{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T11:26:37Z","timestamp":1762255597387,"version":"build-2065373602"},"reference-count":44,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T00:00:00Z","timestamp":1762128000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ningxia Science Foundation of China","award":["2024AAC03222"],"award-info":[{"award-number":["2024AAC03222"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82160608"],"award-info":[{"award-number":["82160608"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ningxia Medical University Scientific Research Project","award":["XT2022007"],"award-info":[{"award-number":["XT2022007"]}]},{"name":"Innovation and Entrepreneurship Training Program for College Students at Ningxia Medical University","award":["X202410752093"],"award-info":[{"award-number":["X202410752093"]}]},{"name":"Ningxia Medical University Distinctive Disciplines","award":["TSXK2025002"],"award-info":[{"award-number":["TSXK2025002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>2,4-Dichlorophenoxyacetic acid (2,4-D) is a widely used herbicide, yet its potential to induce hepatorenal injury via oxidative stress and apoptosis raises significant health concerns. Lycium barbarum polysaccharides (LBP) possess recognized antioxidant and anti-apoptotic properties, but their protective mechanisms against 2,4-D toxicity, particularly through a multi-target network, remain inadequately explored. This study aimed to systematically investigate the mechanisms of 2,4-D-induced hepatorenal injury and the protective efficacy of LBP by integrating network toxicology, molecular docking, and experimental validation. An integrated approach was employed. Core targets and pathways were identified via network toxicology. Molecular docking predicted interactions between 2,4-D and these targets. In vivo validation was conducted on Sprague-Dawley rats treated with 2,4-D (75 mg\/kg) and\/or LBP (50 mg\/kg) for 28 days, assessing histopathology, serum oxidative stress markers superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and cellular apoptosis (TUNEL staining). Network analysis identified PPARG, NFKB1, PPARA, NFE2L2, and SERPINE1 as core targets, with molecular docking confirming strong binding affinities (binding energies: \u22125.1 to \u22126.3 kcal\u00b7mol\u22121) and KEGG enrichment implicating cAMP, Ca2+, and PPAR signaling pathways. Experimentally, 2,4-D exposure induced significant histopathological damage, suppressed SOD\/GSH-Px activities (p &lt; 0.001), elevated MDA levels (p &lt; 0.001), and markedly increased renal apoptosis (p &lt; 0.01). Crucially, LBP intervention substantially mitigated these alterations, ameliorating tissue injury, restoring antioxidant defenses, increasing SOD\/GSH-Px (p &lt; 0.01), reducing MDA (p &lt; 0.001) and significantly decreasing renal apoptosis (p &lt; 0.05). This study elucidates a multi-target mechanism for 2,4-D-induced hepatorenal injury centered on oxidative stress\u2013apoptosis dysregulation and demonstrates that LBP confers significant protection likely via modulation of this network. These findings underscore the potential of LBP as a natural protective agent against pesticide-induced organ damage and highlight the utility of integrated network approaches in toxicological research.<\/jats:p>","DOI":"10.3390\/ijms262110685","type":"journal-article","created":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:15:14Z","timestamp":1762175714000},"page":"10685","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Elucidating the Mechanism of Liver and Kidney Damage in Rats Caused by Exposure to 2,4-Dichlorophenoxyacetic Acid and the Protective Effect of Lycium barbarum Polysaccharides Based on Network Toxicology and Molecular Docking"],"prefix":"10.3390","volume":"26","author":[{"given":"Xiaoqi","family":"Luo","sequence":"first","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yixuan","family":"Wei","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinyu","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoning","family":"Meng","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yating","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Na","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huifang","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7619-0228","authenticated-orcid":false,"given":"Jian","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China"},{"name":"Key Laboratory of Environmental Factors and Chronic Disease Control, School of Public Health, Ningxia Medical University, Yinchuan 750004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1177\/0049475518786834","article-title":"2,4-D poisoning: A review with illustration of two cases","volume":"48","author":"Pannu","year":"2018","journal-title":"Trop. Doct."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.tibtech.2007.06.002","article-title":"Environmental genomics: Mechanistic insights into toxicity of and resistance to the herbicide 2,4-D","volume":"25","author":"Teixeira","year":"2007","journal-title":"Trends Biotechnol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"104783","DOI":"10.1016\/j.tiv.2020.104783","article-title":"Cytotoxic and genotoxic assessments of 2,4-dichlorophenoxyacetic acid (2,4-D) in in vitro mammalian cells","volume":"65","author":"Laborde","year":"2020","journal-title":"Toxicol. Vitr."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Tian, X., Liang, T., Liu, Y., Ding, G., Zhang, F., and Ma, Z. (2019). Extraction, Structural Characterization, and Biological Functions of Lycium Barbarum Polysaccharides: A Review. Biomolecules, 9.","DOI":"10.3390\/biom9090389"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"125016","DOI":"10.1016\/j.chemosphere.2019.125016","article-title":"Analysis of global trends and gaps for studies about 2,4-D herbicide toxicity: A scientometric review","volume":"241","author":"Zuanazzi","year":"2020","journal-title":"Chemosphere"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2454","DOI":"10.1002\/tox.23358","article-title":"Effects of 2,4-dichlorophenoxyacetic acid on the expression of NLRP3 inflammasome and autophagy-related proteins as well as the protective effect of Lycium barbarum polysaccharide in neonatal rats","volume":"36","author":"Zhou","year":"2021","journal-title":"Environ. Toxicol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e45124","DOI":"10.15252\/embr.201745124","article-title":"SIRT5 inhibits peroxisomal ACOX1 to prevent oxidative damage and is downregulated in liver cancer","volume":"19","author":"Chen","year":"2018","journal-title":"EMBO Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1681\/ASN.2015121308","article-title":"Renal Aging: Causes and Consequences","volume":"28","author":"Hughes","year":"2017","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1038\/s41571-020-0341-y","article-title":"Targeting apoptosis in cancer therapy","volume":"17","author":"Carneiro","year":"2020","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"117197","DOI":"10.1016\/j.jep.2023.117197","article-title":"Intervention effect of Lycium barbarum polysaccharide on lead-induced kidney injury mice and its mechanism: A study based on the PI3K\/Akt\/mTOR signaling pathway","volume":"319","author":"Xie","year":"2024","journal-title":"J. Ethnopharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"102578","DOI":"10.1016\/j.clinre.2025.102578","article-title":"Research progress on the regulatory role of different cell death pathways in metabolic-dysfunction-associated steatotic liver disease","volume":"49","author":"Zhang","year":"2025","journal-title":"Clin. Res. Hepatol. Gastroenterol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1152\/physrev.00061.2009","article-title":"Hepatocyte death: A clear and present danger","volume":"90","author":"Malhi","year":"2010","journal-title":"Physiol. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1038\/ki.2011.120","article-title":"Apoptosis and acute kidney injury","volume":"80","author":"Havasi","year":"2011","journal-title":"Kidney Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e12829","DOI":"10.1111\/cpr.12829","article-title":"TNF-\u03b1\/HMGB1 inflammation signalling pathway regulates pyroptosis during liver failure and acute kidney injury","volume":"53","author":"Wang","year":"2020","journal-title":"Cell Prolif."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Peng, J., Wang, L., Wang, M., Du, R., Qin, S., Jin, C.-Y., and Wei, Y. (2021). Yeast Synthetic Biology for the Production of Lycium barbarum Polysaccharides. Molecules, 26.","DOI":"10.3390\/molecules26061641"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1080\/13510002.2022.2036507","article-title":"Lycium Barbarum polysaccharide protects HaCaT cells from PM2.5-induced apoptosis via inhibiting oxidative stress, ER stress and autophagy","volume":"27","author":"Zhu","year":"2022","journal-title":"Redox Rep."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Yang, Y., Yu, L., Zhu, T., Xu, S., He, J., Mao, N., Liu, Z., and Wang, D. (2023). Neuroprotective effects of Lycium barbarum polysaccharide on light-induced oxidative stress and mitochondrial damage via the Nrf2\/HO-1 pathway in mouse hippocampal neurons. Int. J. Biol. Macromol., 251.","DOI":"10.1016\/j.ijbiomac.2023.126315"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1016\/0006-2952(68)90363-8","article-title":"Enhanced biliary excretion of thyroxine glucuronide in rats pretreated with benzpyrene","volume":"17","author":"Goldstein","year":"1968","journal-title":"Biochem. Pharmacol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"153847","DOI":"10.1016\/j.tox.2024.153847","article-title":"Analysis of environmental pollutant Bisphenol F elicited prostate injury targets and underlying mechanisms through network toxicology, molecular docking, and multi-level bioinformatics data integration","volume":"506","author":"Huang","year":"2024","journal-title":"Toxicology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.drudis.2021.09.007","article-title":"Machine-learning methods for ligand-protein molecular docking","volume":"27","author":"Crampon","year":"2022","journal-title":"Drug Discov. Today"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.theriogenology.2024.10.010","article-title":"Effects of PPARG on the proliferation, apoptosis, and estrogen secretion in goat granulosa cells","volume":"231","author":"Zhao","year":"2025","journal-title":"Theriogenology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1186\/s13018-023-03990-4","article-title":"Circ-NFKB1 sponges miR-203a-5p to regulate ERBB4 expression and promotes IL-1\u03b2 induced chondrocytes apoptosis","volume":"18","author":"Wang","year":"2023","journal-title":"J. Orthop. Surg. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"114271","DOI":"10.1016\/j.yexcr.2024.114271","article-title":"Long noncoding RNA UCA1 inhibits epirubicin-induced apoptosis by activating PPAR\u03b1-mediated lipid metabolism","volume":"442","author":"Sun","year":"2024","journal-title":"Exp. Cell Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"153466","DOI":"10.1016\/j.phymed.2021.153466","article-title":"Arbutin attenuates LPS-induced acute kidney injury by inhibiting inflammation and apoptosis via the PI3K\/Akt\/Nrf2 pathway","volume":"82","author":"Zhang","year":"2021","journal-title":"Phytomedicine"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"20241018","DOI":"10.1515\/med-2024-1018","article-title":"Sepsis induces the cardiomyocyte apoptosis and cardiac dysfunction through activation of YAP1\/Serpine1\/caspase-3 pathway","volume":"19","author":"Long","year":"2024","journal-title":"Open Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1017\/S2040174421000416","article-title":"Fetal programming by androgen excess impairs liver lipid content and PPARg expression in adult rats","volume":"13","author":"Silva","year":"2022","journal-title":"J. Dev. Orig. Health Dis."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"K\u00f6k\u00e9ny, G., Calvier, L., and Hansmann, G. (2021). PPAR\u03b3 and TGF\u03b2-Major Regulators of Metabolism, Inflammation, and Fibrosis in the Lungs and Kidneys. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms221910431"},{"key":"ref_28","first-page":"1011","article-title":"Ciliogenesis is reciprocally regulated by PPARA and NR1H4\/FXR through controlling autophagy in vitro and in vivo","volume":"14","author":"Liu","year":"2018","journal-title":"Autophagy"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1007\/s00280-021-04264-8","article-title":"Pharmacogenomics and functional imaging to predict irinotecan pharmacokinetics and pharmacodynamics: The predict IR study","volume":"88","author":"Michael","year":"2021","journal-title":"Cancer Chemother. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.1080\/15548627.2020.1712108","article-title":"SQSTM1\/p62 activates NFE2L2\/NRF2 via ULK1-mediated autophagic KEAP1 degradation and protects mouse liver from lipotoxicity","volume":"16","author":"Lee","year":"2020","journal-title":"Autophagy"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"114828","DOI":"10.1016\/j.intimp.2025.114828","article-title":"Targeting neutrophil extracellular traps: SERPINE1 and THBS1 as non-invasive biomarkers for early detection of liver fibrosis in metabolic dysfunction-associated Steatotic liver disease","volume":"158","author":"Yang","year":"2025","journal-title":"Int. Immunopharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Liu, R.J., Xu, Z.P., Li, S.Y., Yu, J.-J., Feng, N.-H., Xu, B., and Chen, M. (2022). BAP1-Related ceRNA (NEAT1\/miR-10a-5p\/SERPINE1) Promotes Proliferation and Migration of Kidney Cancer Cells. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.852515"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"38501","DOI":"10.1007\/s11356-020-10370-6","article-title":"Herbicides based on 2,4-D: Its behavior in agricultural environments and microbial biodegradation aspects. A review","volume":"27","author":"Magnoli","year":"2020","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Food Safety Commission of Japan (2017). 2,4-D (Pesticides). Food Saf., 5, 171\u2013177.","DOI":"10.14252\/foodsafetyfscj.2017010s"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1177\/0748233720983167","article-title":"2,4-Dichlorophenoxyacetic acid induced hepatic and renal toxicological perturbations in rat model: Attenuation by selenium supplementation","volume":"37","author":"Shafeeq","year":"2021","journal-title":"Toxicol. Ind. Health"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Martins, R.X., Vieira, L., Souza, J.A.C.R., Silva, M.G.F., Muniz, M.S., Souza, T., Queiroga, F.R., Machado, M.R.F., da Silva, P.M., and Farias, D. (2021). Exposure to 2,4-D herbicide induces hepatotoxicity in zebrafish larvae. Comp. Biochem. Physiol. C Toxicol. Pharmacol., 248.","DOI":"10.1016\/j.cbpc.2021.109110"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"26170","DOI":"10.1007\/s11356-019-05837-0","article-title":"2,4-D causes oxidative stress induction and apoptosis in human dental pulp stem cells (hDPSCs)","volume":"26","author":"Mahmoudinia","year":"2019","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Allam, A., Abdeen, A., Devkota, H.P., Ibrahim, S.S., Youssef, G., Soliman, A., Abdel-Daim, M.M., Alzahrani, K.J., Shoghy, K., and Ibrahim, S.F. (2022). N-Acetylcysteine Alleviated the Deltamethrin-Induced Oxidative Cascade and Apoptosis in Liver and Kidney Tissues. Int. J. Environ. Res. Public. Health, 19.","DOI":"10.3390\/ijerph19020638"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.fct.2018.01.055","article-title":"Protective effects of rosuvastatin and vitamin E against fipronil-mediated oxidative damage and apoptosis in rat liver and kidney","volume":"114","author":"Abdeen","year":"2018","journal-title":"Food Chem. Toxicol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"23501","DOI":"10.1007\/s11356-020-11990-8","article-title":"Protective effect of Lycium barbarum polysaccharide on di-(2-ethylhexyl) phthalate-induced toxicity in rat liver","volume":"28","author":"Liu","year":"2021","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"110252","DOI":"10.1016\/j.cellsig.2022.110252","article-title":"Lycium barbarum polysaccharide antagonizes cardiomyocyte apoptosis by inhibiting the upregulation of GRK2 induced by I\/R injury, and salvage mitochondrial fission\/fusion imbalance and AKT\/eNOS signaling","volume":"92","author":"Li","year":"2022","journal-title":"Cell Signal."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Xie, W., Huang, Y.Y., Chen, H.G., and Zhou, X. (2021). Study on the Efficacy and Mechanism of Lycium barbarum Polysaccharide against Lead-Induced Renal Injury in Mice. Nutrients, 13.","DOI":"10.3390\/nu13092945"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhang, Z., Lai, G., and Sun, L. (2022). Basement-Membrane-Related Gene Signature Predicts Prognosis in WHO Grade II\/III Gliomas. Genes, 13.","DOI":"10.3390\/genes13101810"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1429","DOI":"10.18632\/aging.203886","article-title":"Identification and validation of an immune-related gene pairs signature for three urologic cancers","volume":"14","author":"Xie","year":"2022","journal-title":"Aging"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10685\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T11:14:29Z","timestamp":1762254869000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10685"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,3]]},"references-count":44,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262110685"],"URL":"https:\/\/doi.org\/10.3390\/ijms262110685","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2025,11,3]]}}}