{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T02:20:07Z","timestamp":1788402007336,"version":"build-2803163510"},"reference-count":46,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T00:00:00Z","timestamp":1758844800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007065","name":"University of Salerno","doi-asserted-by":"publisher","award":["ORSA243414"],"award-info":[{"award-number":["ORSA243414"]}],"id":[{"id":"10.13039\/501100007065","id-type":"DOI","asserted-by":"publisher"}]},{"name":"\u201cTHE \u2013 Tuscany Health Ecosystem\u201d Italian Ministry of University and Research","award":["B83C22003920001"],"award-info":[{"award-number":["B83C22003920001"]}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["IJMS"],"abstract":"<jats:p>Oxidative stress and mitochondrial dysfunction play a key role in the early stage of Doxorubicin (Doxo)-induced cardiotoxicity. Our study investigated the potential cardioprotective role of Simvastatin (Sim), widely known for its antioxidant properties, in an in vitro model of Doxo-induced acute cardiotoxicity. Human Cardiomyocytes (HCMs) were treated with Sim (10 \u00b5M, 4 h) and then co-exposed to Doxo (1 \u00b5M) and Sim for 20 h. Our data showed that Sim co-treatment significantly (p &lt; 0.05) reduced both cytosolic and mitochondrial Doxo-induced reactive oxygen species overproduction. In Sim co-treated cells, significant reductions in nuclear factor erythroid 2-related factor 2 (Nrf2) gene expression (p &lt; 0.01) and catalase (CAT), heme-oxygenase 1 (HO-1), and superoxide dismutase 2 (SOD2) levels (p &lt; 0.05) compared to Doxo-treated cells were also demonstrated, suggesting a decreased need for compensatory antioxidant defense responses. Moreover, significant reductions in Doxo-induced mitochondrial calcium overload, mitochondrial membrane depolarization (p &lt; 0.005), and apoptosis (p &lt; 0.005) confirmed the protective effects of Sim co-treatment on cardiomyocytes. These data confirm that Sim could be a valuable therapeutic strategy for reducing Doxo-induced HCM damage, preventing the development of dilated cardiomyopathy and long-term heart damage, which are the main limitations of anthracycline use. Finally, real-time PCR analysis revealed that Sim co-treatment significantly reduced (p &lt; 0.001) the Doxo-induced overexpression of MAP4K4, a mitogen-activated protein kinase kinase kinase kinase-4 (MAP4K4) involved in oxidative stress-induced cell death, thus suggesting the involvement of other molecular mechanisms in Sim-mediated cardioprotection.<\/jats:p>","DOI":"10.3390\/ijms26199440","type":"journal-article","created":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T15:25:59Z","timestamp":1758900359000},"page":"9440","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Cardioprotective Effects of Simvastatin in Doxorubicin-Induced Acute Cardiomyocyte Injury"],"prefix":"10.3390","volume":"26","author":[{"given":"Roberta","family":"Vitale","sequence":"first","affiliation":[{"name":"Department of Pharmacy, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariangela","family":"Mazzone","sequence":"additional","affiliation":[{"name":"Department of Innovative Technologies in Medicine and Dentistry, University \u201cG. d\u2019Annunzio\u201d Chieti-Pescara, 66100 Chieti, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9308-9857","authenticated-orcid":false,"given":"Maria Carmela","family":"Di Marcantonio","sequence":"additional","affiliation":[{"name":"Department of Innovative Technologies in Medicine and Dentistry, University \u201cG. d\u2019Annunzio\u201d Chieti-Pescara, 66100 Chieti, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2333-1630","authenticated-orcid":false,"given":"Stefania","family":"Marzocco","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7127-0938","authenticated-orcid":false,"given":"Gabriella","family":"Mincione","sequence":"additional","affiliation":[{"name":"Department of Innovative Technologies in Medicine and Dentistry, University \u201cG. d\u2019Annunzio\u201d Chieti-Pescara, 66100 Chieti, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9843-8443","authenticated-orcid":false,"given":"Ada","family":"Popolo","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Martin, J., Ells, Z., Dolle, B., Rouzina, I.F., Williams, M.C., and Paramanathan, T. (2024). Optical tweezers reveal nanomolar DNA binding affinity of doxorubicin. Biophys. J., 123.","DOI":"10.1016\/j.bpj.2023.11.1429"},{"key":"ref_2","first-page":"363","article-title":"Cancer treatment and survivorship statistics, 2019","volume":"69","author":"Miller","year":"2019","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1038\/s41419-021-03614-x","article-title":"Regulated cell death pathways in doxorubicin-induced cardiotoxicity","volume":"12","author":"Christidi","year":"2021","journal-title":"Cell. Death Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1007\/s10741-024-10414-w","article-title":"The significance of the apelinergic system in doxorubicin-induced cardiotoxicity","volume":"29","author":"Matusik","year":"2024","journal-title":"Heart Fail. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Tranchita, E., Murri, A., Grazioli, E., Cerulli, C., Emerenziani, G.P., Ceci, R., Caporossi, D., Dimauro, I., and Parisi, A. (2022). The Beneficial Role of Physical Exercise on Anthracyclines Induced Cardiotoxicity in Breast Cancer Patients. Cancers, 14.","DOI":"10.3390\/cancers14092288"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Sangweni, N.F., van Vuuren, D., Mabasa, L., Gabuza, K., Huisamen, B., Naidoo, S., Barry, R., and Johnson, R. (2022). Prevention of Anthracycline-Induced Cardiotoxicity: The Good and Bad of Current and Alternative Therapies. Front. Cardiovasc. Med., 9.","DOI":"10.3389\/fcvm.2022.907266"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Rawat, P.S., Jaiswal, A., Khurana, A., Bhatti, J.S., and Navik, U. (2021). Doxorubicin-induced cardiotoxicity: An update on the molecular mechanism and novel therapeutic strategies for effective management. Biomed. Pharmacother., 139.","DOI":"10.1016\/j.biopha.2021.111708"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Pecoraro, M., Marzocco, S., Belvedere, R., Petrella, A., Franceschelli, S., and Popolo, A. (2023). Simvastatin Reduces Doxorubicin-Induced Cardiotoxicity: Effects beyond Its Antioxidant Activity. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24087573"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107683","DOI":"10.1016\/j.carpath.2024.107683","article-title":"Doxorubicin-related cardiotoxicity: Review of fundamental pathways of cardiovascular system injury","volume":"73","author":"Avagimyan","year":"2024","journal-title":"Cardiovasc. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1521020","DOI":"10.1155\/2017\/1521020","article-title":"Oxidative stress and cellular response to doxorubicin: A common factor in the complex milieu of anthracycline cardiotoxicity","volume":"2017","author":"Cappetta","year":"2017","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1016\/j.jacbts.2023.10.009","article-title":"Molecular Mechanisms and Therapeutic Targeting of Ferroptosis in Doxorubicin-Induced Cardiotoxicity","volume":"9","author":"Wu","year":"2024","journal-title":"JACC Basic Transl. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Sadowska, A., Osi\u0144ski, P., Roztocka, A., Kaczmarz-Chojnacka, K., Zapora, E., Sawicka, D., and Car, H. (2024). Statins\u2014From Fungi to Pharmacy. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25010466"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.26420\/austincardiol.2023.1037","article-title":"Statins to prevent anthracyclines-induced cardiotoxicity","volume":"8","author":"Ma","year":"2023","journal-title":"Austin Cardio"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1429","DOI":"10.1007\/s12012-025-10030-6","article-title":"Statins: Novel Approaches for the Management of Doxorubicin-Induced Cardiotoxicity-A Literature Review","volume":"25","author":"TamehriZadeh","year":"2025","journal-title":"Cardiovasc. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"102130","DOI":"10.1016\/j.cpcardiol.2023.102130","article-title":"Role of Statin Therapy in Prevention of Anthracycline-Induced Cardiotoxicity: A Three Dimentional Echocardiography Study","volume":"49","author":"Mohamed","year":"2024","journal-title":"Curr. Probl. Cardiol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jiang, R., Lou, L., Shi, W., Chen, Y., Fu, Z., Liu, S., Sok, T., Li, Z., Zhang, X., and Yang, J. (2024). Statins in Mitigating Anticancer Treatment-Related Cardiovascular Disease. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms251810177"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Vitale, R., Marzocco, S., and Popolo, A. (2024). Role of Oxidative Stress and Inflammation in Doxorubicin-Induced Cardiotoxicity: A Brief Account. Int. J. Mol. Sci., 25.","DOI":"10.20944\/preprints202405.1681.v1"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.toxlet.2019.02.013","article-title":"Oxidative stress injury in doxorubicin-induced cardiotoxicity","volume":"307","author":"Songbo","year":"2019","journal-title":"Toxicol. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2848","DOI":"10.1007\/s12011-021-02891-z","article-title":"Selenium Attenuates Doxorubicin-Induced Cardiotoxicity Through Nrf2-NLRP3 Pathway","volume":"200","author":"Yang","year":"2022","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Bkaily, G., and Danielle, J. (2023). Calcium Homeostasis, Transporters, and Blockers in Health and Diseases of the Cardiovascular System. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24108803"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1007\/s12012-014-9305-8","article-title":"Doxorubicin-Mediated Cardiotoxicity: Role of Mitochondrial Connexin 43","volume":"15","author":"Pecoraro","year":"2015","journal-title":"Cardiovasc. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"795602","DOI":"10.1155\/2015\/795602","article-title":"Chemotherapy-Induced Cardiotoxicity: Overview of the Roles of Oxidative Stress","volume":"2015","author":"Angsutararux","year":"2015","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1016\/j.cstres.2024.09.001","article-title":"The mechanism and therapeutic strategies in doxorubicin induced cardiotoxicity: Role of programmed cell death","volume":"29","author":"Li","year":"2024","journal-title":"Cell Stress Chaperones"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1016\/j.stem.2020.01.015","article-title":"MAP4K4 Inhibition Promotes Survival of Human Stem Cell-Derived Cardiomyocytes and Reduces Infarct Size In Vivo","volume":"26","author":"Fiedler","year":"2020","journal-title":"Cell Stem Cell."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Golforoush, P.A., Narasimhan, P., Chaves-Guerrero, P.P., Lawrence, E., Newton, G., Yan, R., Harding, S.E., Perrior, T., Chapman, K.L., and Schneider, M.D. (2020). Selective protection of human cardiomyocytes from anthracycline cardiotoxicity by small molecule inhibitors of MAP4K4. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-68907-1"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Al Khafaji, A.T., Barakat, A.M., Shayyal, A.J., Taan, A.A., and Aboqader Al-Aouadi, R.F. (2025). Managing Doxorubicin Cardiotoxicity: Insights Into Molecular Mechanisms and Protective Strategies. J. Biochem. Mol. Toxicol., 39.","DOI":"10.1002\/jbt.70155"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Kundnani, N.R., Passini, V., Stefania Carlogea, I., Dumitrescu, P., Meche, V., Buzas, R., and Duda-Seiman, D.M. (2025). Overview of Oncology: Drug-Induced Cardiac Toxicity. Medicina, 61.","DOI":"10.3390\/medicina61040709"},{"key":"ref_28","first-page":"2193","article-title":"Simvastatin protects cardiomyocytes from doxorubicin cardiotoxicity by suppressing endoplasmic reticulum stress and activating Akt signaling","volume":"9","author":"Liu","year":"2016","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Vitale, R., Marzocco, S., and Popolo, A. (2025). Simvastatin Enhances the Cytotoxic Effects of Doxorubicin in a Mammary Adenocarcinoma Cell Model by Involving Connexin 43. J. Biochem. Mol. Toxicol., 39.","DOI":"10.1002\/jbt.70214"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Quagliariello, V., Berretta, M., Bisceglia, I., Giacobbe, I., Iovine, M., Giordano, V., Arianna, R., Barbato, M., Izzo, F., and Maurea, C. (2024). The sGCa Vericiguat Exhibit Cardioprotective and Anti-Sarcopenic Effects through NLRP-3 Pathways: Potential Benefits for Anthracycline-Treated Cancer Patients. Cancers, 16.","DOI":"10.3390\/cancers16081487"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1186\/s12935-023-02890-1","article-title":"Paradoxical effects of statins on endothelial and cancer cells: The impact of concentrations","volume":"23","author":"Ahmadi","year":"2023","journal-title":"Cancer Cell Int."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1007\/s11912-024-01579-6","article-title":"Statins for the Primary Prevention of Anthracycline Cardiotoxicity: A Comprehensive Review","volume":"26","author":"Bhasin","year":"2024","journal-title":"Curr. Oncol. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/S0167-5273(02)00577-6","article-title":"A comparative study of the in vitro antioxidant activity of statins","volume":"90","author":"Franzoni","year":"2003","journal-title":"Int. J. Cardiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"8863789","DOI":"10.1155\/2021\/8863789","article-title":"Role of Oxidative Stress in the Mechanisms of Anthracycline-Induced Cardiotoxicity: Effects of Preventive Strategies","volume":"2021","author":"Carrasco","year":"2021","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1007\/s10753-023-01908-0","article-title":"The Role of Nrf2 and Inflammation on the Dissimilar Cardiotoxicity of Doxorubicin in Two-Time Points: A Cardio-Oncology In Vivo Study Through Time","volume":"47","author":"Ferreira","year":"2024","journal-title":"Inflammation"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/s12964-023-01077-5","article-title":"Role of oxidative stress and inflammation-related signaling pathways in doxorubicin-induced cardiomyopathy","volume":"21","author":"Shi","year":"2023","journal-title":"Cell Commun. Signal."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2672","DOI":"10.1073\/pnas.1821022116","article-title":"Ferroptosis as a target for protection against cardiomyopathy","volume":"116","author":"Fang","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2713","DOI":"10.1007\/s12010-024-05141-9","article-title":"Metformin Alleviates Doxorubicin-Induced Cardiotoxicity via Preserving Mitochondrial Dynamics Balance and Calcium Homeostasis","volume":"197","author":"Maghraby","year":"2025","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.bbrc.2015.01.137","article-title":"Calcium and mitochondria in the regulation of cell death","volume":"460","author":"Orrenius","year":"2015","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Sripusanapan, A., Piriyakulthorn, C., Apaijai, N., Chattipakorn, S.C., and Chattipakorn, N. (2025). Ivabradine ameliorates doxorubicin-induced cardiotoxicity through improving mitochondrial function and cardiac calcium homeostasis. Biochem. Pharmacol., 236.","DOI":"10.1016\/j.bcp.2025.116881"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1177\/0960327111429141","article-title":"Statins lower calcium-induced oxidative stress in isolated mitochondria","volume":"31","author":"Parihar","year":"2011","journal-title":"Hum. Exp. Toxicol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Gao, X., Gao, C., Liu, G., and Hu, J. (2016). MAP4K4: An emerging therapeutic target in cancer. Cell Biosci., 6.","DOI":"10.1186\/s13578-016-0121-7"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"111213","DOI":"10.1016\/j.cellsig.2024.111213","article-title":"Regulatory effects of statins on Akt signaling for prevention of cancers","volume":"120","author":"Hosseini","year":"2024","journal-title":"Cell Signal."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1111\/fcp.12981","article-title":"Repurposing of atorvastatin and metformin denotes their individual and combined antiproliferative effects in non-small cell lung cancer","volume":"38","author":"Salim","year":"2024","journal-title":"Fundam. Clin. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1139\/Y10-100","article-title":"Cytotoxic activity of nemorosone in human MCF-7 breast cancer cells","volume":"89","author":"Popolo","year":"2011","journal-title":"Can. J. Physiol. Pharmacol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Pecoraro, M., Marzocco, S., Franceschelli, S., and Popolo, A. (2022). Trastuzumab and Doxorubicin Sequential Administration Increases Oxidative Stress and Phosphorylation of Connexin 43 on Ser368. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23126375"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/19\/9440\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,29]],"date-time":"2025-09-29T04:26:28Z","timestamp":1759119988000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/19\/9440"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,26]]},"references-count":46,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["ijms26199440"],"URL":"https:\/\/doi.org\/10.3390\/ijms26199440","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,26]]}}}