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TFA has historically been regarded as a compound of relatively low acute lethality, yet its adverse effects on sensitive targets at environmentally relevant levels are largely unknown. Due to extreme hydrophilicity and anionic form in the aqueous phase of blood, TFA manifests high accessibility to the innermost layer of vasculature. In this study, we investigated the vascular toxicity of TFA using human umbilical vein endothelial cell and zebrafish embryo models at concentrations of 0.8\u20132000 \u03bcg\/L. The results showed that TFA exposure reduced ATP content, lactate production, and glycolytic flux, concomitantly increasing pyruvate accumulation. The benchmark dose lower confidence limit (BMDL5) values ranged from 0.44 to 2.65 \u03bcg\/L, falling within the range of environmental levels. Enzyme kinetics and molecular docking indicated that TFA competitively inhibited lactate dehydrogenase (LDH) at the pyruvate-binding site. In addition, TFA inhibited endothelial migration, induced cellular F-actin disruption, and impaired vascular development in zebrafish larvae. The AI-aided vascular phenomics revealed the most prominent defect in the subintestinal venous plexus. A quantitative adverse outcome pathway was subsequently established, and key-event relationships were robustly fitted, with R2 values exceeding 0.72. This study unravels a metabolic perturbation pathway of TFA-induced vascular toxicity and reinforces the need to reassess the health risks of TFA.<\/jats:p>","DOI":"10.1021\/acs.est.6c06569","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T09:02:43Z","timestamp":1789635763000},"page":"27001-27011","source":"Crossref","is-referenced-by-count":0,"title":["Trifluoroacetic\nAcid Competitively Inhibits Lactate\nDehydrogenase to Disrupt Endothelial Glycolysis and Impair Vascular\nDevelopment"],"prefix":"10.1021","volume":"60","author":[{"given":"Xiaolian","family":"Cao","sequence":"first","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuting","family":"Huang","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenxin","family":"Li","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuxin","family":"Jiang","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junru","family":"Li","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingying","family":"Zhou","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaiqin","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangzhou Medical University , , ,","place":["Guangzhou, China, 511436"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Ma","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University , , ,","place":["Shanghai, China, 200240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Chu","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Panna","family":"Yang","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9680-0077","authenticated-orcid":true,"given":"Xiaoxing","family":"Kou","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510055"]},{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510055"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6496-8174","authenticated-orcid":true,"given":"Yitao","family":"Pan","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University , , ,","place":["Shanghai, China, 200240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8959-4260","authenticated-orcid":true,"given":"Shengtao","family":"Ma","sequence":"additional","affiliation":[{"name":"Guangzhou Medical University , , ,","place":["Guangzhou, China, 511436"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4908-5597","authenticated-orcid":true,"given":"Jiayin","family":"Dai","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University , , ,","place":["Shanghai, China, 200240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9000-7016","authenticated-orcid":true,"given":"Xifei","family":"Yang","sequence":"additional","affiliation":[{"name":"Shenzhen Center for Disease Control and Prevention , , ,","place":["Shenzhen, China, 518055"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3398-5657","authenticated-orcid":true,"given":"Yanhong","family":"Wei","sequence":"additional","affiliation":[{"name":"Sun Yat-sen University , , ,","place":["Guangzhou, China, 510080"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"issue":"45","key":"2026092909071781900_cit1","doi-asserted-by":"publisher","first-page":"19925","DOI":"10.1021\/acs.est.4c06189","article-title":"The Global Threat from the Irreversible\nAccumulation\nof Trifluoroacetic Acid (TFA)","volume":"58","author":"Arp","year":"2024","journal-title":"Environ. Sci.\nTechnol."},{"issue":"12","key":"2026092909071781900_cit2","doi-asserted-by":"publisher","DOI":"10.3390\/life15121825","article-title":"The Effects of Trifluoroacetic Acid\n(TFA) in Humans: A Rapid Review","volume":"15","author":"Wipplinger","year":"2025","journal-title":"Life"},{"issue":"D5","key":"2026092909071781900_cit3","doi-asserted-by":"publisher","first-page":"5747","DOI":"10.1029\/97JD02988","article-title":"Trifluoroacetic\nacid from degradation of HCFCs and\nHFCs: A three-dimensional modeling study","volume":"103","author":"Kotamarthi","year":"1998","journal-title":"J.\nGeophys. Res.: Atmos."},{"issue":"10","key":"2026092909071781900_cit4","doi-asserted-by":"publisher","first-page":"6380","DOI":"10.1021\/acs.est.1c07949","article-title":"Ultra-Short-Chain\nPFASs in the Sources of German Drinking\nWater: Prevalent, Overlooked, Difficult to Remove, and Unregulated","volume":"56","author":"Neuwald","year":"2022","journal-title":"Environ. Sci. Technol."},{"issue":"47","key":"2026092909071781900_cit5","doi-asserted-by":"publisher","first-page":"25138","DOI":"10.1021\/acs.est.5c09274","article-title":"Environmental Occurrence, Source\nIdentification, and Health Hazards of Ultrashort-Chain PFAS in the\nYangtze River Delta","volume":"59","author":"Miao","year":"2025","journal-title":"Environ. Sci. Technol."},{"key":"2026092909071781900_cit6","first-page":"6","volume-title":"Trinkwasserqualita\u0308t Bezu\u0308glich\nder PFAS-Ru\u0308cksta\u0308nde","author":"Brodmann","year":"2024"},{"key":"2026092909071781900_cit7","doi-asserted-by":"publisher","DOI":"10.1016\/j.envpol.2026.127829","article-title":"Open dumps as a critical\nsource of per- and polyfluoroalkyl substances in agricultural soils\nof Pakistan: evidence of trifluoroacetic acid dominance","volume":"396","author":"Amanat","year":"2026","journal-title":"Environ. Pollut."},{"issue":"7","key":"2026092909071781900_cit8","doi-asserted-by":"publisher","first-page":"3675","DOI":"10.1021\/es4050264","article-title":"Airborne trifluoroacetic\nacid and its fraction from the degradation of HFC-134a in Beijing,\nChina","volume":"48","author":"Wu","year":"2014","journal-title":"Environ. Sci. Technol."},{"issue":"5","key":"2026092909071781900_cit9","doi-asserted-by":"publisher","first-page":"400","DOI":"10.1021\/acs.estlett.2c00164","article-title":"Levels and\nTemporal Trends of Trifluoroacetate (TFA)\nin Archived Plants: Evidence for Increasing Emissions of Gaseous TFA\nPrecursors over the Last Decades","volume":"9","author":"Freeling","year":"2022","journal-title":"Environ. Sci.\nTechnol. Lett."},{"key":"2026092909071781900_cit10","doi-asserted-by":"publisher","first-page":"110","DOI":"10.1016\/j.chemosphere.2014.09.033","article-title":"A 17-fold\nincrease of trifluoroacetic acid in landscape\nwaters of Beijing, China during the last decade","volume":"129","author":"Zhai","year":"2015","journal-title":"Chemosphere"},{"issue":"49","key":"2026092909071781900_cit11","doi-asserted-by":"publisher","first-page":"26762","DOI":"10.1021\/acs.est.5c10868","article-title":"Tracking Trifluoroacetate (TFA) through\nTime: A 78-Year Record from Archived Wines","volume":"59","author":"Freeling","year":"2025","journal-title":"Environ. Sci. Technol."},{"key":"2026092909071781900_cit12","doi-asserted-by":"publisher","first-page":"460","DOI":"10.1016\/j.watres.2017.09.045","article-title":"Small, mobile, persistent:\nTrifluoroacetate in the water cycle - Overlooked sources, pathways,\nand consequences for drinking water supply","volume":"126","author":"Scheurer","year":"2017","journal-title":"Water Res."},{"key":"2026092909071781900_cit13","unstructured":"Futures, G.\n          \n          Contamination\ndes eaux par des \u201cpolluants e\u0301ternels\u201d a\u0300\nSalindres, 2024."},{"key":"2026092909071781900_cit14","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2019.105295","article-title":"Distribution of novel\nand legacy per-\/polyfluoroalkyl substances in serum and its associations\nwith two glycemic biomarkers among Chinese adult men and women with\nnormal blood glucose levels","volume":"134","author":"Duan","year":"2020","journal-title":"Environ. Int."},{"issue":"42","key":"2026092909071781900_cit15","doi-asserted-by":"publisher","first-page":"15782","DOI":"10.1021\/acs.est.2c06715","article-title":"Elevated Levels\nof Ultrashort- and\nShort-Chain Perfluoroalkyl Acids in US Homes and People","volume":"57","author":"Zheng","year":"2023","journal-title":"Environ. Sci. Technol."},{"issue":"4","key":"2026092909071781900_cit16","doi-asserted-by":"publisher","first-page":"1069","DOI":"10.1007\/s00204-023-03454-y","article-title":"Mammalian toxicity\nof trifluoroacetate and assessment\nof human health risks due to environmental exposures","volume":"97","author":"Dekant","year":"2023","journal-title":"Arch. Toxicol."},{"key":"2026092909071781900_cit17","unstructured":"Registration Dossier for Trifluoroacetic\nAcid; EC Number 200-929-3;\nCAS Number 76-05-1. https:\/\/echa.europa.eu\/de\/registration-dossier\/-\/registered-dossier\/5203\/7\/1."},{"key":"2026092909071781900_cit18","first-page":"1725","volume-title":"The Merck Index: An Encyclopedia of Chemicals,\nDrugs, and Biologicals","author":"O\u2019Neil","year":"2001","edition":"13"},{"issue":"3","key":"2026092909071781900_cit19","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1016\/j.tcb.2017.10.010","article-title":"Endothelial\nCell Metabolism in Health and Disease","volume":"28","author":"Rohlenova","year":"2018","journal-title":"Trends\nCell Biol."},{"issue":"12","key":"2026092909071781900_cit20","doi-asserted-by":"publisher","first-page":"923","DOI":"10.1038\/s41569-025-01162-x","article-title":"Endothelial cell metabolism in cardiovascular physiology\nand disease","volume":"22","author":"Pasut","year":"2025","journal-title":"Nat. Rev. Cardiol."},{"issue":"2","key":"2026092909071781900_cit21","doi-asserted-by":"publisher","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: improving the speed and accuracy of\ndocking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"issue":"8","key":"2026092909071781900_cit22","doi-asserted-by":"publisher","first-page":"3891","DOI":"10.1021\/acs.jcim.1c00203","article-title":"AutoDock Vina 1.2.0:\nNew Docking Methods, Expanded Force Field, and Python Bindings","volume":"61","author":"Eberhardt","year":"2021","journal-title":"J. Chem. Inf. Model."},{"issue":"2","key":"2026092909071781900_cit23","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1006\/dbio.2002.0711","article-title":"In vivo\nimaging of embryonic vascular development using\ntransgenic zebrafish","volume":"248","author":"Lawson","year":"2002","journal-title":"Dev. 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Diabetes Rep."},{"issue":"3","key":"2026092909071781900_cit26","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1016\/j.cell.2013.06.037","article-title":"Role of PFKFB3-driven glycolysis in vessel sprouting","volume":"154","author":"De Bock","year":"2013","journal-title":"Cell"},{"issue":"7546","key":"2026092909071781900_cit27","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1038\/nature14362","article-title":"Fatty acid\ncarbon is essential for dNTP synthesis in endothelial cells","volume":"520","author":"Schoors","year":"2015","journal-title":"Nature"},{"issue":"5","key":"2026092909071781900_cit28","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1038\/s42255-021-00390-y","article-title":"Single-cell\nmetabolic imaging reveals\na SLC2A3-dependent glycolytic burst in motile endothelial cells","volume":"3","author":"Wu","year":"2021","journal-title":"Nat. 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Adv."},{"key":"2026092909071781900_cit32","volume-title":"Lehninger Principles\nof Biochemistry","author":"Nelson","year":"2021","edition":"8"},{"issue":"2","key":"2026092909071781900_cit33","doi-asserted-by":"publisher","first-page":"57","DOI":"10.32607\/20758251-2015-7-2-57-63","article-title":"Search for Human Lactate Dehydrogenase\nA Inhibitors Using Structure-Based Modeling","volume":"7","author":"Nilov","year":"2015","journal-title":"Acta Nat."},{"issue":"Pt 5","key":"2026092909071781900_cit34","doi-asserted-by":"publisher","first-page":"1484","DOI":"10.1107\/S1399004714005422","article-title":"Structural characterization\nof the apo form and NADH binary complex of human lactate dehydrogenase","volume":"70","author":"Dempster","year":"2014","journal-title":"Acta Crystallogr., Sect. D: Biol. Crystallogr."},{"issue":"5","key":"2026092909071781900_cit35","doi-asserted-by":"publisher","first-page":"1193","DOI":"10.1111\/php.12775","article-title":"Mechanistic\nAnalysis of Fluorescence Quenching of Reduced\nNicotinamide Adenine Dinucleotide by Oxamate in Lactate Dehydrogenase\nTernary Complexes","volume":"93","author":"Peng","year":"2017","journal-title":"Photochem. Photobiol"},{"issue":"12","key":"2026092909071781900_cit36","doi-asserted-by":"publisher","DOI":"10.3390\/molecules22122217","article-title":"Three-Dimensional\nAnalysis of the Interactions between hLDH5 and Its Inhibitors","volume":"22","author":"Poli","year":"2017","journal-title":"Molecules"},{"issue":"1","key":"2026092909071781900_cit37","doi-asserted-by":"publisher","first-page":"214","DOI":"10.1038\/s42003-023-04579-7","article-title":"RhoGDI\u03b1 regulates\nspermatogenesis through Rac1\/cofilin\/F-actin\nsignaling","volume":"6","author":"Zhu","year":"2023","journal-title":"Commun. 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U.S.A."},{"issue":"6527","key":"2026092909071781900_cit40","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1126\/science.abb2683","article-title":"Glycolysis\nfuels phosphoinositide 3-kinase signaling to bolster T cell immunity","volume":"371","author":"Xu","year":"2021","journal-title":"Science"},{"key":"2026092909071781900_cit41","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2023.216285","article-title":"NUSAP1-LDHA-Glycolysis-Lactate\nfeedforward loop promotes Warburg effect and metastasis in pancreatic\nductal adenocarcinoma","volume":"567","author":"Chen","year":"2023","journal-title":"Cancer Lett."},{"issue":"5","key":"2026092909071781900_cit42","doi-asserted-by":"publisher","DOI":"10.1289\/EHP13214","article-title":"Development\nof an Automated Morphometric Approach to Assess Vascular Outcomes\nfollowing Exposure to Environmental Chemicals in Zebrafish","volume":"132","author":"Zhong","year":"2024","journal-title":"Environ. Health Perspect."},{"issue":"25","key":"2026092909071781900_cit43","doi-asserted-by":"publisher","first-page":"12495","DOI":"10.1021\/acs.est.4c14490","article-title":"Small Black\nPhosphorus Disrupts Vascular Development and Hematopoiesis in Zebrafish\nLarvae","volume":"59","author":"Zhang","year":"2025","journal-title":"Environ. Sci. Technol."},{"issue":"48","key":"2026092909071781900_cit44","doi-asserted-by":"publisher","first-page":"26041","DOI":"10.1021\/acs.est.5c11430","article-title":"Identification of Key\nComponents with Cardiovascular Effect in Airborne Particulate Matter\nUsing an Expanded Component-separation Strategy","volume":"59","author":"Zhang","year":"2025","journal-title":"Environ. Sci. Technol."},{"issue":"1","key":"2026092909071781900_cit45","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1186\/s13040-025-00478-1","article-title":"A graph-theoretic\nframework for quantitative\nanalysis of angiogenic networks","volume":"18","author":"Okoro","year":"2025","journal-title":"BioData Min."},{"issue":"10","key":"2026092909071781900_cit46","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1000549","article-title":"Tipping the balance:\nrobustness of tip cell selection, migration and fusion in angiogenesis","volume":"5","author":"Bentley","year":"2009","journal-title":"PLoS Comput. 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