{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T11:09:06Z","timestamp":1784977746341,"version":"3.55.0"},"reference-count":33,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T00:00:00Z","timestamp":1756771200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Open Foundation of Key Laboratory of Forensic Pathology, Ministry of Public Security, P.R. China","award":["GAFYBL202301"],"award-info":[{"award-number":["GAFYBL202301"]}]},{"name":"Open Foundation of Key Laboratory of Forensic Pathology, Ministry of Public Security, P.R. China","award":["SL2023A04J01627"],"award-info":[{"award-number":["SL2023A04J01627"]}]},{"name":"Science and Technology Program of Guangzhou, China","award":["GAFYBL202301"],"award-info":[{"award-number":["GAFYBL202301"]}]},{"name":"Science and Technology Program of Guangzhou, China","award":["SL2023A04J01627"],"award-info":[{"award-number":["SL2023A04J01627"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Xylazine abuse is emerging as a global problem, whereas the toxic mechanisms of xylazine poisoning are seldom studied. The present study aims to assess the heart injury in xylazine poisoning and uncover the underlying mechanism. Forty male SD rats were randomly dived into four groups: control (saline), low dose (10 mg\/kg xylazine), median dose (20 mg\/kg xylazine) and high dose (40 mg\/kg xylazine). The rats were injected with the drug intraperitoneally for 28 consecutive days, and then cardiac ultrasound examination was performed and serum and heart tissues were collected. Genomic, proteomic, and metabolic omics analyses were conducted. ELISA, RNA sequencing, histopathology examination, RT-qPCR, and Western blot were performed. Repeated injection of xylazine led to a decrease in the expression of cardiac output (CO), ventricular systole (VS), and ventricular diastole (VD), while concurrently elevating the levels of lactate dehydrogenase (LDH), creatine kinase myocardial band (CK-MB), and cardiac troponin T (c-TNT) in the serum. HE staining analysis showed evidence of contraction band necrosis, interstitial fibrosis, and infiltration by inflammatory cells in animals with xylazine poisoning. The modified genes, proteins, and metabolites were gathered, and the integration of transcriptomic, proteomic, and metabolic networks identified 25 overlapping pathways between the differentially expressed genes and metabolites (DEGs-DEMs) and the differentially expressed proteins and metabolites (DEPs-DEMs) joint pathways. The majority of these pathways pertained to the metabolism of sugars, amino acids, and fats. The proteins associated with fructose and mannose metabolism, as well as cholesterol metabolism, were validated, thereby substantiating their pivotal role in the development of xylazine-induced cardiac injury. Repeated injection of xylazine impaired heart function and the metabolism of fructose and mannose. Cholesterol metabolism pathways were critical in the process of xylazine-induced heart injury.<\/jats:p>","DOI":"10.3390\/ijms26178532","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T15:02:57Z","timestamp":1756825377000},"page":"8532","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Integrated Multilayer Omics Reveals the Underlying Mechanisms in Xylazine-Related Heart Injury in Rats"],"prefix":"10.3390","volume":"26","author":[{"given":"Yangchang","family":"Ou","sequence":"first","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Mai","sequence":"additional","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5987-2123","authenticated-orcid":false,"given":"Ning","family":"Wang","sequence":"additional","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyan","family":"Li","sequence":"additional","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yangchang","family":"Chen","sequence":"additional","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1584-712X","authenticated-orcid":false,"given":"Shuquan","family":"Zhao","sequence":"additional","affiliation":[{"name":"Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Smith, M.A., Biancorosso, S.L., Camp, J.D., Hailu, S.H., Johansen, A.N., Morris, M.H., and Carlson, H.N. (2023). \u201cTranq-dope\u201d overdose and mortality: Lethality induced by fentanyl and xylazine. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1280289"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1007\/s00213-023-06390-y","article-title":"Xylazine effects on opioid-induced brain hypoxia","volume":"240","author":"Choi","year":"2023","journal-title":"Psychopharmacology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1904","DOI":"10.1001\/jama.2023.8625","article-title":"Here\u2019s What to Know About Xylazine, aka Tranq, the Animal Tranquilizer Increasingly Found in Illicit Fentanyl Samples","volume":"329","author":"Rubin","year":"2023","journal-title":"JAMA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2209","DOI":"10.1056\/NEJMp2303120","article-title":"Xylazine\u2014Medical and Public Health Imperatives","volume":"388","author":"Gupta","year":"2023","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"102542","DOI":"10.1016\/j.jflm.2023.102542","article-title":"The first drug-related death associated with xylazine use in the UK and Europe","volume":"97","author":"Rock","year":"2023","journal-title":"J. Forensic Leg. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Yu, Z., Yuan, Y., Zhang, J., Gao, X., Chen, C., Zhao, E., Ma, J., and Zhou, Y. (2025). Forensic pathology and toxicological analyses: A case of fatal xylazine poisoning. J. Forensic Sci.","DOI":"10.1111\/1556-4029.70104"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Mai, T., Zhang, Y., and Zhao, S. (2023). Xylazine Poisoning in Clinical and Forensic Practice: Analysis Method, Characteristics, Mechanism and Future Challenges. Toxics, 11.","DOI":"10.3390\/toxics11121012"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.forsciint.2014.03.015","article-title":"Xylazine intoxication in humans and its importance as an emerging adulterant in abused drugs: A comprehensive review of the literature","volume":"240","year":"2014","journal-title":"Forensic Sci. Int."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7024","DOI":"10.1038\/s41467-022-34422-2","article-title":"Multi-omics signatures of the human early life exposome","volume":"13","author":"Maitre","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"156016","DOI":"10.1016\/j.phymed.2024.156016","article-title":"Multi-omics analysis provides new insights into mechanism of didymin on non-alcoholic fatty liver disease in rats","volume":"135","author":"Fang","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e023868","DOI":"10.1161\/JAHA.121.023868","article-title":"Integrated Multilayer Omics Reveals the Genomic, Proteomic, and Metabolic Influences of Histidyl Dipeptides on the Heart","volume":"11","author":"Yan","year":"2022","journal-title":"J. Am. Heart Assoc."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.jemermed.2015.09.051","article-title":"Xylazine Exposures Reported to Texas Poison Centers","volume":"51","author":"Forrester","year":"2016","journal-title":"J. Emerg. Med."},{"key":"ref_13","first-page":"78","article-title":"Xylazine was detected in a poisoning case","volume":"31","author":"Xu","year":"2023","journal-title":"Guang Dong Gong An Ke Ji"},{"key":"ref_14","first-page":"1540","article-title":"Detection of two food poisoning cases caused by xylazine residual using liquid chromatography-mass spectrometry","volume":"33","author":"Yao","year":"2023","journal-title":"Chin. J. Health Lab. Tec."},{"key":"ref_15","first-page":"12","article-title":"Effect of xylazine on isolated working heart in rat","volume":"20","author":"Dong","year":"1996","journal-title":"Bull. Acad. Mil. Med. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1161\/CIRCRESAHA.122.321369","article-title":"The Circadian Biology of Heart Failure","volume":"132","author":"Lordan","year":"2023","journal-title":"Circ. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1161\/CIRCULATIONAHA.121.056076","article-title":"Myocardial Rev-erb-Mediated Diurnal Metabolic Rhythm and Obesity Paradox","volume":"145","author":"Song","year":"2022","journal-title":"Circulation"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1161\/CIRCULATIONAHA.118.036550","article-title":"Circadian-Regulated Cell Death in Cardiovascular Diseases","volume":"139","author":"Lieberman","year":"2019","journal-title":"Circulation"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1905","DOI":"10.1093\/cvr\/cvad100","article-title":"Cardiac lipid metabolism, mitochondrial function, and heart failure","volume":"119","author":"Cabodevilla","year":"2023","journal-title":"Cardiovasc. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1093\/cvr\/cvx017","article-title":"Metabolism in cardiomyopathy: Every substrate matters","volume":"113","author":"Ritterhoff","year":"2017","journal-title":"Cardiovasc. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.cca.2021.05.034","article-title":"Identification of three novel pathogenic mutations in sarcomere genes associated with familial hypertrophic cardiomyopathy based on multi-omics study","volume":"520","author":"Liu","year":"2021","journal-title":"Clin. Chim. Acta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1016\/j.bbamcr.2011.01.015","article-title":"Targeting fatty acid and carbohydrate oxidation\u2014A novel therapeutic intervention in the ischemic and failing heart","volume":"1813","author":"Jaswal","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"E715","DOI":"10.1152\/ajpendo.00475.2015","article-title":"Measurement of \u03b2-oxidation capacity of biological samples by respirometry: A review of principles and substrates","volume":"310","author":"Ojuka","year":"2016","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Aitken-Buck, H.M., Krause, J., Zeller, T., Jones, P.P., and Lamberts, R.R. (2020). Long-Chain Acylcarnitines and Cardiac Excitation-Contraction Coupling: Links to Arrhythmias. Front. Physiol., 11.","DOI":"10.3389\/fphys.2020.577856"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wang, J., Zhang, F., Tan, C., Wang, N., Xia, X., Ye, Y., Cao, Y., Huang, X., Bai, Z., and Zhao, H. (2025). The VDAC3\/DHODH Axis Ameliorates Sepsis-induced Myocardial Injury by Regulating Ferroptosis. Front. Biosci., 30.","DOI":"10.31083\/FBL39559"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e00963","DOI":"10.1002\/advs.202500963","article-title":"PFKM-Driven Lactate Overproduction Promotes Atrial Fibrillation via Triggering Cardiac Fibroblasts Histone Lactylation","volume":"11","author":"Fang","year":"2025","journal-title":"Adv. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Yi, S., Mai, T., Fang, Y., Tian, Q., and Zhao, S. (2025). Repeated Injection of Xylazine Causes Liver Injury Through the PPAR Signaling Pathway in Rats. J. Biochem. Mol. Toxicol., 39.","DOI":"10.1002\/jbt.70101"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/gigascience\/gix120","article-title":"SOAPnuke: A MapReduce acceleration-supported software for integrated quality control and preprocessing of high-throughput sequencing data","volume":"7","author":"Chen","year":"2018","journal-title":"Gigascience"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.3317","article-title":"HISAT: A fast spliced aligner with low memory requirements","volume":"12","author":"Kim","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.1923","article-title":"Fast gapped-read alignment with Bowtie 2","volume":"9","author":"Langmead","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Li, B., and Dewey, C.N. (2011). RSEM: Accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform., 12.","DOI":"10.1186\/1471-2105-12-323"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1093\/bioinformatics\/btp612","article-title":"DEGseq: An R package for identifying differentially expressed genes from RNA-seq data","volume":"26","author":"Wang","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/gigascience\/gix087","article-title":"Lipidomics profiling reveals the role of glycerophospholipid metabolism in psoriasis","volume":"6","author":"Zeng","year":"2017","journal-title":"Gigascience"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8532\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:38:09Z","timestamp":1760035089000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8532"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,2]]},"references-count":33,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["ijms26178532"],"URL":"https:\/\/doi.org\/10.3390\/ijms26178532","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,2]]}}}