{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T04:45:29Z","timestamp":1776660329570,"version":"3.51.2"},"reference-count":46,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T00:00:00Z","timestamp":1776643200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Physiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Human erythrocytes serve as an ideal model for cellular aging, a process where longevity relies on membrane scaffold integrity. The oxidative deterioration of Band 3, a major integral membrane protein, is a central driver of this senescence. This study investigated whether methyl eugenol (ME) stabilizes Band 3 against age-associated oxidative fragility.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Erythrocytes were challenged with H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      to simulate age-associated oxidative injury. Damage was evaluated via hemolysis assays, SEM, and flow cytometry. Sulfate (SO\n                      <jats:sub>4<\/jats:sub>\n                      <jats:sup>2-<\/jats:sup>\n                      ) uptake kinetics and Western blotting were employed to assess Band 3 anion exchange function and structural stability.\n                      <jats:italic>In silico<\/jats:italic>\n                      docking simulated interactions between ME metabolites and the Band 3 structure. Physiological relevance was validated in a human cohort (n = 81; 20\u201390 years) via regression and stratified analyses of glutathione (GSH) and malondialdehyde (MDA) levels.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      ME exhibited an optimal protective concentration at 2 \u00b5M, effectively preserving biconcave morphology and attenuating hemolysis. Treatment significantly mitigated intracellular oxidative stress and rescued cell viability. Mechanistically, ME suppressed the pathological increase in intracellular Ca\n                      <jats:sup>2+<\/jats:sup>\n                      concentration and inhibited calpain activity. Functionally, ME significantly restored sulfate transport rates. Western blotting confirmed that ME specifically preserved the full-length (100 kDa) and cytoplasmic (43 kDa) domains of Band 3, whereas the 55 kDa transmembrane domain remained largely unaffected. Docking simulations predicted a specific interaction with residue ARG292 within the cytoplasmic domain, suggesting a structural basis for this stabilization. In the donor cohort, ME extended the projected GSH half-life (from 47.14 to 64.14 years) and reduced maximal lipid peroxidation by ~40%.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      ME mitigates oxidative eryptosis by coupling Ca\n                      <jats:sup>2+<\/jats:sup>\n                      \u2013calpain inhibition with site-specific Band 3 stabilization, offering a rationale for using ME to standardize erythrocyte quality and reduce age-associated fragility.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fphys.2026.1796160","type":"journal-article","created":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T04:14:18Z","timestamp":1776658458000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Methyl eugenol attenuates age-associated oxidative fragility by coupling Ca2+-calpain inhibition with Band 3 stabilization in human erythrocytes"],"prefix":"10.3389","volume":"17","author":[{"given":"Jiasi","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Breast Surgery, The Tenth Affiliated Hospital, Southern Medical University (Dongguan People's Hospital)","place":["Dongguan, Guangdong, China"]},{"name":"Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology; 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