{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T21:20:32Z","timestamp":1790198432424,"version":"4.1.0"},"reference-count":37,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2025,6,25]],"date-time":"2025-06-25T00:00:00Z","timestamp":1750809600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Bolyai J\u00e1nos postdoctoral research fellow fund","award":["GINOP_PLUSZ-2.1.1-21-2022-00083"],"award-info":[{"award-number":["GINOP_PLUSZ-2.1.1-21-2022-00083"]}]},{"name":"Bolyai J\u00e1nos postdoctoral research fellow fund","award":["SZTE_SZAOK-KKA_SZAOK-SZBK-2023\/5S809"],"award-info":[{"award-number":["SZTE_SZAOK-KKA_SZAOK-SZBK-2023\/5S809"]}]},{"name":"Incubation Competence Centre of the Centre of Excellence for Interdisciplinary Research, Development, and Innovation of the University of Szeged","award":["GINOP_PLUSZ-2.1.1-21-2022-00083"],"award-info":[{"award-number":["GINOP_PLUSZ-2.1.1-21-2022-00083"]}]},{"name":"Incubation Competence Centre of the Centre of Excellence for Interdisciplinary Research, Development, and Innovation of the University of Szeged","award":["SZTE_SZAOK-KKA_SZAOK-SZBK-2023\/5S809"],"award-info":[{"award-number":["SZTE_SZAOK-KKA_SZAOK-SZBK-2023\/5S809"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Peptidylarginine deiminase 4 (PAD4) catalyzes protein citrullination, a post-translational modification implicated in type 1 diabetes mellitus (T1DM). This study examined PAD4 expression and activity in the pancreas of streptozotocin (STZ)-induced diabetic Wistar rats. Animals were divided into three groups: (A) STZ-induced diabetic rats (60 mg\/kg, i.p.), (B) non-diabetic controls, and (C) diabetic rats treated with Cl-amidine (5 mg\/kg), a pan-PAD inhibitor, from week six post-induction. Analyses included PAD4 mRNA and protein expression, citrullinated histone H3 (CitH3), calcium concentration, and neutrophil elastase activity. Diabetic rats exhibited increased PAD4 expression, CitH3 levels, and NETosis markers, alongside reduced pancreatic calcium, suggesting calcium consumption during PAD4 activation. Cl-amidine treatment attenuated NETosis. These results implicate PAD4 in T1DM pathogenesis via NETosis and support the utility of STZ-induced diabetic rats as a model for PAD4-targeted studies. Cl-amidine may represent a promising therapeutic approach to reduce pancreatic inflammation in T1DM.<\/jats:p>","DOI":"10.3390\/ijms26136113","type":"journal-article","created":{"date-parts":[[2025,6,26]],"date-time":"2025-06-26T06:55:13Z","timestamp":1750920913000},"page":"6113","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Targeting PAD4: A Promising Strategy to Combat \u03b2-Cell Loss in Type 1 Diabetes"],"prefix":"10.3390","volume":"26","author":[{"given":"Hsu Lin","family":"Kang","sequence":"first","affiliation":[{"name":"Department of Oral Biology and Experimental Dental Research, Faculty of Dentistry, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e1s","family":"Sz\u00e1sz","sequence":"additional","affiliation":[{"name":"Department of Oral Biology and Experimental Dental Research, Faculty of Dentistry, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsuzsanna","family":"Valkusz","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Albert Szent-Gy\u00f6rgyi Medical School, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2057-7993","authenticated-orcid":false,"given":"Tam\u00e1s","family":"V\u00e1rkonyi","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Albert Szent-Gy\u00f6rgyi Medical School, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anik\u00f3","family":"P\u00f3sa","sequence":"additional","affiliation":[{"name":"Department of Oral Biology and Experimental Dental Research, Faculty of Dentistry, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krisztina","family":"Kupai","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Albert Szent-Gy\u00f6rgyi Medical School, University of Szeged, 6703 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1662","DOI":"10.1016\/j.biocel.2006.03.008","article-title":"Citrullination: A Posttranslational Modification in Health and Disease","volume":"38","author":"Tarcsa","year":"2006","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"baab012","DOI":"10.1093\/database\/baab012","article-title":"Post-Translational Modifications in Proteins: Resources, Tools and Prediction Methods","volume":"2021","author":"Ramazi","year":"2021","journal-title":"Database"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s11892-016-0752-4","article-title":"Where, How, and When: Positioning Posttranslational Modification Within Type 1 Diabetes Pathogenesis","volume":"16","author":"McLaughlin","year":"2016","journal-title":"Curr. Diab. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1111\/jth.13951","article-title":"Citrullinated Histone H3, a Biomarker of Neutrophil Extracellular Trap Formation, Predicts the Risk of Venous Thromboembolism in Cancer Patients","volume":"16","author":"Mauracher","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e2310320","DOI":"10.1002\/adma.202310320","article-title":"Neutrophil Function Conversion Driven by Immune Switchpoint Regulator against Diabetes-Related Biofilm Infections","volume":"36","author":"Guo","year":"2024","journal-title":"Adv. Mater."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1038\/nm.3887","article-title":"Diabetes Primes Neutrophils to Undergo NETosis, Which Impairs Wound Healing","volume":"21","author":"Wong","year":"2015","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Yang, M.-L., Sodr\u00e9, F.M.C., Mamula, M.J., and Overbergh, L. (2021). Citrullination and PAD Enzyme Biology in Type 1 Diabetes\u2014Regulators of Inflammation, Autoimmunity, and Pathology. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.678953"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"174623","DOI":"10.1016\/j.ejphar.2021.174623","article-title":"Inhibition of PAD4-Mediated NET Formation by Cl-Amidine Prevents Diabetes Development in Nonobese Diabetic Mice","volume":"916","author":"Shen","year":"2022","journal-title":"Eur. J. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"113840","DOI":"10.1016\/j.ejmech.2021.113840","article-title":"Peptidylarginine Deiminases 4 as a Promising Target in Drug Discovery","volume":"226","author":"Yang","year":"2021","journal-title":"Eur. J. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"He, W., Xi, Q., Cui, H., Zhang, P., Huang, R., Wang, T., and Wang, D. (2022). Forsythiaside B Ameliorates Coagulopathies in a Rat Model of Sepsis through Inhibition of the Formation of PAD4-Dependent Neutrophil Extracellular Traps. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.1022985"},{"key":"ref_11","unstructured":"(2025, March 22). Streptozotocin-Induced Diabetic Models in Mice and Rats-Furman-2021-Current Protocols-Wiley Online Library. Available online: https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full\/10.1002\/cpz1.78."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zhu, Y., Xia, X., He, Q., Xiao, Q.-A., Wang, D., Huang, M., and Zhang, X. (2023). Diabetes-Associated Neutrophil NETosis: Pathogenesis and Interventional Target of Diabetic Complications. Front. Endocrinol., 14.","DOI":"10.3389\/fendo.2023.1202463"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"109134","DOI":"10.1016\/j.clim.2022.109134","article-title":"Citrullination: A Modification Important in the Pathogenesis of Autoimmune Diseases","volume":"245","author":"Zhu","year":"2022","journal-title":"Clin. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/j.autrev.2016.02.018","article-title":"A Novel Method for High-Throughput Detection and Quantification of Neutrophil Extracellular Traps Reveals ROS-Independent NET Release with Immune Complexes","volume":"15","author":"Kraaij","year":"2016","journal-title":"Autoimmun. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1002\/path.4359","article-title":"Neutrophil Extracellular Traps Promote Differentiation and Function of Fibroblasts","volume":"233","author":"Chrysanthopoulou","year":"2014","journal-title":"J. Pathol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e37015","DOI":"10.1097\/MD.0000000000037015","article-title":"The Investigation of Cytotoxic and Apoptotic Activity of Cl-Amidine on the Human U-87 MG Glioma Cell Line","volume":"103","author":"Dikmen","year":"2024","journal-title":"Medicine"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Yao, H., Cao, G., Liu, Z., Zhao, Y., Yan, Z., Wang, S., Wang, Y., Guo, Z., and Wang, Y. (2022). Inhibition of Netosis with PAD Inhibitor Attenuates Endotoxin Shock Induced Systemic Inflammation. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113264"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s13577-022-00822-1","article-title":"Cl-Amidine Attenuates Lipopolysaccharide-Induced Inflammation in Human Gingival Fibroblasts via the JNK\/MAPK, NF-\u03baB, and Nrf2 Signalling Pathways","volume":"36","author":"Du","year":"2023","journal-title":"Hum. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1039\/D3BM01904G","article-title":"Efficient Healing of Diabetic Wounds by MSC-EV-7A Composite Hydrogel via Suppression of Inflammation and Enhancement of Angiogenesis","volume":"12","author":"Long","year":"2024","journal-title":"Biomater. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"347","DOI":"10.7150\/ijbs.78046","article-title":"Neutrophil Extracellular Traps Delay Diabetic Wound Healing by Inducing Endothelial-to-Mesenchymal Transition via the Hippo Pathway","volume":"19","author":"Yang","year":"2023","journal-title":"Int. J. Biol. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Altamura, S., Lombardi, F., Palumbo, P., Cinque, B., Ferri, C., Del Pinto, R., and Pietropaoli, D. (2024). The Evolving Role of Neutrophils and Neutrophil Extracellular Traps (NETs) in Obesity and Related Diseases: Recent Insights and Advances. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252413633"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6963","DOI":"10.1007\/s11033-023-08535-w","article-title":"The Role of GLUT2 in Glucose Metabolism in Multiple Organs and Tissues","volume":"50","author":"Sun","year":"2023","journal-title":"Mol. Biol. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1021\/acs.accounts.9b00024","article-title":"Protein Arginine Deiminases (PADs): Biochemistry and Chemical Biology of Protein Citrullination","volume":"52","author":"Mondal","year":"2019","journal-title":"Acc. Chem. Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Zhou, Y., Chen, B., Mittereder, N., Chaerkady, R., Strain, M., An, L.-L., Rahman, S., Ma, W., Low, C.P., and Chan, D. (2017). Spontaneous Secretion of the Citrullination Enzyme PAD2 and Cell Surface Exposure of PAD4 by Neutrophils. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.01200"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Sharma, C., Hamza, A., Boyle, E., Donu, D., and Cen, Y. (2024). Post-Translational Modifications and Diabetes. Biomolecules, 14.","DOI":"10.3390\/biom14030310"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1097\/MED.0000000000000485","article-title":"Neutrophils and Type 1 Diabetes: Current Knowledge and Suggested Future Directions","volume":"26","author":"Battaglia","year":"2019","journal-title":"Curr. Opin. Endocrinol. Diabetes Obes."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1007\/s00018-022-04126-3","article-title":"Citrullination in the Pathology of Inflammatory and Autoimmune Disorders: Recent Advances and Future Perspectives","volume":"79","author":"Ciesielski","year":"2022","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Aukrust, S.G., Holte, K.B., Opstad, T.B., Seljeflot, I., Berg, T.J., and Helseth, R. (2022). NETosis in Long-Term Type 1 Diabetes Mellitus and Its Link to Coronary Artery Disease. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.799539"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.ceca.2011.08.005","article-title":"The Central Role of Calcium in the Effects of Cytokines on Beta-Cell Function: Implications for Type 1 and Type 2 Diabetes","volume":"50","author":"Ramadan","year":"2011","journal-title":"Cell Calcium"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"182","DOI":"10.14712\/fb2017063050182","article-title":"Original Article Cytokine Network & NETs","volume":"63","author":"Garley","year":"2017","journal-title":"Folia Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/bj20030407","article-title":"Principles of Interleukin (IL)-6-Type Cytokine Signalling and Its Regulation","volume":"374","author":"Heinrich","year":"2003","journal-title":"Biochem. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"920613","DOI":"10.1155\/2014\/920613","article-title":"Cytokines and Diabetes Research","volume":"2014","author":"Xiao","year":"2014","journal-title":"J. Diabetes Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1046\/j.1365-3083.2002.01046.x","article-title":"The Effect of LPS on Neutrophils from Patients with High Risk of Type 1 Diabetes Mellitus in Relation to IL-8, IL-10 and IL-12 Production and Apoptosis In Vitro: Polymorphonuclear Neutrophils Apoptosis in DM1","volume":"55","author":"Glowacka","year":"2002","journal-title":"Scand. J. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"516","DOI":"10.2337\/db20-0421","article-title":"Peptidylarginine Deiminase Inhibition Prevents Diabetes Development in NOD Mice","volume":"70","author":"Bissenova","year":"2021","journal-title":"Diabetes"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Abacar, K., Macleod, T., Direskeneli, H., and McGonagle, D. (2024). How Underappreciated Autoinflammatory (Innate Immunity) Mechanisms Dominate Disparate Autoimmune Disorders. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1439371"},{"key":"ref_36","first-page":"274","article-title":"Streptozotocin as a Tool for Induction of Rat Models of Diabetes: A Practical Guide","volume":"22","author":"Ghasemi","year":"2023","journal-title":"Excli J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"101331","DOI":"10.1016\/j.trim.2020.101331","article-title":"Sex-Dependent Mechanisms Involved in Renal Tolerance to Ischemia-Reperfusion: Role of Inflammation and Histone H3 Citrullination","volume":"63","author":"Melo","year":"2020","journal-title":"Transpl. Immunol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6113\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:58:37Z","timestamp":1760032717000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/13\/6113"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,25]]},"references-count":37,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["ijms26136113"],"URL":"https:\/\/doi.org\/10.3390\/ijms26136113","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,25]]}}}