Objective To analyze the potential impact of acetyl tributyl citrate (ATBC), a commonly used plasticizer substitute, on the pathogenesis of Alzheimer's disease (AD), and to reveal the toxic effects of ATBC on AD and the complex interactions between key molecular pathways.

Methods The toxicological characteristics of ATBC were predicted using the ADMETlab2.0 and ProTox3.0 platforms. The targets of ATBC were screened using the SwissTargetPrediction and ChEMBL databases. AD-related targets were collected from the GeneCards and OMIM databases.A protein-protein interaction network was constructed to identify the core targets, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses were performed. Finally, molecular docking was used to verify the binding affinity between key molecules.

Results The toxicity prediction results showed that ATBC has neurotoxicity such as blood-brain barrier damage. 124 intersection targets of ATBC and AD were screened out, among which STAT3, EGFR, MMP9, CTSB and MAPK1 were the core targets. The functional enrichment analysis results showed that ATBC mainly affects the development of AD by interfering with key biological processes such as inflammatory response, GABAergic neurotransmission, amyloid protein metabolism and protein kinase signaling. The molecular docking results confirmed that ATBC has strong binding affinity with five core targets (binding energies was less than -5.0 kcal/mol), with the most stable binding to STAT3 (-7.1 kcal/mol).

Conclusion ATBC can promote the pathological process of AD through multiple synergistic mechanisms, mainly manifested in its high affinity binding to key targets such as STAT3, EGFR, MMP9, CTSB, and MAPK1, thereby activating neuroinflammation response, disrupting GABAergic synaptic transmission, inducing insulin resistance and metabolic disorders in the brain, and accelerating amyloid β-protein deposition and tau protein pathological processes. These findings provide an important theoretical basis for assessing the neurotoxicity risk of ATBC and reference for developing environmental control strategies and multi-target intervention programs for AD.

1. 王亚楠, 任凌, 韩伟, 等. 阿尔茨海默病病理生理机制及其诊断研究进展[J]. 解放军医学杂志, 2025, 50(5): 632-640.WangYN, RenL, HanW, et al. Advances in research of pathophysiological mechanisms and diagnosis of Alzheimer's disease[J]. Medical Journal of Chinese People's Liberation Army, 2025, 50(5): 632-640. DOI: 10.11855/j.issn.0577-7402.0467.2024.0913.

2. 王刚, 齐金蕾, 刘馨雅, 等. 中国阿尔茨海默病报告2024[J]. 诊断学理论与实践, 2024, 23(3): 219-256.WangG, QiJL, LiuXY, et al. China Alzheimer Report 2024[J]. Journal of Diagnostics Concepts & Practice, 2024, 23(3): 219-256. DOI: 10.16150/j.1671-2870.2024.03.001.

3. ChenS, CaoZ, NandiA, et al. The global macroeconomic burden of Alzheimer's disease and other dementias: estimates and projections for 152 countries or territories[J]. Lancet Glob Health, 2024, 12(9): e1534-e1543. DOI: 10.1016/s2214-109x(24)00264-x.

4. LiuE, ZhangY, WangJZ. Updates in Alzheimer's disease: from basic research to diagnosis and therapies[J]. Transl Neurodegener, 2024, 13(1): 45. DOI: 10.1186/s40035-024-00432-x.

5. ParumsDV. A review of the current status of disease-modifying therapies and prevention of Alzheimer's disease[J]. Med Sci Monit, 2024, 30: e945091. DOI: 10.12659/msm.945091.

6. 王婷, 高星梅, 郭会彩, 等. 增塑剂DEHP神经毒性作用及机制的研究进展[J]. 生态毒理学报, 2022, 17(2): 33-42.WangT, GaoXM, GuoHC, et al. Research progress on neurotoxic effects and mechanism of plasticizer DEHP[J]. Asian Journal of Ecotoxicology, 2022, 17(2): 33-42. DOI: 10.7524/AJE.1673-5897.20210503001.

7. 张亚运. 新型塑化剂柠檬酸酯CAEs的污染特征、生物转化及脂质代谢干扰研究[D]. 南京: 南京理工大学, 2022.ZhangYY. Occurrence, biotransformation,and lipidmetabolism disturbance of a class of novelplasticizers—citric acid esters[D]. Nanjing: Nanjing University of Science and Technology, 2022. DOI: 10.27241/d.cnki.gnjgu.2022.000106.

8. MalarvannanG, OnghenaM, VerstraeteS, et al. Phthalate and alternative plasticizers in indwelling medical devices in pediatric intensive care units[J]. J Hazard Mater, 2019, 363: 64-72. DOI: 10.1016/j.jhazmat.2018.09.087.

9. JeonH, YeoS, ParkEA, et al. Identification and quantification of acetyl tributyl citrate (ATBC) metabolites using human liver microsomes and human urine[J]. Chemosphere, 2024, 363: 142840. DOI: 10.1016/j.chemosphere.2024.142840.

10. ZhangY, ZhangZ, ZhuS, et al. Acetyl tributyl citrate exposure at seemingly safe concentrations induces adverse effects in different genders of type 2 diabetes mice, especially brain tissue[J]. Toxics, 2023, 11(10): 877. DOI: 10.3390/toxics11100877.

11. HuangJ, ZengL, YangQ,et al. Integrated multi-omics analysis identifies TLR4-mediated mechanisms in ATBC-induced ovarian dysfunction and female infertility: a network toxicology, transcriptomic, and Mendelian randomization study[J]. J Ovarian Res, 2025, 18(1): 120. DOI: 10.1186/s13048-025-01708-0.

12. YunK, JeonH, LeeJ, et al. Effects of two alternative plasticizers on the growth hormone-related endocrine system, neurodevelopment, and oxidative stress of zebrafish larvae[J]. Environ Pollut, 2024, 341: 122947. DOI: 10.1016/j.envpol.2023.122947.

13. HorieY, YapCK, OkamuraH. Developmental toxicity and thyroid hormone-disrupting effects of acetyl tributyl citrate in zebrafish and Japanese medaka[J]. Journal of Hazardous Materials Advances, 2022, 8: 100199. DOI: 10.1016/j.hazadv.2022.100199.

14. GuoDQ, FangYM, SunZD, et al. Analyzing the potential targets and mechanisms of liver damage induced by acetyl tributyl citrate plasticizer using network toxicology, molecular docking and in vitro experiments[J]. Front Pharmacol, 2025, 16: 1636576. DOI: 10.3389/fphar.2025.1636576.

15. Del GiudiceG, SerraA, PavelA, et al. A network toxicology approach for mechanistic modelling of nanomaterial hazard and adverse outcomes[J]. Adv Sci (Weinh), 2024, 11(32): e2400389. DOI: 10.1002/advs.202400389.

16. QuT, SunQ, TanB, et al. Integration of network toxicology and transcriptomics reveals the novel neurotoxic mechanisms of 2, 2', 4, 4'-tetrabromodiphenyl ether[J]. J Hazard Mater, 2025, 486: 136999. DOI: 10.1016/j.jhazmat.2024.136999.

17. LiB, XuC, ZhangD, et al. Combined analysis of network toxicology and multiomics revealed the potential mechanism of 6PPDQ-induced hepatotoxicity in mice[J]. Environ Sci Technol, 2025, 59(21): 10204-10214. DOI: 10.1021/acs.est.5c03906.

18. KitchenDB, DecornezH, FurrJR, et al. Docking and scoring in virtual screening for drug discovery: methods and applications[J]. Nat Rev Drug Discov, 2004, 3(11): 935-949. DOI: 10.1038/nrd1549.

19. TaoQQ, LinRR, ChenYH, et al. Discerning the role of blood brain barrier dysfunction in Alzheimer's disease[J]. Aging Dis, 2022, 13(5): 1391-1404. DOI: 10.14336/ad.2022.0130-1.

20. HuangS. Efficient analysis of toxicity and mechanisms of environmental pollutants with network toxicology and molecular docking strategy: acetyl tributyl citrate as an example[J]. Sci Total Environ, 2023, 905: 167904. DOI: 10.1016/j.scitotenv.2023.167904.

21. ZhangW, JieJ, XuQ, et al. Characterizing the obesogenic and fatty liver-inducing effects of acetyl tributyl citrate (ATBC) plasticizer using both in vivo and in vitro models[J]. J Hazard Mater, 2023, 445: 130548. DOI: 10.1016/j.jhazmat.2022.130548.

22. ZimmermannL, DierkesG, TernesTA, et al. Benchmarking the in vitro toxicity and chemical composition of plastic consumer products[J]. Environ Sci Technol, 2019, 53(19): 11467-11477. DOI: 10.1021/acs.est.9b02293.

23. SackmannK, ReemtsmaT, RahmbergM, et al. Impact of European chemicals regulation on the industrial use of plasticizers and patterns of substitution in Scandinavia[J]. Environ Int, 2018, 119: 346-352. DOI: 10.1016/j.envint.2018.06.037.

24. Serrano-RegalMP, Bayón-CorderoL, OrdazRP, et al. Expression and function of GABA receptors in myelinating cells[J]. Front Cell Neurosci, 2020, 14: 256. DOI: 10.3389/fncel.2020.00256.

25. CastellanoD, ShepardRD, LuW. Looking for novelty in an "old" receptor: recent advances toward our understanding of GABA_ARs and their implications in receptor pharmacology[J]. Front Neurosci, 2021, 14: 616298. DOI: 10.3389/fnins.2020.616298.

26. WenY, DongZ, LiuJ, et al. Glutamate and GABA(A) receptor crosstalk mediates homeostatic regulation of neuronal excitation in the mammalian brain[J]. Signal Transduct Target Ther, 2022, 7(1): 340. DOI: 10.1038/s41392-022-01148-y.

27. KelleherRJ3rd, GovindarajanA, JungHY, et al. Translational control by MAPK signaling in long-term synaptic plasticity and memory[J]. Cell, 2004, 116(3): 467-479. DOI: 10.1016/s0092-8674(04)00115-1.

28. OdorčićI, HamedMB, LismontS, et al. Apo and Aβ46-bound γ-secretase structures provide insights into amyloid-β processing by the APH-1B isoform[J]. Nat Commun, 2024, 15(1): 4479. DOI: 10.1038/s41467-024-48776-2.

29. DuanT, DuY, XingC, et al. Toll-like receptor signaling and its role in cell-mediated immunity[J]. Front Immunol, 2022, 13: 812774. DOI: 10.3389/fimmu.2022.812774.

30. AitchisonEE, DimesaAM, ShoariA. Matrix metalloproteinases in glioma: drivers of invasion and therapeutic targets[J]. BioTech (Basel), 2025, 14(2): 28. DOI: 10.3390/biotech14020028.

31. CoppiE, DettoriI, CherchiF, et al. New insight into the role of adenosine in demyelination, stroke and neuropathic pain[J]. Front Pharmacol, 2021, 11: 625662. DOI: 10.3389/fphar.2020.625662.

32. MihaylovaMM, VasquezDS, RavnskjaerK, et al. Class IIa histone deacetylases are hormone-activated regulators of FOXO and mammalian glucose homeostasis[J]. Cell, 2011, 145(4): 607-621. DOI: 10.1016/j.cell.2011.03.043.

33. WenX, HuJ. Targeting STAT3 signaling pathway in the treatment of Alzheimer's disease with compounds from natural products[J]. Int Immunopharmacol, 2024, 141: 112936. DOI: 10.1016/j.intimp.2024.112936.

34. ThakurS, DhapolaR, SarmaP, et al. Neuroinflammation in Alzheimer's disease: current progress in molecular signaling and therapeutics[J]. Inflammation, 2023, 46(1): 1-17. DOI: 10.1007/s10753-022-01721-1.

35. ReichenbachN, DelekateA, PlescherM, et al. Inhibition of Stat3-mediated astrogliosis ameliorates pathology in an Alzheimer's disease model[J]. EMBO Mol Med, 2019, 11(2): e9665. DOI: 10.15252/emmm.201809665.

36. JayaswamyPK, VijaykrishnarajM, PatilP, et al. Implicative role of epidermal growth factor receptor and its associated signaling partners in the pathogenesis of Alzheimer's disease[J]. Ageing Res Rev, 2023, 83: 101791. DOI: 10.1016/j.arr.2022.101791.

37. RajmohanR, ReddyPH. Amyloid-beta and phosphorylated tau accumulations cause abnormalities at synapses of Alzheimer's disease neurons[J]. J Alzheimers Dis, 2017, 57(4): 975-999. DOI: 10.3233/jad-160612.

38. LimNK, VillemagneVL, SoonCP, et al. Investigation of matrix metalloproteinases, MMP-2 and MMP-9, in plasma reveals a decrease of MMP-2 in Alzheimer's disease[J]. J Alzheimers Dis, 2011, 26(4): 779-786. DOI: 10.3233/jad-2011-101974.

39. SongJ, WuC, KorposE, et al. Focal MMP-2 and MMP-9 activity at the blood-brain barrier promotes chemokine-induced leukocyte migration[J]. Cell Rep, 2015, 10(7): 1040-1054. DOI: 10.1016/j.celrep.2015.01.037.

40. SunY, RongX, LuW, et al. Translational study of Alzheimer's disease (AD) biomarkers from brain tissues in AβPP/PS1 mice and serum of AD patients[J]. J Alzheimers Dis, 2015, 45(1): 269-282. DOI: 10.3233/jad-142805.

41. RawatP, SeharU, BishtJ, et al. Phosphorylated tau in Alzheimer's disease and other tauopathies[J]. Int J Mol Sci, 2022, 23(21): 12841. DOI: 10.3390/ijms232112841.

42. FerrerI, BlancoR, CarmonaM, et al. Phosphorylated map kinase (ERK1, ERK2) expression is associated with early tau deposition in neurones and glial cells, but not with increased nuclear DNA vulnerability and cell death, in Alzheimer disease, Pick's disease, progressive supranuclear palsy and corticobasal degeneration[J]. Brain Pathol, 2001, 11(2): 144-158. DOI: 10.1111/j.1750-3639.2001.tb00387.x.

43. XiaY, ProkopS, GiassonBI. "Don't Phos Over Tau": recent developments in clinical biomarkers and therapies targeting tau phosphorylation in Alzheimer's disease and other tauopathies[J]. Mol Neurodegener, 2021, 16(1): 37. DOI: 10.1186/s13024-021-00460-5.

44. XuY, ZhaoM, HanY, et al. GABAergic inhibitory interneuron deficits in Alzheimer's disease: implications for treatment[J]. Front Neurosci, 2020, 14: 660. DOI: 10.3389/fnins.2020.00660.

45. LiY, SunH, ChenZ, et al. Implications of GABAergic neurotransmission in Alzheimer's disease[J]. Front Aging Neurosci, 2016, 8: 31. DOI: 10.3389/fnagi.2016.00031.

46. RosenbergerAF, HilhorstR, CoartE, et al. Protein kinase activity decreases with higher braak stages of Alzheimer's disease pathology[J]. J Alzheimers Dis, 2016, 49(4): 927-943. DOI: 10.3233/jad-150429.

47. HouJ, ChenY, Grajales-ReyesG, et al. TREM2 dependent and independent functions of microglia in Alzheimer's disease[J]. Mol Neurodegener, 2022, 17(1): 84. DOI: 10.1186/s13024-022-00588-y.

48. HurJY, FrostGR, WuX, et al. The innate immunity protein IFITM3 modulates γ-secretase in Alzheimer's disease[J]. Nature, 2020, 586(7831): 735-740. DOI: 10.1038/s41586-020-2681-2.

49. Ferreira-VieiraTH, GuimaraesIM, SilvaFR, et al. Alzheimer's disease: targeting the cholinergic system[J]. Curr Neuropharmacol, 2016, 14(1): 101-115. DOI: 10.2174/1570159x13666150716165726.

50. SunB, HalabiskyB, ZhouY, et al. Imbalance between GABAergic and glutamatergic transmission impairs adult neurogenesis in an animal model of Alzheimer's disease[J]. Cell Stem Cell, 2009, 5(6): 624-633. DOI: 10.1016/j.stem.2009.10.003.

51. BasavarajappaBS, ShivakumarM, JoshiV, et al. Endocannabinoid system in neurodegenerative disorders[J]. J Neurochem, 2017, 142(5): 624-648. DOI: 10.1111/jnc.14098.

52. AsoE, FerrerI. CB2 cannabinoid receptor as potential target against Alzheimer's disease[J]. Front Neurosci, 2016, 10: 243. DOI: 10.3389/fnins.2016.00243.

53. ArnoldSE, ArvanitakisZ, Macauley-RambachSL, et al. Brain insulin resistance in type 2 diabetes and Alzheimer disease: concepts and conundrums[J]. Nat Rev Neurol, 2018, 14(3): 168-181. DOI: 10.1038/nrneurol.2017.185.

54. KellarD, CraftS. Brain insulin resistance in Alzheimer's disease and related disorders: mechanisms and therapeutic approaches[J]. Lancet Neurol, 2020, 19(9): 758-766. DOI: 10.1016/s1474-4422(20)30231-3.

55. HijaziS, HeistekTS, ScheltensP, et al. Early restoration of parvalbumin interneuron activity prevents memory loss and network hyperexcitability in a mouse model of Alzheimer's disease[J]. Mol Psychiatry, 2020, 25(12): 3380-3398. DOI: 10.1038/s41380-019-0483-4.

56. LiW, ChenQ, PengC, et al. Roles of the receptor for advanced glycation end products and its ligands in the pathogenesis of Alzheimer's disease[J]. Int J Mol Sci, 2025, 26(1): 403. DOI: 10.3390/ijms26010403.

57. BenedictC, GrilloCA. Insulin resistance as a therapeutic target in the treatment of Alzheimer's disease: a state-of-the-art review[J]. Front Neurosci, 2018, 12: 215. DOI: 10.3389/fnins.2018.00215.

58. SędzikowskaA, SzablewskiL. Insulin and insulin resistance in Alzheimer's disease[J]. Int J Mol Sci, 2021, 22(18): 9987. DOI: 10.3390/ijms22189987.

59. KongY, LiuC, ZhouY, et al. Progress of RAGE molecular imaging in Alzheimer's disease[J]. Front Aging Neurosci, 2020, 12: 227. DOI: 10.3389/fnagi.2020.00227.

60. YanSD, BierhausA, NawrothPP, et al. RAGE and Alzheimer's disease: a progression factor for amyloid-beta-induced cellular perturbation?[J]. J Alzheimers Dis, 2009, 16(4): 833-843. DOI: 10.3233/jad-2009-1030.