{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T10:32:34Z","timestamp":1788172354937,"version":"build-2803163510"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,10]],"date-time":"2022-02-10T00:00:00Z","timestamp":1644451200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Association de D\u00e9fense des Insuffisants R\u00e9naux\u201d (ADIR, Brussels, Belgium) and  European Regional Development Fund and the Walloon Region, Belgium.","award":["none"],"award-info":[{"award-number":["none"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Aristolochic acids (AAs) are powerful nephrotoxins that cause severe tubulointerstitial fibrosis. The biopsy-proven peritubular capillary rarefaction may worsen the progression of renal lesions via tissue hypoxia. As we previously observed the overproduction of reactive oxygen species (ROS) by cultured endothelial cells exposed to AA, we here investigated in vitro AA-induced metabolic changes by 1H-NMR spectroscopy on intracellular medium and cell extracts. We also tested the effects of nebivolol (NEB), a \u03b2-blocker agent exhibiting antioxidant properties. After 24 h of AA exposure, significantly reduced cell viability and intracellular ROS overproduction were observed in EAhy926 cells; both effects were counteracted by NEB pretreatment. After 48 h of exposure to AA, the most prominent metabolite changes were significant decreases in arginine, glutamate, glutamine and glutathione levels, along with a significant increase in the aspartate, glycerophosphocholine and UDP-N-acetylglucosamine contents. NEB pretreatment slightly inhibited the changes in glutathione and glycerophosphocholine. In the supernatants from exposed cells, a decrease in lactate and glutamate levels, together with an increase in glucose concentration, was found. The AA-induced reduction in glutamate was significantly inhibited by NEB. These findings confirm the involvement of oxidative stress in AA toxicity for endothelial cells and the potential benefit of NEB in preventing endothelial injury.<\/jats:p>","DOI":"10.3390\/toxins14020132","type":"journal-article","created":{"date-parts":[[2022,2,11]],"date-time":"2022-02-11T02:40:17Z","timestamp":1644547217000},"page":"132","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Protective Effect of Nebivolol against Oxidative Stress Induced by Aristolochic Acids in Endothelial Cells"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8600-1818","authenticated-orcid":false,"given":"Marie-H\u00e9l\u00e8ne","family":"Antoine","sequence":"first","affiliation":[{"name":"Laboratory of Experimental Nephrology, Faculty of Medicine, Universit\u00e9 Libre de Bruxelles, Erasme Campus, 808 Route de Lennik, B-1070 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C\u00e9cile","family":"Husson","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Nephrology, Faculty of Medicine, Universit\u00e9 Libre de Bruxelles, Erasme Campus, 808 Route de Lennik, B-1070 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiana","family":"Yankep","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Nephrology, Faculty of Medicine, Universit\u00e9 Libre de Bruxelles, Erasme Campus, 808 Route de Lennik, B-1070 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Souhaila","family":"Mahria","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Nephrology, Faculty of Medicine, Universit\u00e9 Libre de Bruxelles, Erasme Campus, 808 Route de Lennik, B-1070 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9706-2045","authenticated-orcid":false,"given":"Vanessa","family":"Tagliatti","sequence":"additional","affiliation":[{"name":"Laboratory of Human Toxicology, University of Mons (UMONS), 6 Avenue du Champ de Mars, B-7000 Mons, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4440-2795","authenticated-orcid":false,"given":"Jean-Marie","family":"Colet","sequence":"additional","affiliation":[{"name":"Laboratory of Human Toxicology, University of Mons (UMONS), 6 Avenue du Champ de Mars, B-7000 Mons, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00eblle","family":"Nortier","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Nephrology, Faculty of Medicine, Universit\u00e9 Libre de Bruxelles, Erasme Campus, 808 Route de Lennik, B-1070 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/0140-6736(93)92984-2","article-title":"Rapidly progressive interstitial renal fibrosis in young women: Association with slimming regimen including Chinese herbs","volume":"13","author":"Vanherweghem","year":"1993","journal-title":"Lancet"},{"key":"ref_2","first-page":"340","article-title":"Nephrotoxicity of Natural Products: Aristolochic Acid and Fungal Toxins","volume":"Volume 14","author":"McQueen","year":"2018","journal-title":"Comprehensive Toxicology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1681\/ASN.V132431","article-title":"Aristolochic acids induce chronic renal failure with interstitial fibrosis in salt-depleted rats","volume":"13","author":"Debelle","year":"2002","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1159\/000094778","article-title":"Role of peritubular capillary loss and hypoxia in progressive tubulointerstitial fibrosis in a rat model of aristolochic acid nephropathy","volume":"26","author":"Sun","year":"2006","journal-title":"Am. J. Nephrol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/S0272-6386(12)80178-8","article-title":"Pathologic aspects of a newly described nephropathy related to the prolonged use of Chinese herbs","volume":"24","author":"Depierreux","year":"1994","journal-title":"Am. J. Kidney Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"F1383","DOI":"10.1152\/ajprenal.00366.2019","article-title":"Capillary rarefaction is more closely associated with CKD progression after cisplatin, rhabdomyolysis, and ischemia-reperfusion-induced AKI than renal fibrosis","volume":"317","author":"Menshikh","year":"2019","journal-title":"Am. J. Physiol. Renal Physiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1038\/ki.2008.350","article-title":"Chronic hypoxia as a mechanism of progression of chronic kidney diseases: From hypothesis to novel therapeutics","volume":"74","author":"Fine","year":"2008","journal-title":"Kidney Int."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1097\/00041552-200401000-00001","article-title":"Rarefaction of peritubular capillaries following ischemic acute renal failure: A potential factor predisposing to progressive nephropathy","volume":"13","author":"Basile","year":"2004","journal-title":"Curr. Opin. Nephrol. Hypertens."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1038\/aps.2011.74","article-title":"Aristolochic acid induces apoptosis of human umbilical vein endothelial cells in vitro by suppressing PI3K\/Akt signaling pathway","volume":"32","author":"Shi","year":"2011","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"104811","DOI":"10.1016\/j.tiv.2020.104811","article-title":"Characterization of cytotoxic effects of aristolochic acids on the vascular endothelium","volume":"65","author":"Youl","year":"2020","journal-title":"Toxicol. In Vitro"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1159\/000322334","article-title":"Differential effects of NO inhibition in renal epithelial and endothelial cells in mono-culture vs. co-culture conditions","volume":"26","author":"Bertocchi","year":"2010","journal-title":"Cell Physiol. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Dubois-Deruy, E., Peugnet, V., Turkieh, A., and Pinet, F. (2020). Oxidative Stress in Cardiovascular Diseases. Antioxidants, 9.","DOI":"10.3390\/antiox9090864"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"108917","DOI":"10.1016\/j.biopha.2019.108917","article-title":"Protective role of relaxin in a mouse model of aristolochic acid nephropathy","volume":"115","author":"Yang","year":"2019","journal-title":"Biomed. Pharmacother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1113\/EP085333","article-title":"Protective effect of nitric oxide in aristolochic acid-induced toxic acute kidney injury: An old friend with new assets","volume":"101","author":"Jadot","year":"2016","journal-title":"Exp. Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1038\/jhh.2017.8","article-title":"Protective effects of nebivolol from oxidative stress to prevent hypertension-related target organ damage","volume":"31","author":"Coats","year":"2017","journal-title":"J. Hum. Hypertens."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1016\/j.phrs.2006.12.008","article-title":"Effect of DL-nebivolol, its enantiomers and metabolites on the intracellular production of superoxide and nitric oxide in human endothelial cells","volume":"55","author":"Evangelista","year":"2007","journal-title":"Pharmacol. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/j.coph.2012.12.002","article-title":"Beta blockers, nitric oxide, and cardiovascular disease","volume":"13","author":"Vanhoutte","year":"2013","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1111\/j.1440-1681.2006.04424.x","article-title":"Mechanisms underlying nebivolol-induced endothelial nitric oxide synthase activation in human umbilical vein endothelial cells","volume":"33","author":"Ladage","year":"2006","journal-title":"Clin. Exp. Pharmacol. Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.copbio.2016.08.001","article-title":"The future of NMR-based metabolomics","volume":"43","author":"Markley","year":"2017","journal-title":"Curr. Opin. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.ddtec.2015.01.002","article-title":"Metabonomics in the preclinical and environmental toxicity field","volume":"13","author":"Colet","year":"2015","journal-title":"Drug Discov. Today Technol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.fct.2017.07.015","article-title":"Interest of metabonomic approach in environmental nephrotoxicants: Application to aristolochic acid exposure","volume":"108","author":"Duquesne","year":"2017","journal-title":"Food Chem. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"815","DOI":"10.3390\/toxins3070815","article-title":"Comparative (1)H NMR metabolomic urinalysis of people diagnosed with Balkan endemic nephropathy, and healthy subjects, in Romania and Bulgaria: A pilot study","volume":"3","author":"Mantle","year":"2011","journal-title":"Toxins"},{"key":"ref_23","first-page":"208","article-title":"Phenotypic differences in nebivolol metabolism and bioavailability in healthy volunteers","volume":"88","author":"Briciu","year":"2015","journal-title":"Clujul Med."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Pozdzik, A.A., Giordano, L., Li, G., Antoine, M.H., Quellard, N., Godet, J., De Prez, E., Husson, C., Decl\u00e8ves, A.E., and Arlt, V.M. (2016). Blocking TGF-\u03b2 Signaling Pathway Preserves Mitochondrial Proteostasis and Reduces Early Activation of PDGFR\u03b2+ Pericytes in Aristolochic Acid Induced Acute Kidney Injury in Wistar Male Rats. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0157288"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.tiv.2012.03.005","article-title":"Endoplasmic reticulum stress mediates aristolochic acid I-induced apoptosis in human renal proximal tubular epithelial cells","volume":"26","author":"Zhu","year":"2012","journal-title":"Toxicol. In Vitro"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1177\/0960327119897099","article-title":"Mitochondrial dysfunction is involved in aristolochic acid I-induced apoptosis in renal proximal tubular epithelial cells","volume":"39","author":"Liu","year":"2020","journal-title":"Hum. Exp. Toxicol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1097\/01.fjc.0000238593.67191.e2","article-title":"Effect of nebivolol on endothelial nitric oxide and peroxynitrite release in hypertensive animals: Role of antioxidant activity","volume":"48","author":"Mason","year":"2006","journal-title":"J. Cardiovasc. Pharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1097\/00005344-200401000-00022","article-title":"Antioxidant activity of nebivolol in the rat aorta","volume":"43","author":"Mathy","year":"2004","journal-title":"J. Cardiovasc. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1111\/1440-1681.13001","article-title":"Nebivolol ameliorated kidney damage in Zucker diabetic fatty rats by regulation of oxidative stress\/NO pathway: Comparison with captopril","volume":"45","author":"Wang","year":"2018","journal-title":"Clin. Exp. Pharmacol. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"79","DOI":"10.3389\/fphar.2020.00079","article-title":"New Insights About Doxorubicin-Induced Toxicity to Cardiomyoblast-Derived H9C2 Cells and Dexrazoxane Cytoprotective Effect: Contribution of In Vitro (1)H-NMR Metabonomics","volume":"11","author":"Dallons","year":"2020","journal-title":"Front Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1152\/physrev.00001.2017","article-title":"Endothelial Cell Metabolism","volume":"98","author":"Eelen","year":"2018","journal-title":"Physiol. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1007\/s00204-014-1200-3","article-title":"Integrated proteomic and metabolomic analysis to assess the effects of pure and benzo[a]pyrene-loaded carbon black particles on energy metabolism and motility in the human endothelial cell line EA.hy926","volume":"88","author":"Pink","year":"2014","journal-title":"Arch. Toxicol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/j.cell.2013.06.037","article-title":"Role of PFKFB3-driven glycolysis in vessel sprouting","volume":"154","author":"Georgiadou","year":"2013","journal-title":"Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s00424-012-1085-z","article-title":"Endothelial mitochondria\u2014Less respiration, more integration","volume":"464","author":"Groschner","year":"2012","journal-title":"Pflugers Arch."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1021\/acs.chemrestox.9b00488","article-title":"Identifying Cysteine, N-Acetylcysteine, and Glutathione Conjugates as Novel Metabolites of Aristolochic Acid I: Emergence of a New Detoxification Pathway","volume":"33","author":"Zhang","year":"2020","journal-title":"Chem. Res. Toxicol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.niox.2020.12.003","article-title":"Arginine recycling in endothelial cells is regulated BY HSP90 and the ubiquitin proteasome system","volume":"1","author":"Wu","year":"2021","journal-title":"Nitric Oxide"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"8612","DOI":"10.1073\/pnas.87.21.8612","article-title":"The metabolism of L-arginine and its significance for the biosynthesis of endothelium-derived relaxing factor: Cultured endothelial cells recycle L-citrulline to L-arginine","volume":"87","author":"Hecker","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1002\/path.2769","article-title":"Asymmetric dimethylarginine (ADMA) induces chronic kidney disease through a mechanism involving collagen and TGF-\u03b21 synthesis","volume":"223","author":"Mihout","year":"2011","journal-title":"J. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.taap.2019.04.014","article-title":"Untargeted LC-MS-based metabonomics revealed that aristolochic acid I induces testicular toxicity by inhibiting amino acids metabolism, glucose metabolism, beta-oxidation of fatty acids and the TCA cycle in male mice","volume":"373","author":"Cui","year":"2019","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1007\/s11064-016-1966-1","article-title":"Multifactorial Effects on Different Types of Brain Cells Contribute to Ammonia Toxicity","volume":"42","author":"Hertz","year":"2017","journal-title":"Neurochem. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1093\/toxsci\/kfq206","article-title":"Glycosylation protects proteins against free radicals generated from toxic xenobiotics","volume":"117","author":"Sulc","year":"2010","journal-title":"Toxicol. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1016\/j.apsb.2019.12.004","article-title":"In situ metabolomics in nephrotoxicity of aristolochic acids based on air flow-assisted desorption electrospray ionization mass spectrometry imaging","volume":"10","author":"Wang","year":"2020","journal-title":"Acta Pharm. Sin. B."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"12936","DOI":"10.1038\/srep12936","article-title":"Metabolomics analysis reveals the association between lipid abnormalities and oxidative stress, inflammation, fibrosis, and Nrf2 dysfunction in aristolochic acid-induced nephropathy","volume":"7","author":"Zhao","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1007\/s007020070051","article-title":"Membrane breakdown in acute and chronic neurodegeneration: Focus on choline-containing phospholipids","volume":"107","author":"Klein","year":"2000","journal-title":"J. Neural. Transm."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Bonvallot, N., Tremblay-Franco, M., Chevrier, C., Canlet, C., Warembourg, C., Cravedi, J.P., and Cordier, S. (2013). Metabolomics tools for describing complex pesticide exposure in pregnant women in Brittany (France). PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0064433"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Zhang, Y.X., Yang, X., Zou, P., Du, P.F., Wang, J., Jin, F., Jin, M.J., and She, Y.X. (2016). Nonylphenol Toxicity Evaluation and Discovery of Biomarkers in Rat Urine by a Metabolomics Strategy through HPLC-QTOF-MS. Int. J. Environ. Res. Public Health, 13.","DOI":"10.3390\/ijerph13050501"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1038\/sj.ki.5002714","article-title":"Aristolochic acid induces proximal tubule apoptosis and epithelial to mesenchymal transformation","volume":"73","author":"Pozdzik","year":"2008","journal-title":"Kidney Int."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.febslet.2007.01.036","article-title":"Metabolic profiling using combined GC-MS and LC-MS provides a systems understanding of aristolochic acid-induced nephrotoxicity in rat","volume":"581","author":"Ni","year":"2007","journal-title":"FEBS Lett."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"C50","DOI":"10.1152\/ajpcell.1992.262.1.C50","article-title":"Cystine uptake and glutathione level in endothelial cells exposed to oxidative stress","volume":"262","author":"Miura","year":"1992","journal-title":"Am. J. Physiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1007\/s13238-020-00789-5","article-title":"Cystine transporter SLC7A11\/xCT in cancer: Ferroptosis, nutrient dependency, and cancer therapy","volume":"12","author":"Koppula","year":"2021","journal-title":"Protein Cell"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/14\/2\/132\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:17:27Z","timestamp":1760134647000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/14\/2\/132"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,10]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["toxins14020132"],"URL":"https:\/\/doi.org\/10.3390\/toxins14020132","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,10]]}}}