{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:17:49Z","timestamp":1788499069970,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T00:00:00Z","timestamp":1787961600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>The nephrotoxicity and ototoxicity of Gentamicin have been extensively investigated; however, whether they induce hematotoxicity remains unclear. This study aimed to explore the binding of Gentamicin to adult hemoglobin (HbA) and its toxicological implications. The effect of Gentamicin on HbA oxidation was assessed by quantifying methemoglobin (MetHb) formation via a four-wavelength spectrophotometric method, and scavenger rescue assays were employed to elucidate the oxidative mechanism. Alterations in the oxygen-carrying capacity of both HbA and RBCs were evaluated through the acquisition of oxygen equilibrium curves and oxygen dissociation assays. The binding affinity between Gentamicin and HbA was determined by surface plasmon resonance (SPR). Furthermore, the influence of Gentamicin on the secondary and tertiary structures of HbA was examined by microfluidic modulation spectroscopy (MMS) and UV\u2013visible absorption spectroscopy, respectively. Molecular docking was employed to predict the binding sites of Gentamicin on HbA. Molecular dynamics simulations verified the stability of the binding. Gentamicin promoted HbA autoxidation, elevating MetHb levels, and reduced the oxygen affinity of HbA. Gentamicin-promoted HbA autoxidation is primarily mediated by both direct heme-pocket perturbation and H2O2\/iron-dependent amplification. SPR confirmed concentration-dependent specific binding between Gentamicin and HbA. MMS indicated no alteration in HbA secondary structure; however, UV\u2013visible spectroscopy revealed that Gentamicin attenuated the Soret band, converted the oxyhemoglobin double-peak to a singlet, and generated a new band at 630 nm. Molecular docking predicted that Gentamicin binds \u03b2-chain residues via hydrogen, carbon\u2013hydrogen, and hydrophobic interactions. This study reveals that Gentamicin exerts hematological effects in vitro. By binding to specific sites on HbA, Gentamicin affects the tertiary structure of HbA, promotes heme oxidation, ultimately increases MetHb content (oxidative damage), and decreases its oxygen-carrying capacity (functional impairment).<\/jats:p>","DOI":"10.3390\/ijms27177760","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:48:36Z","timestamp":1788256116000},"page":"7760","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Gentamicin Targeting Human Hemoglobin Induces Methemoglobin Formation and Decreases Oxygen Affinity: A Molecular Mechanism of Hematologic Toxicity"],"prefix":"10.3390","volume":"27","author":[{"given":"Peilin","family":"Shu","sequence":"first","affiliation":[{"name":"Air Force Medical Center, Air Force Medical University, Beijing 100142, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6298-7478","authenticated-orcid":false,"given":"Pengfei","family":"Wang","sequence":"additional","affiliation":[{"name":"Air Force Medical Center, Air Force Medical University, Beijing 100142, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wencong","family":"Li","sequence":"additional","affiliation":[{"name":"Air Force Medical Center, Air Force Medical University, Beijing 100142, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baichuan","family":"Gu","sequence":"additional","affiliation":[{"name":"School of Integrated Circuits, Beijing University of Technology, Beijing 100124, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanjing","family":"Zheng","sequence":"additional","affiliation":[{"name":"Air Force Medical Center, Air Force Medical University, Beijing 100142, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"Academy of Military Medical Sciences, No. 27 Taiping Road, Haidian, Beijing 100850, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minghao","family":"Yang","sequence":"additional","affiliation":[{"name":"Air Force Medical Center, Air Force Medical University, Beijing 100142, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9790-1256","authenticated-orcid":false,"given":"Lian","family":"Zhao","sequence":"additional","affiliation":[{"name":"Academy of Military Medical Sciences, No. 27 Taiping Road, Haidian, Beijing 100850, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"133","DOI":"10.18773\/austprescr.2025.038","article-title":"Aminoglycosides: An update on indications, dosing and monitoring","volume":"48","author":"Moore","year":"2025","journal-title":"Aust. Prescr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1016\/j.otc.2021.08.007","article-title":"Mechanisms of Ototoxicity and Otoprotection","volume":"54","author":"Steyger","year":"2021","journal-title":"Otolaryngol. Clin. N. Am."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1016\/S0272-6386(97)90212-2","article-title":"Oxidant mechanisms in toxic acute renal failure","volume":"29","author":"Baliga","year":"1997","journal-title":"Am. J. Kidney Dis."},{"key":"ref_4","unstructured":"AURA (2023). Fifth Australian Report on Antimicrobial Use and Resistance in Human Health."},{"key":"ref_5","first-page":"1","article-title":"Australian Group on Antimicrobial Resistance (AGAR) Australian Gram-negative Surveillance Outcome Program (GnSOP) Bloodstream Infection Annual Report 2023","volume":"49","author":"Bell","year":"2025","journal-title":"Commun. Dis. Intell."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1016\/j.freeradbiomed.2011.07.015","article-title":"Role of oxidative stress in lysosomal membrane permeabilization and apoptosis induced by gentamicin, an aminoglycoside antibiotic","volume":"51","author":"Denamur","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1111\/bcp.13439","article-title":"Adverse effects of a single dose of gentamicin in adults: A systematic review","volume":"84","author":"Hayward","year":"2018","journal-title":"Br. J. Clin. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1731","DOI":"10.1093\/cid\/cix186","article-title":"Short-Course Adjunctive Gentamicin as Empirical Therapy in Patients With Severe Sepsis and Septic Shock: A Prospective Observational Cohort Study","volume":"64","author":"Ong","year":"2017","journal-title":"Clin. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1016\/j.jcf.2017.01.006","article-title":"The cumulative effects of intravenous antibiotic treatments on hearing in patients with cystic fibrosis","volume":"16","author":"Garinis","year":"2017","journal-title":"J. Cyst. Fibros"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.ijporl.2017.03.025","article-title":"Effect of gentamicin and levels of ambient sound on hearing screening outcomes in the neonatal intensive care unit: A pilot study","volume":"97","author":"Garinis","year":"2017","journal-title":"Int. J. Pediatr. Otorhinolaryngol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.jcf.2020.08.002","article-title":"The implementation of an aminoglycoside induced ototoxicity algorithm for people with cystic fibrosis","volume":"20","author":"Elson","year":"2021","journal-title":"J. Cyst. Fibros"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.etap.2016.11.004","article-title":"Protective effect of quercetin in gentamicin-induced oxidative stress in vitro and in vivo in blood cells. Effect on gentamicin antimicrobial activity","volume":"48","author":"Bustos","year":"2016","journal-title":"Environ. Toxicol. Pharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.taap.2014.02.022","article-title":"Naringin ameliorates gentamicin-induced nephrotoxicity and associated mitochondrial dysfunction, apoptosis and inflammation in rats: Possible mechanism of nephroprotection","volume":"277","author":"Sahu","year":"2014","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1016\/j.ijporl.2011.09.027","article-title":"Calcium imaging in gentamicin ototoxicity: Increased intracellular calcium relates to oxidative stress and late apoptosis","volume":"75","author":"Chang","year":"2011","journal-title":"Int. J. Pediatr. Otorhinolaryngol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"106393","DOI":"10.1016\/j.biocel.2023.106393","article-title":"The endocytosis receptor megalin: From bench to bedside","volume":"157","author":"Goto","year":"2023","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s10162-009-0160-4","article-title":"Trafficking of systemic fluorescent gentamicin into the cochlea and hair cells","volume":"10","author":"Wang","year":"2009","journal-title":"J. Assoc. Res. Otolaryngol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"101037","DOI":"10.1016\/j.mam.2021.101037","article-title":"Hemoglobin allostery and pharmacology","volume":"84","author":"Bellelli","year":"2022","journal-title":"Mol. Asp. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/0076-6879(90)86118-F","article-title":"Oxidative reactions of hemoglobin","volume":"186","author":"Winterbourn","year":"1990","journal-title":"Methods Enzymol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/j.revmed.2024.05.001","article-title":"M\u00e9th\u00e9moglobin\u00e9mie","volume":"45","author":"Batton","year":"2024","journal-title":"La Rev. M\u00e9d. Interne"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7442","DOI":"10.1080\/07391102.2023.2245043","article-title":"Cefmetazole sodium as an allosteric effector that regulates the oxygen supply efficiency of adult hemoglobin","volume":"42","author":"Shu","year":"2024","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"101022","DOI":"10.1016\/j.mam.2021.101022","article-title":"Role of hemoglobin structural-functional relationships in oxygen transport","volume":"84","author":"Ciaccio","year":"2022","journal-title":"Mol. Asp. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1016\/j.jhazmat.2010.04.110","article-title":"Toxic interaction mechanism between oxytetracycline and bovine hemoglobin","volume":"180","author":"Chi","year":"2010","journal-title":"J. Hazard. Mater."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"401","DOI":"10.6026\/97320630002401","article-title":"An in silico approach to map the binding site of doxorubicin on hemoglobin","volume":"2","author":"Khan","year":"2008","journal-title":"Bioinformation"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"735","DOI":"10.3109\/03630268008997741","article-title":"The effects of adriamycin (doxorubicin HCl) on human red blood cells","volume":"4","author":"Shinohara","year":"1980","journal-title":"Hemoglobin"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7786","DOI":"10.1080\/07391102.2021.1902398","article-title":"Spectroscopic investigation on molecular aspects and structural and functional effects of tetraethyl pyrophosphate organophosphorus insecticide interaction with adult human hemoglobin","volume":"40","author":"Doroudian","year":"2022","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Bansal, S., and SenGupta, U. (2017). A Study of the Interaction of Bovine Hemoglobin with Synthetic Dyes Using Spectroscopic Techniques and Molecular Docking. Front. Chem., 4.","DOI":"10.3389\/fchem.2016.00050"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"119","DOI":"10.2174\/138161207779313731","article-title":"Aminoglycoside-induced ototoxicity","volume":"13","author":"Selimoglu","year":"2007","journal-title":"Curr. Pharm. Des."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4349","DOI":"10.1016\/0006-2952(89)90641-2","article-title":"Reduced glutathione in combination with superoxide dismutase as an important biological antioxidant defence mechanism","volume":"38","author":"Munday","year":"1989","journal-title":"Biochem. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"153832","DOI":"10.1016\/j.tox.2024.153832","article-title":"The impact of cannabinoids on methemoglobin formation and hemoglobin oxygen affinity: An ex-vivo study","volume":"505","author":"Frisch","year":"2024","journal-title":"Toxicology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0301-4622(90)88017-M","article-title":"Oxygen binding to partially oxidized hemoglobin: Analysis in terms of an allosteric model","volume":"37","author":"Cordone","year":"1990","journal-title":"Biophys. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1016\/0731-7085(95)01509-J","article-title":"A 19F NMR study of lomefloxacin in human erythrocytes and its interaction with hemoglobin","volume":"13","author":"Knaub","year":"1995","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4441","DOI":"10.1016\/S0021-9258(18)99558-X","article-title":"The heme chromophore in the ultraviolet","volume":"242","author":"Urry","year":"1967","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2174","DOI":"10.1021\/bi00707a027","article-title":"Influence of globin structure on the state of the heme. II. Allosteric transitions in methemoglobin","volume":"13","author":"Perutz","year":"1974","journal-title":"Biochemistry"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1016\/0005-2795(76)90234-8","article-title":"Dependence of magneto-optical rotatory dispersion and magnetic circular dichroism of deoxy- and methemoglobin on their quaternary structure","volume":"434","author":"Sharonov","year":"1976","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_35","first-page":"C2-631","article-title":"Spin Resolved Multiple Scattering Study on Iron Spin Transition in Metmyoglobin","volume":"7","author":"Bianconi","year":"1997","journal-title":"J. Phys. IV Fr."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3057","DOI":"10.1016\/S0021-9258(18)94029-9","article-title":"Discrimination between Oxygen and Carbon Monoxide and Inhibition of Autooxidation by Myoglobin: Site-directed mutagenesis of the distal histidine","volume":"264","author":"Springer","year":"1989","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"25512","DOI":"10.1074\/jbc.M113.474841","article-title":"Autoxidation and oxygen binding properties of recombinant hemoglobins with substitutions at the \u03b1Val-62 or \u03b2Val-67 position of the distal heme pocket","volume":"288","author":"Tam","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1080\/21691401.2020.1770272","article-title":"The P50 value detected by the oxygenation-dissociation analyser and blood gas analyser","volume":"48","author":"Chu","year":"2020","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/0003-2697(73)90309-6","article-title":"Equations for the spectrophotometric analysis of hemoglobin mixtures","volume":"55","author":"Benesch","year":"1973","journal-title":"Anal. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.2174\/15665240113139990054","article-title":"Suicidal inactivation of methemoglobin by generation of thiyl radical: Insight into NAC mediated protection in RBC","volume":"13","author":"Balaji","year":"2013","journal-title":"Curr. Mol. Med."},{"key":"ref_41","first-page":"609","article-title":"Cross-linked hemoglobin-superoxide dismutase-catalase scavenges oxygen-derived free radicals and prevents methemoglobin formation and iron release","volume":"21","author":"Chang","year":"1993","journal-title":"Biomater. Artif. Cells Immobil. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1007\/s12975-012-0195-4","article-title":"Effect of iron chelators on methemoglobin and thrombin preconditioning","volume":"3","author":"Sinanan","year":"2012","journal-title":"Transl. Stroke Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"215","DOI":"10.3109\/03009738809178547","article-title":"Inhibition of heme-promoted enzymatic lipid peroxidation by desferrioxamine and EDTA","volume":"93","author":"Carlin","year":"1988","journal-title":"Ups. J. Med. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1599","DOI":"10.2147\/DDDT.S157570","article-title":"Development and validation of an oxygen dissociation assay, a screening platform for discovering, and characterizing hemoglobin-oxygen affinity modifiers","volume":"12","author":"Patel","year":"2018","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"113762","DOI":"10.1016\/j.ejmech.2021.113762","article-title":"Discovery of novel thiophene derivatives as potent neuraminidase inhibitors","volume":"225","author":"Zhong","year":"2021","journal-title":"Eur. J. Med. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"118530","DOI":"10.1016\/j.saa.2020.118530","article-title":"Probing the structure-function relationship of hemoglobin in living human red blood cells","volume":"239","author":"Dybas","year":"2020","journal-title":"Spectrochim. Acta A Mol. Biomol. Spectrosc."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"117151","DOI":"10.1016\/j.jep.2023.117151","article-title":"Neuroprotective and anti-amnesic effects of Laurus Nobilis essential oil against scopolamine-induced memory deficits in mice brain","volume":"319","author":"Smach","year":"2024","journal-title":"J. Ethnopharmacol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7760\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T04:34:49Z","timestamp":1788496489000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7760"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,29]]},"references-count":47,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27177760"],"URL":"https:\/\/doi.org\/10.3390\/ijms27177760","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,29]]}}}