{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T05:21:24Z","timestamp":1766726484907,"version":"3.48.0"},"reference-count":45,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T00:00:00Z","timestamp":1766534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>We explored the ability of the low molecular weight, polyphenol-rich fractions obtained from chestnut shells to inhibit ferroptosis in Friedreich Ataxia (FRDA), an inherited neuro- and cardio-degenerative disease. We prepared an aqueous extract by an eco-sustainable method and obtained a polyphenol-rich fraction (fraction D) of molecular weight less than 1.0 kDa after molecular size fractionation. The total phenols were 173.28 \u00b1 4.97 \u03bcg gallic acid equivalents\/mg fraction, and analysis by UHPLC-ITMSn and RP-HPLC-UV revealed thirteen phenolic compounds with gallic acid and protocatechuic acid (PCA) as the most abundant (26.29 \u00b1 2.19 and 4.93 \u00b1 0.19 \u03bcg\/mg fraction, respectively). Using a cellular assay based on patient-derived FRDA fibroblasts, we observed that chestnut shell dry extract at 20 \u00b5g\/mL increased the survival of cells stressed with the ferroptosis inducer erastin from 8% to 45% and that this activity was dose-dependent. Fraction D at 20 \u00b5g\/mL showed similar strong activity, increasing cell survival from 0.5% to 14% and decreasing lipid peroxidation by 42%. PCA, the most efficacious compound, doubled cell survival and decreased lipid peroxidation by 20%. Moreover, PCA increased the survival of cells in which frataxin was knocked down 1.5-fold and decreased ALOX12 expression. Our data suggest that PCA could be a promising molecule to explore FRDA pathophysiology.<\/jats:p>","DOI":"10.3390\/molecules31010070","type":"journal-article","created":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T12:22:27Z","timestamp":1766578947000},"page":"70","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Polyphenol-Enriched Fraction from Chestnut Shells as a Source of Bioactive Compounds for Friedreich Ataxia"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9310-3564","authenticated-orcid":false,"given":"Giuseppe","family":"Squillaci","sequence":"first","affiliation":[{"name":"Institute of Biochemistry and Cell Biology, National Research Council of Italy (CNR), Via Pietro Castellino 111, 80131 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Grazia M.","family":"Cotticelli","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, 3501 Civic Center Blvd., Philadelphia, PA 19104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0214-3905","authenticated-orcid":false,"given":"Virginia","family":"Carbone","sequence":"additional","affiliation":[{"name":"Institute of Food Sciences, National Research Council of Italy (CNR), Via Roma 64, 83100 Avellino, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Avery O.","family":"Westfall","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, 3501 Civic Center Blvd., Philadelphia, PA 19104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6059-9762","authenticated-orcid":false,"given":"Robert B.","family":"Wilson","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Children\u2019s Hospital of Philadelphia, 3501 Civic Center Blvd., Philadelphia, PA 19104, USA"},{"name":"Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 3400 Spruce St., Philadelphia, PA 19104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3189-2526","authenticated-orcid":false,"given":"Alessandra","family":"Morana","sequence":"additional","affiliation":[{"name":"Research Institute on Terrestrial Ecosystems, National Research Council of Italy (CNR), via Pietro Castellino 111, 80131 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1016\/S1474-4422(25)00175-9","article-title":"FACROSS study group. Friedreich\u2019s ataxia-a rare multisystem disease","volume":"24","author":"Reetz","year":"2025","journal-title":"Lancet Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1038\/s41582-025-01065-y","article-title":"A global perspective on research advances and future challenges in Friedreich Ataxia","volume":"21","author":"Indelicato","year":"2025","journal-title":"Nat. Rev. Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"NS20200093","DOI":"10.1042\/NS20200093","article-title":"Mitochondrial and metabolic dysfunction in Friedreich Ataxia: Update on pathophysiological relevance and clinical interventions","volume":"5","author":"Lynch","year":"2021","journal-title":"Neuronal Signal."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2152","DOI":"10.1096\/fj.09-143222","article-title":"Does oxidative stress contribute to the pathology of Friedreich\u2019s ataxia? A radical question","volume":"24","author":"Armstrong","year":"2010","journal-title":"FASEB J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.redox.2013.06.004","article-title":"Insights into the role of oxidative stress in the pathology of Friedreich Ataxia using peroxidation resistant polyunsaturated fatty acids","volume":"1","author":"Cotticelli","year":"2013","journal-title":"Redox Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Turchi, R., Faraonio, R., Lettieri-Barbato, D., and Aquilano, K. (2020). An Overview of the Ferroptosis Hallmarks in Friedreich\u2019s Ataxia. Biomolecules, 10.","DOI":"10.3390\/biom10111489"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1016\/j.cell.2012.03.042","article-title":"Ferroptosis: An iron-dependent form of nonapoptotic cell death","volume":"149","author":"Dixon","year":"2012","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"E4966","DOI":"10.1073\/pnas.1603244113","article-title":"Peroxidation of polyunsaturated fatty acids by lipoxygenases drives ferroptosis","volume":"113","author":"Yang","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1021\/acscentsci.7b00589","article-title":"Resolving the Role of Lipoxygenases in the Initiation and Execution of Ferroptosis","volume":"4","author":"Shah","year":"2018","journal-title":"ACS Cent. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.chembiol.2008.02.010","article-title":"Synthetic lethal screening identifies compounds activating iron-dependent, nonapoptotic cell death in oncogenic-RAS-harboring cancer cells","volume":"15","author":"Yang","year":"2008","journal-title":"Chem. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Edzeamey, F.J., Ramchunder, Z., Pourzand, C., and Anjomani Virmouni, S. (2024). Emerging antioxidant therapies in Friedreich\u2019s ataxia. Front. Pharmacol., 15.","DOI":"10.3389\/fphar.2024.1359618"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1002\/acn3.660","article-title":"Safety, pharmacodynamics, and potential benefit of omaveloxolone in Friedreich Ataxia","volume":"6","author":"Lynch","year":"2019","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/s41417-020-0170-2","article-title":"Transcription factors in ferroptotic cell death","volume":"27","author":"Dai","year":"2020","journal-title":"Cancer Gene Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"100377","DOI":"10.1016\/j.prmcm.2024.100377","article-title":"Natural flavonoids act as potent ferroptosis inhibitors and their potentials in the treatment of ferroptosis-associated diseases","volume":"10","author":"Liu","year":"2024","journal-title":"Pharmacol. Res.-Mod. Chin. Med."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Lesjak, M., Simin, N., and Srai, S.K.S. (2022). Can Polyphenols Inhibit Ferroptosis?. Antioxidants, 11.","DOI":"10.3390\/antiox11010150"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e14264","DOI":"10.1111\/jfbc.14264","article-title":"Health benefits of polyphenols: A concise review","volume":"46","author":"Rana","year":"2022","journal-title":"J. Food Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.egypro.2017.07.269","article-title":"AgroCycle\u2014Developing a circular economy in agriculture","volume":"123","author":"Toop","year":"2017","journal-title":"Energy Procedia"},{"key":"ref_18","unstructured":"Singh, S., Singh, P., Sharma, A., and Choudhury, M. (2023). Agricultural Waste as a Resource: The Lesser Travelled Road to Sustainability. Agriculture Waste Management and Bioresource: The Circular Economy Perspective, John Wiley & Sons Ltd.. [1st ed.]."},{"key":"ref_19","first-page":"246","article-title":"Chestnut tannin: New use, research and bioeconomy","volume":"9","author":"Ciriminna","year":"2024","journal-title":"J. Bioresour. Bioprod."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.cej.2012.06.122","article-title":"Optimization of antioxidants\u2014Extraction from Castanea sativa leaves","volume":"203","author":"Couto","year":"2012","journal-title":"Chem. Eng. J."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Cerulli, A., Napolitano, A., Hosek, J., Masullo, M., Pizza, C., and Piacente, S. (2021). Antioxidant and In Vitro Preliminary Anti-Inflammatory Activity of Castanea sativa (Italian Cultivar \u201cMarrone di Roccadaspide\u201d PGI) Burs, Leaves, and Chestnuts Extracts and Their Metabolite Profiles by LC-ESI\/LTQOrbitrap\/MS\/MS. Antioxidants, 10.","DOI":"10.3390\/antiox10020278"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1016\/j.procbio.2017.09.017","article-title":"Chestnut (Castanea sativa Mill.) industrial wastes as a valued bioresource for the production of active ingredients","volume":"64","author":"Squillaci","year":"2018","journal-title":"Process Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Cacciola, N.A., Squillaci, G., D\u2019Apolito, M., Petillo, O., Veraldi, F., La Cara, F., Peluso, G., Margarucci, S., and Morana, A. (2019). Castanea sativa Mill. Shells Aqueous Extract Exhibits Anticancer Properties Inducing Cytotoxic and Pro-Apoptotic Effects. Molecules, 24.","DOI":"10.20944\/preprints201908.0006.v1"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Trezza, A., Barletta, R., Geminiani, M., Frusciante, L., Olmastroni, T., Sannio, F., Docquier, J.-D., and Santucci, A. (2024). Chestnut Burrs as Natural Source of Antimicrobial Bioactive Compounds: A Valorization of Agri-Food Waste. Appl. Sci., 14.","DOI":"10.3390\/app14156552"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Orso, G., Solovyev, M.M., Facchiano, S., Tyrikova, E., Sateriale, D., Kashinskaya, E., Pagliarulo, C., Hoseinifar, H.S., Simonov, E., and Varricchio, E. (2021). Chestnut Shell Tannins: Effects on Intestinal Inflammation and Dysbiosis in Zebrafish. Animals, 11.","DOI":"10.3390\/ani11061538"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"101806","DOI":"10.1016\/j.scp.2024.101806","article-title":"Solid-liquid extraction of polyphenols from chestnut shells: A sustainable approach for coagulant synthesis","volume":"42","author":"Tomasi","year":"2024","journal-title":"Sustain. Chem. Pharm."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Tong, Z., He, W., Fan, X., and Guo, A. (2022). Biological Function of Plant Tannin and Its Application in Animal Health. Front. Vet. Sci., 8.","DOI":"10.3389\/fvets.2021.803657"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1093\/hmg\/8.3.425","article-title":"The Friedreich\u2019s ataxia mutation confers cellular sensitivity to oxidant stress which is rescued by chelators of iron and calcium and inhibitors of apoptosis","volume":"8","author":"Wong","year":"1999","journal-title":"Hum. Mol. Genet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3055","DOI":"10.1093\/hmg\/11.24.3055","article-title":"A cellular model for Friedreich Ataxia reveals small-molecule glutathione peroxidase mimetics as novel treatment strategy","volume":"11","author":"Jauslin","year":"2002","journal-title":"Hum. Mol. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1124\/jpet.118.252759","article-title":"Ferroptosis as a Novel Therapeutic Target for Friedreich\u2019s Ataxia","volume":"369","author":"Cotticelli","year":"2019","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2535","DOI":"10.1021\/acschemneuro.0c00323","article-title":"Identification of a Novel Oleic Acid Analog with Protective Effects in Multiple Cellular Models of Friedreich Ataxia","volume":"11","author":"Cotticelli","year":"2020","journal-title":"ACS Chem. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3216","DOI":"10.18632\/aging.204009","article-title":"Trehalose inhibits ferroptosis via NRF2\/HO-1 pathway and promotes functional recovery in mice with spinal cord injury","volume":"14","author":"Gong","year":"2022","journal-title":"Aging"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Li, L., Dai, Y., Ke, D., Liu, J., Chen, P., Wei, D., Wang, T., Teng, Y., Yuan, X., and Zhang, Z. (2023). Ferroptosis: New insight into the mechanisms of diabetic nephropathy and retinopathy. Front. Endocrinol., 14.","DOI":"10.3389\/fendo.2023.1215292"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7815","DOI":"10.1038\/s41598-025-92383-0","article-title":"Gallic acid alleviates ferroptosis by negatively regulating APOC3 and improves nerve function deficit caused by traumatic brain injury","volume":"15","author":"Liu","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Tang, H.M., and Cheung, P.C.K. (2019). Gallic Acid Triggers Iron-Dependent Cell Death with Apoptotic, Ferroptotic, and Necroptotic Features. Toxins, 11.","DOI":"10.3390\/toxins11090492"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"902","DOI":"10.1002\/acn3.486","article-title":"4-Hydroxybenzoic acid restores CoQ(10) biosynthesis in human COQ2 deficiency","volume":"4","author":"Herebian","year":"2017","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1021\/acs.biochem.0c00030","article-title":"Ferroptosis Suppressor Protein 1 (FSP1) and Coenzyme Q(10) Cooperatively Suppress Ferroptosis","volume":"59","author":"Hadian","year":"2020","journal-title":"Biochemistry"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1007\/s10565-024-09953-7","article-title":"Protocatechuic acid relieves ferroptosis in hepatic lipotoxicity and steatosis via regulating NRF2 signaling pathway","volume":"40","author":"Feng","year":"2024","journal-title":"Cell Biol. Toxicol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Yang, X., Sun, X., Zhou, F., Xiao, S., Zhong, L., Hu, S., Zhou, Z., Li, L., and Tan, Y. (2023). Protocatechuic Acid Alleviates Dextran-Sulfate-Sodium-Induced Ulcerative Colitis in Mice via the Regulation of Intestinal Flora and Ferroptosis. Molecules, 28.","DOI":"10.3390\/molecules28093775"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"La Rosa, P., Petrillo, S., Fiorenza, M.T., Bertini, E.S., and Piemonte, F. (2020). Ferroptosis in Friedreich\u2019s Ataxia: A Metal-Induced Neurodegenerative Disease. Biomolecules, 10.","DOI":"10.3390\/biom10111551"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1002\/prot.10579","article-title":"Lipoxygenase interactions with natural flavonoid, quercetin, reveal a complex with protocatechuic acid in its X-ray structure at 2.1 A resolution","volume":"54","author":"Borbulevych","year":"2004","journal-title":"Proteins"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1084\/jem.20082596","article-title":"Cardiac 12\/15 lipoxygenase-induced inflammation is involved in heart failure","volume":"206","author":"Kayama","year":"2009","journal-title":"J. Exp. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e185299","DOI":"10.1172\/jci.insight.185299","article-title":"12-Lipoxygenase inhibition delays onset of autoimmune diabetes in human gene replacement mice","volume":"9","author":"Nargis","year":"2024","journal-title":"JCI Insight"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"144","DOI":"10.5344\/ajev.1965.16.3.144","article-title":"Colorimetry of Total Phenolics with Phosphomolybdic-Phosphotungstic Acid Reagents","volume":"16","author":"Singleton","year":"1965","journal-title":"Am. J. Enol. Vitic."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1021\/ac60147a030","article-title":"Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar","volume":"31","author":"Miller","year":"1959","journal-title":"Anal. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/1\/70\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T05:19:36Z","timestamp":1766726376000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/1\/70"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,24]]},"references-count":45,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["molecules31010070"],"URL":"https:\/\/doi.org\/10.3390\/molecules31010070","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2025,12,24]]}}}