1864. https://doi.org/10.1053/j.gastro.2020.01.052
Cox CL, Stanhope KL, Schwarz JM, Graham JL, Hatcher B, Griffen SC et al (2012) Consumption of fructose-sweetened beverages for 10 weeks reduces net fat oxidation and energy expenditure in overweight/obese men and women. Eur J Clin Nutr 66:201–208. https://doi.org/10.1038/ejcn.2011.159
Da Silva-Santi LG, Antunes MM, Caparroz-Assef SM, Carbonera F, Masi LN, Curi R et al (2016) Liver fatty acid composition and inflammation in mice fed with high-carbohydrate diet or high-fat diet. Nutrients 8:682. https://doi.org/10.3390/nu8110682
De Marco P, Henriques AC, Azevedo R, Sá SI, Cardoso A, Fonseca B et al (2021) Gut microbiome composition and metabolic status are differently affected by early exposure to unhealthy diets in a rat model. Nutrients 13:3236. https://doi.org/10.3390/nu13093236
Digirolamo M, Newby FD, Lovejoy J (1992) Lactate production in adipose tissue; a regulated function with extra-adipose implications. FASEB J 6:2405–2412. https://doi.org/10.1096/fasebj.6.7.1563593
Duque-Guimarães DE, Ozanne SE (2013) Nutritional programming of insulin resistance: causes and consequences. Trends Endocrinol Metab 24:525–535. https://doi.org/10.1016/j.tem.2013.05.006
El-Haschimi K, Pierroz DD, Hileman SM, Bjørbæk C, Flier JS (2000) Two defects contribute to hypothalamic leptin resistance in mice with diet-induced obesity. J Clin Invest 105:1827–1832. https://doi.org/10.1172/JCI9842
Fernández-Sánchez A, Madrigal-Santillán E, Bautista M, Esquivel-Soto J, Morales-González Á, Esquivel-Chirino C et al (2011) Inflammation, oxidative stress, and obesity. Int J Mol Sci 12:3117–3132. https://doi.org/10.3390/ijms12053117
Francés DE, Motiño O, Agrá N, González-Rodríguez Á, Fernández-Álvarez A, Cucarella C et al (2015) Hepatic cyclooxygenase-2 expression protects against diet-induced steatosis, obesity, and insulin resistance. Diabetes 64:1522–1531. https://doi.org/10.2337/db14-0979
Francisqueti FV, Nascimento AF, Minatel IO, Dias MC, de Luvizotto RAM, Berchieri-Ronchi C et al (2017) Metabolic syndrome and inflammation in adipose tissue occur at different times in animals submitted to a high-sugar/fat diet. J Nutr Sci. https://doi.org/10.1017/jns.2017.42
Gluchowski NL, Becuwe M, Walther TC, Farese RV (2017) Lipid droplets and liver disease: from basic biology to clinical implications. Nat Rev Gastroenterol Hepatol 14:343–355. https://doi.org/10.1038/nrgastro.2017.32
Godoy-Matos AF, Silva Júnior WS, Valerio CM (2020) NAFLD as a continuum: from obesity to metabolic syndrome and diabetes. Diabetol Metab Syndr 12:60. https://doi.org/10.1186/s13098-020-00570-y
Gregor MF, Hotamisligil GS (2011) Inflammatory mechanisms in obesity. Annu Rev Immunol 29:415–445. https://doi.org/10.1146/annurev-immunol-031210-101322
Hardwick JP, Eckman K, Lee YK, Abdelmegeed MA, Esterle A, Chilian WM et al (2013) Eicosanoids in metabolic syndrome. Adv Pharmacol 66:157–266. https://doi.org/10.1016/B978-0-12-404717-4.00005-6
Ignat S-R, Dinescu S, Hermenean A, Costache M (2020) Cellular interplay as a consequence of inflammatory signals leading to liver fibrosis development. Cells 9:461. https://doi.org/10.3390/cells9020461
Johnson AR, Wilkerson MD, Sampey BP, Troester MA, Hayes DN, Makowski L (2016) Cafeteria diet-induced obesity causes oxidative damage in white adipose. Biochem Biophys Res Commun 473:545–550. https://doi.org/10.1016/j.bbrc.2016.03.113
Kalinkovich A, Livshits G (2017) Sarcopenic obesity or obese sarcopenia: a cross talk between age-associated adipose tissue and skeletal muscle inflammation as a main mechanism of the pathogenesis. Ageing Res Rev 35:200–221. https://doi.org/10.1016/j.arr.2016.09.008
Kelly AS, Ryder JR, Marlatt KL, Rudser KD, Jenkins T, Inge TH (2016) Changes in inflammation, oxidative stress and adipokines following bariatric surgery among adolescents with severe obesity. Int J Obes 40:275–280. https://doi.org/10.1038/ijo.2015.174
Kennedy AJ, Ellacott KLJ, King VL, Hasty AH (2010) Mouse models of the metabolic syndrome. Dis Model Mech 3:156–166. https://doi.org/10.1242/dmm.003467
Kim JI, Huh JY, Sohn JH, Choe SS, Lee YS, Lim CY et al (2015) Lipid-overloaded enlarged adipocytes provoke insulin resistance independent of inflammation. Mol Cell Biol 35:1686–1699. https://doi.org/10.1128/MCB.01321-14
Kim D, Touros A, Kim WR (2018) Nonalcoholic fatty liver disease and metabolic syndrome. Clin Liver Dis 22:133–140. https://doi.org/10.1016/j.cld.2017.08.010
Könner AC, Brüning JC (2011) Toll-like receptors: linking inflammation to metabolism. Trends Endocrinol Metab 22:16–23. https://doi.org/10.1016/j.tem.2010.08.007
Könner AC, Brüning JC (2012) Selective insulin and leptin resistance in metabolic disorders. Cell Metab 16:144–152. https://doi.org/10.1016/j.cmet.2012.07.004
la Fleur SE, Luijendijk MCM, van der Zwaal EM, Brans MAD, Adan RAH (2014) The snacking rat as model of human obesity: effects of a free-choice high-fat high-sugar diet on meal patterns. Int J Obes 38:643–649. https://doi.org/10.1038/ijo.2013.159
Lazaris AC, Dicoglou C, Tseleni-Balafouta S, Paraskevakou H, Davaris PS (1999) In situ expression of E-selectin and intercellular adhesion molecule-1 in chronic inflammatory diseases of the gastrointestinal tract. APMIS 107:819–827. https://doi.org/10.1111/j.1699-0463.1999.tb01477.x
Leal S, Sá C, Gonçalves J, Fresco P, Diniz C (2008) Immunohistochemical characterization of adenosine receptors in rat aorta and tail arteries. Microsc Res Tech 71:703–709. https://doi.org/10.1002/jemt.20609
Lee EY, Yoon K-H (2018) Epidemic obesity in children and adolescents: risk factors and prevention. Front Med 12:658–666. https://doi.org/10.1007/s11684-018-0640-1
Lee A-H, Scapa EF, Cohen DE, Glimcher LH (2008) Regulation of hepatic lipogenesis by the transcription factor XBP1. Science 320:1492–1496. https://doi.org/10.1126/science.1158042
Lee M-J, Wu Y, Fried SK (2010) Adipose tissue remodeling in pathophysiology of obesity. Curr Opin Clin Nutr Metab Care 13:371–376. https://doi.org/10.1097/MCO.0b013e32833aabef
Liu T, Zhang L, Joo D, Sun S-C (2017) NF-κB signaling in inflammation. Signal Transduct Target Ther 2:1–9. https://doi.org/10.1038/sigtrans.2017.23
Liu H-M, Lee T-Y, Liao J-F (2018) GW4064 attenuates lipopolysaccharide-induced hepatic inflammation and apoptosis through inhibition of the Toll-like receptor 4-mediated p38 mitogen-activated protein kinase signaling pathway in mice. Int J Mol Med 41:1455–1462. https://doi.org/10.3892/ijmm.2018.3366
Maersk M, Belza A, Stødkilde-Jørgensen H, Ringgaard S, Chabanova E, Thomsen H et al (2012) Sucrose-sweetened beverages increase fat storage in the liver, muscle, and visceral fat depot: a 6-mo randomized intervention study. Am J Clin Nutr 95:283–289. https://doi.org/10.3945/ajcn.111.022533
Martín-Sanz P, Mayoral R, Casado M, Boscá L (2010) COX-2 in liver, from regeneration to hepatocarcinogenesis: what we have learned from animal models? World J Gastroenterol 16:1430–1435. https://doi.org/10.3748/wjg.v16.i12.1430
Mathis D (2013) Immunological goings-on in visceral adipose tissue. Cell Metab 17:851–859. https://doi.org/10.1016/j.cmet.2013.05.008
Moreno-Fernández S, Garcés-Rimón M, Vera G, Astier J, Landrier JF, Miguel M (2018) High fat/high glucose diet induces metabolic syndrome in an experimental rat model. Nutrients 10:1502. https://doi.org/10.3390/nu10101502
Motiño O, Francés DE, Casanova N, Fuertes-Agudo M, Cucarella C, Flores JM et al (2019) Protective role of hepatocyte cyclooxygenase-2 expression against liver ischemia-reperfusion injury in mice. Hepatology 70:650–665. https://doi.org/10.1002/hep.30241
Özcan U, Cao Q, Yilmaz E, Lee A-H, Iwakoshi NN, Özdelen E et al (2004) Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes. Science 306:457–461. https://doi.org/10.1126/science.1103160
Panasevich MR, Meers GM, Linden MA, Booth FW, Perfield JW, Fritsche KL et al (2018) High-fat, high-fructose, high-cholesterol feeding causes severe NASH and cecal microbiota dysbiosis in juvenile Ossabaw swine. Am J Physiol-Endocrinol Metab 314:E78–E92. https://doi.org/10.1152/ajpendo.00015.2017
Peng L, Wu S, Zhou N, Zhu S, Liu Q, Li X (2021) Clinical characteristics and risk factors of nonalcoholic fatty liver disease in children with obesity. BMC Pediatr 21:122. https://doi.org/10.1186/s12887-021-02595-2
Peverill W, Powell LW, Skoien R (2014) Evolving concepts in the pathogenesis of NASH: beyond steatosis and inflammation. Int J Mol Sci 15:8591–8638. https://doi.org/10.3390/ijms15058591
Pini RTB, Ferreira do Vales LDM, Braga Costa TM, Almeida SS (2017) Effects of cafeteria diet and high fat diet intake on anxiety, learning and memory in adult male rats. Nutr Neurosci 20:396–408. https://doi.org/10.1080/1028415X.2016.1149294
Porras D, Nistal E, Martínez-Flórez S, Pisonero-Vaquero S, Olcoz JL, Jover R et al (2017) Protective effect of quercetin on high-fat diet-induced non-alcoholic fatty liver disease in mice is mediated by modulating intestinal microbiota imbalance and related gut-liver axis activation. Free Radic Biol Med 102:188–202. https://doi.org/10.1016/j.freeradbiomed.2016.11.037
Rayner DV, Trayhurn P (2001) Regulation of leptin production: sympathetic nervous system interactions. J Mol Med Berl Ger 79:8–20. https://doi.org/10.1007/s001090100198
Ritze Y, Bárdos G, D’Haese JG, Ernst B, Thurnheer M, Schultes B et al (2014) Effect of high sugar intake on glucose transporter and weight regulating hormones in mice and humans. PLoS One 9:e101702. https://doi.org/10.1371/journal.pone.0101702
Rodrigues RM, He Y, Hwang S, Bertola A, Mackowiak B, Ait-Ahmed Y et al (2021) E-selectin-dependent inflammation and lipolysis in adipose tissue exacerbate steatosis-to-NASH progression via S100A8/9. Cell Mol Gastroenterol Hepatol. https://doi.org/10.1016/j.jcmgh.2021.08.002
Rogero MM, Calder PC (2018) Obesity, inflammation, toll-like receptor 4 and fatty acids. Nutrients 10:432. https://doi.org/10.3390/nu10040432
Sabater D, Arriarán S, del Mar Romero M, Agnelli S, Remesar X, Fernández-López JA et al (2014) Cultured 3T3L1 adipocytes dispose of excess medium glucose as lactate under abundant oxygen availability. Sci Rep 4:3663. https://doi.org/10.1038/srep03663
Schwartz GJ, Brion LP, Spitzer A (1987) The use of plasma creatinine concentration for estimating glomerular filtration rate in infants, children, and adolescents. Pediatr Clin North Am 34:571–590. https://doi.org/10.1016/S0031-3955(16)36251-4
Silva M, Videira PA, Sackstein R (2018) E-selectin ligands in the human mononuclear phagocyte system: implications for infection, inflammation, and immunotherapy. Front Immunol 8:1878. https://doi.org/10.3389/fimmu.2017.01878
Simons N, Bijnen M, Wouters KAM, Rensen SS, Beulens JWJ, van Greevenbroek MMJ et al (2020) The endothelial function biomarker soluble E-selectin is associated with nonalcoholic fatty liver disease. Liver Int 40:1079–1088. https://doi.org/10.1111/liv.14384
Sol VV, Fresno M (2005) Involvement of TNF and NF-κB in the transcriptional control of cyclooxygenase-2 expression by IFN-γ in macrophages. J Immunol 174:2825–2833. https://doi.org/10.4049/jimmunol.174.5.2825
van der Poorten D, Milner K-L, Hui J, Hodge A, Trenell MI, Kench JG et al (2008) Visceral fat: a key mediator of steatohepatitis in metabolic liver disease. Hepatology 48:449–457. https://doi.org/10.1002/hep.22350
Weihe P, Weihrauch-Blüher S (2019) Metabolic syndrome in children and adolescents: diagnostic criteria, therapeutic options and perspectives. Curr Obes Rep 8:472–479. https://doi.org/10.1007/s13679-019-00357-x
Weihrauch-Blüher S, Schwarz P, Klusmann J-H (2019) Childhood obesity: increased risk for cardiometabolic disease and cancer in adulthood. Metabolism 92:147–152. https://doi.org/10.1016/j.metabol.2018.12.001
Weiss IC, Pryce CR, Jongen-Rêlo AL, Nanz-Bahr NI, Feldon J (2004) Effect of social isolation on stress-related behavioural and neuroendocrine state in the rat. Behav Brain Res 152:279–295. https://doi.org/10.1016/j.bbr.2003.10.015
Xiao Y, Liu D, Cline MA, Gilbert ER (2020) Chronic stress, epigenetics, and adipose tissue metabolism in the obese state. Nutr Metab 17:88. https://doi.org/10.1186/s12986-020-00513-4
Yang X, Schnackenberg LK, Shi Q, Salminen WF (2014) Chapter 13—hepatic toxicity biomarkers. In: Gupta RC (ed) Biomarkers in toxicology. Academic, Boston, pp 241–259. https://doi.org/10.1016/B978-0-12-404630-6.00013-0
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This work was supported by ERDF through the operation POCI-01-0145-FEDER-007746 funded by the Programa Operacional Competitividade e Internacionalização—COMPETE2020 and by National Funds through FCT—Fundação para a Ciência e a Tecnologia, I.P., within CINTESIS, R&D Unit (reference UIDB/4255/2020). This work is financed by national funds from FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. Sandra Leal was supported by a research grant from CESPU-IINFACTS [grant number FoodMicrobiome_CESPU_2017].
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SN contributed with laboratory work, data analysis and interpretation, preparation of the figures and the manuscript. BMF contributed with data analysis and interpretation and revising the manuscript. RS contributed with data analysis and interpretation and revising the manuscript. SL contributed to the conception and preparation of the study, data analysis and interpretation, and revising the manuscript. FG and LCM contributed with laboratory work and data analysis. AC contributed to the conception of the experimental model and revising the manuscript. SIS contributed with the conception and preparation of the study, laboratory work, data analysis and interpretation and revising the manuscript. All authors approved the final version of the manuscript.
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Nogueira, S., Garcez, F., Sá, S. et al. Early unhealthy eating habits underlie morpho-functional changes in the liver and adipose tissue in male rats. Histochem Cell Biol 157, 657–669 (2022). https://doi.org/10.1007/s00418-022-02092-2
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DOI: https://doi.org/10.1007/s00418-022-02092-2