{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T15:08:53Z","timestamp":1790780933162,"version":"4.1.0"},"reference-count":42,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2023,10,9]],"date-time":"2023-10-09T00:00:00Z","timestamp":1696809600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82104358"],"award-info":[{"award-number":["82104358"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["H2021423066"],"award-info":[{"award-number":["H2021423066"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21372503D"],"award-info":[{"award-number":["21372503D"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["23372502D"],"award-info":[{"award-number":["23372502D"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003787","name":"Hebei Provincial Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82104358"],"award-info":[{"award-number":["82104358"]}],"id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003787","name":"Hebei Provincial Natural Science Foundation of China","doi-asserted-by":"publisher","award":["H2021423066"],"award-info":[{"award-number":["H2021423066"]}],"id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003787","name":"Hebei Provincial Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21372503D"],"award-info":[{"award-number":["21372503D"]}],"id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003787","name":"Hebei Provincial Natural Science Foundation of China","doi-asserted-by":"publisher","award":["23372502D"],"award-info":[{"award-number":["23372502D"]}],"id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]},{"name":"S&amp;T Program of Hebei","award":["82104358"],"award-info":[{"award-number":["82104358"]}]},{"name":"S&amp;T Program of Hebei","award":["H2021423066"],"award-info":[{"award-number":["H2021423066"]}]},{"name":"S&amp;T Program of Hebei","award":["21372503D"],"award-info":[{"award-number":["21372503D"]}]},{"name":"S&amp;T Program of Hebei","award":["23372502D"],"award-info":[{"award-number":["23372502D"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Osteoporosis (OP) is distinguished by a reduction in bone mass and degradation of bone micro-structure, frequently resulting in fractures. As the geriatric demographic expands, the incidence of affected individuals progressively rises, thereby exerting a significant impact on the quality of life experienced by individuals. The flavonoid compound hesperidin has been subject to investigation regarding its effects on skeletal health, albeit the precise mechanisms through which it operates remain ambiguous. This study utilized network pharmacology to predict the core targets and signaling pathways implicated in the anti-OP properties of hesperidin. Molecular docking and molecular dynamics simulations were employed to confirm the stability of the interaction between hesperidin and the core targets. The effects of hesperidin on osteoblastic cells MC3T3-E1 were assessed using MTT, ELISA, alkaline phosphatase assay, and RT-qPCR techniques. Furthermore, in vivo experiments were conducted to determine the potential protective effects of hesperidin on zebrafish bone formation and oxidative stress response. The results demonstrate that network pharmacology has identified 10 key target points, significantly enriched in the estrogen signaling pathway. Hesperidin exhibits notable promotion of MC3T3-E1 cell proliferation and significantly enhances ALP activity. ELISA measurements indicate an elevation in NO levels and a reduction in IL-6 and TNF-\u03b1. Moreover, RT-qPCR analysis consistently reveals that hesperidin significantly modulates the mRNA levels of ESR1, SRC, AKT1, and NOS3 in MC3T3-E1 cells. Hesperidin promotes osteogenesis and reduces oxidative stress in zebrafish. Additionally, we validate the stable and tight binding of hesperidin with ESR1, SRC, AKT1, and NOS3 through molecular dynamics simulations. In conclusion, our comprehensive analysis provides evidence that hesperidin may exert its effects on alleviating OP through the activation of the estrogen signaling pathway via ESR1. This activation leads to the upregulation of SRC, AKT, and eNOS, resulting in an increase in NO levels. Furthermore, hesperidin promotes osteoblast-mediated bone formation and inhibits pro-inflammatory cytokines, thereby alleviating oxidative stress associated with OP.<\/jats:p>","DOI":"10.3390\/molecules28196987","type":"journal-article","created":{"date-parts":[[2023,10,9]],"date-time":"2023-10-09T07:37:50Z","timestamp":1696837070000},"page":"6987","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":36,"title":["Hesperidin Anti-Osteoporosis by Regulating Estrogen Signaling Pathways"],"prefix":"10.3390","volume":"28","author":[{"given":"Hong-Yao","family":"Hu","sequence":"first","affiliation":[{"name":"Jilin Medical Products Administration, Changchun 130000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ze-Zhao","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-Ya","family":"Jiang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian-Hua","family":"Duan","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Feng","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin-Guo","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1017\/S2040174409990146","article-title":"Early development and osteoporosis and bone health","volume":"1","author":"Dennison","year":"2010","journal-title":"J. Dev. Orig. Health Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1016\/S2213-8587(21)00119-4","article-title":"Nutritional intake and bone health","volume":"9","author":"Rizzoli","year":"2021","journal-title":"Lancet Diabetes Endocrinol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2243","DOI":"10.1007\/s00198-015-3154-6","article-title":"Burden of high fracture probability worldwide: Secular increases 2010\u20132040","volume":"26","author":"McCloskey","year":"2015","journal-title":"Osteoporos. Int. J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1359\/jbmr.061113","article-title":"Incidence and economic burden of osteoporosis-related fractures in the United States, 2005\u20132025","volume":"22","author":"Burge","year":"2007","journal-title":"J. Bone Miner. Res. Off. J. Am. Soc. Bone Miner. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/S0378-8741(01)00302-6","article-title":"Ameliorative effects of Anoectochilus formosanus extract on osteopenia in ovariectomized rats","volume":"77","author":"Shih","year":"2001","journal-title":"J. Ethnopharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.jep.2005.12.013","article-title":"Effect of ethanol extract of Lepidium meyenii Walp. on osteoporosis in ovariectomized rat","volume":"105","author":"Zhang","year":"2006","journal-title":"J. Ethnopharmacol."},{"key":"ref_7","first-page":"5497046","article-title":"Proteostasis Failure in Neurodegenerative Diseases: Focus on Oxidative Stress","volume":"2020","author":"Tramutola","year":"2020","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"120595","DOI":"10.1016\/j.lfs.2022.120595","article-title":"Mechanisms of autophagy and mitophagy in skeletal development, diseases and therapeutics","volume":"301","author":"Behera","year":"2022","journal-title":"Life Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1038\/s41574-020-0341-0","article-title":"Pathogenesis of glucocorticoid-induced osteoporosis and options for treatment","volume":"16","author":"Chotiyarnwong","year":"2020","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1007\/s11745-014-3950-3","article-title":"High dietary cholesterol masks type 2 diabetes-induced osteopenia and changes in bone microstructure in rats","volume":"49","author":"Lapmanee","year":"2014","journal-title":"Lipids"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Choi, S.S., Lee, S.H., and Lee, K.A. (2022). A Comparative Study of Hesperetin, Hesperidin and Hesperidin Glucoside: Antioxidant, Anti-Inflammatory, and Antibacterial Activities In Vitro. Antioxidants, 11.","DOI":"10.3390\/antiox11081618"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Stanisic, D., Liu, L.H.B., Dos Santos, R.V., Costa, A.F., Dur\u00e1n, N., and Tasic, L. (2020). New Sustainable Process for Hesperidin Isolation and Anti-Ageing Effects of Hesperidin Nanocrystals. Molecules, 25.","DOI":"10.3390\/molecules25194534"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1080\/10408398.2014.906382","article-title":"Health-promoting effects of the citrus flavanone hesperidin","volume":"57","author":"Li","year":"2017","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.lfs.2015.07.014","article-title":"Protective effects of flavonoids against microbes and toxins: The cases of hesperidin and hesperetin","volume":"137","author":"Iranshahi","year":"2015","journal-title":"Life Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s11914-014-0212-5","article-title":"The effects of flavonoids on bone","volume":"12","author":"Welch","year":"2014","journal-title":"Curr. Osteoporos. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1080\/21551197.2012.698220","article-title":"Flavonoid intake and bone health","volume":"31","author":"Weaver","year":"2012","journal-title":"J. Nutr. Gerontol. Geriatr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.ijbiomac.2017.09.014","article-title":"Effects of flavonoids incorporated biological macromolecules based scaffolds in bone tissue engineering","volume":"110","author":"Sanjay","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1892","DOI":"10.1093\/jn\/133.6.1892","article-title":"Hesperidin, a citrus flavonoid, inhibits bone loss and decreases serum and hepatic lipids in ovariectomized mice","volume":"133","author":"Chiba","year":"2003","journal-title":"J. Nutr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1152\/japplphysiol.00441.2007","article-title":"Hesperidin inhibits ovariectomized-induced osteopenia and shows differential effects on bone mass and strength in young and adult intact rats","volume":"104","author":"Horcajada","year":"2008","journal-title":"J. Appl. Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1002\/JPER.16-0708","article-title":"Ameliorative effect of hesperidin on ligation-induced periodontitis in rats","volume":"90","author":"Kuo","year":"2019","journal-title":"J. Periodontol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Miguez, P.A., Tuin, S.A., Robinson, A.G., Belcher, J., Jongwattanapisan, P., Perley, K., de Paiva Gon\u00e7alves, V., Hanifi, A., Pleshko, N., and Barton, E.R. (2021). Hesperidin Promotes Osteogenesis and Modulates Collagen Matrix Organization and Mineralization In Vitro and In Vivo. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22063223"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.2174\/1381612821666150105143358","article-title":"Impact of early life stress on the pathogenesis of mental disorders: Relation to brain oxidative stress","volume":"21","author":"Schiavone","year":"2015","journal-title":"Curr. Pharm. Des."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"8609213","DOI":"10.1155\/2020\/8609213","article-title":"Molecular Mechanisms Linking Oxidative Stress and Diabetes Mellitus","volume":"2020","author":"Yaribeygi","year":"2020","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Cervantes Gracia, K., Llanas-Cornejo, D., and Husi, H. (2017). CVD and Oxidative Stress. J. Clin. Med., 6.","DOI":"10.3390\/jcm6020022"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.jbspin.2012.03.007","article-title":"Reactive oxygen species and exercise on bone metabolism: Friend or enemy?","volume":"79","author":"Filaire","year":"2012","journal-title":"Jt. Bone Spine"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.2106\/JBJS.20.00989","article-title":"Oxidative Stress and Osteoporosis","volume":"103","author":"Kimball","year":"2021","journal-title":"J. Bone Jt. Surg. Am. Vol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5655","DOI":"10.1038\/s41467-022-33008-2","article-title":"Deletion of skeletal muscle Akt1\/2 causes osteosarcopenia and reduces lifespan in mice","volume":"13","author":"Sasako","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.freeradbiomed.2022.12.102","article-title":"17\u03b2-estradiol plays the anti-osteoporosis role via a novel ESR1-Keap1-Nrf2 axis-mediated stress response activation and Tmem119 upregulation","volume":"195","author":"Yang","year":"2023","journal-title":"Free Radic. Biol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Pelusi, L., Mandatori, D., Di Pietrantonio, N., Del Pizzo, F., Di Tomo, P., Di Pietro, N., Buda, R., Genovese, S., Epifano, F., and Pandolfi, A. (2022). Estrogen Receptor 1 (ESR1) and the Wnt\/\u03b2-Catenin Pathway Mediate the Effect of the Coumarin Derivative Umbelliferon on Bone Mineralization. Nutrients, 14.","DOI":"10.3390\/nu14153209"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2025","DOI":"10.1016\/j.biomaterials.2011.11.061","article-title":"The balance of osteogenic and adipogenic differentiation in human mesenchymal stem cells by matrices that mimic stepwise tissue development","volume":"33","author":"Hoshiba","year":"2012","journal-title":"Biomaterials"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1038\/s41401-021-00756-8","article-title":"The m(6)A demethylase FTO promotes the osteogenesis of mesenchymal stem cells by downregulating PPARG","volume":"43","author":"Chen","year":"2022","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Liu, T., Jiang, L., Xiang, Z., Li, J., Zhang, Y., Xiang, T., Wang, W., Li, X., Jia, Y., and Huang, X. (2022). Tereticornate A suppresses RANKL-induced osteoclastogenesis via the downregulation of c-Src and TRAF6 and the inhibition of RANK signaling pathways. Biomed. Pharmacother., 151.","DOI":"10.1016\/j.biopha.2022.113140"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1038\/s41574-022-00649-8","article-title":"Skeletal disorders associated with the growth hormone-insulin-like growth factor 1 axis","volume":"18","author":"Mazziotti","year":"2022","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1172\/JCI72606","article-title":"Vitamin B12-dependent taurine synthesis regulates growth and bone mass","volume":"124","author":"Mottram","year":"2014","journal-title":"J. Clin. Investig."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.tem.2012.03.008","article-title":"Estrogen and the skeleton","volume":"23","author":"Khosla","year":"2012","journal-title":"Trends Endocrinol. Metab. TEM"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2242","DOI":"10.1016\/j.bbrc.2017.12.079","article-title":"Dehydrodiconiferyl alcohol promotes BMP-2-induced osteoblastogenesis through its agonistic effects on estrogen receptor","volume":"495","author":"Lee","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1038\/jcbfm.2012.2","article-title":"Pro-angiogenic effects of resveratrol in brain endothelial cells: Nitric oxide-mediated regulation of vascular endothelial growth factor and metalloproteinases","volume":"32","author":"Pagnussat","year":"2012","journal-title":"J. Cereb. Blood Flow Metab. Off. J. Int. Soc. Cereb. Blood Flow Metab."},{"key":"ref_38","first-page":"11438","article-title":"Relationship between osteogenesis and angiogenesis in ovariectomized osteoporotic rats after exercise training","volume":"10","author":"Li","year":"2017","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.abb.2014.07.006","article-title":"Role of angiogenesis in bone repair","volume":"561","author":"Saran","year":"2014","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1016\/j.ijbiomac.2019.02.022","article-title":"Protective effects of Dipsacus asper polysaccharide on osteoporosis in vivo by regulating RANKL\/RANK\/OPG\/VEGF and PI3K\/Akt\/eNOS pathway","volume":"129","author":"Sun","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_41","first-page":"5985","article-title":"Phosphatidylinositol-3-OH Kinase (PI3K)\/AKT2, activated in breast cancer, regulates and is induced by estrogen receptor alpha (ERalpha) via interaction between ERalpha and PI3K","volume":"61","author":"Sun","year":"2001","journal-title":"Cancer Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4503","DOI":"10.1210\/endo.141.12.7836","article-title":"A role for Akt in mediating the estrogenic functions of epidermal growth factor and insulin-like growth factor I","volume":"141","author":"Martin","year":"2000","journal-title":"Endocrinology"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/19\/6987\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:03:22Z","timestamp":1760130202000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/28\/19\/6987"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,9]]},"references-count":42,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["molecules28196987"],"URL":"https:\/\/doi.org\/10.3390\/molecules28196987","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,9]]}}}