{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:13:01Z","timestamp":1792789981758,"version":"4.5.0"},"reference-count":35,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T00:00:00Z","timestamp":1792022400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T00:00:00Z","timestamp":1792022400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003715","name":"Korea Research Institute of Bioscience and Biotechnology","doi-asserted-by":"publisher","award":["KGM1052612"],"award-info":[{"award-number":["KGM1052612"]}],"id":[{"id":"10.13039\/501100003715","id-type":"DOI","asserted-by":"publisher"}]},{"award":["KGM1052612"],"award-info":[{"award-number":["KGM1052612"]}],"id":[{"id":"https:\/\/ror.org\/03ep23f07","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Ministry of Science and ICT","award":["2710085200"],"award-info":[{"award-number":["2710085200"]}],"id":[{"id":"https:\/\/ror.org\/01wpjm123","id-type":"ROR","asserted-by":"crossref"}]},{"award":["2710085200"],"award-info":[{"award-number":["2710085200"]}],"id":[{"id":"https:\/\/ror.org\/01wpjm123","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background: Phytoestrogens are promising alternatives to menopausal hormone therapy, but their clinical application is limited by poor solubility and bioavailability. Silybum marianum (milk thistle) contains flavonolignans with reported estrogenic potential. However, its extract shows only modest estrogenic activity, and approaches to enhance its efficacy by improving solubility remain insufficiently explored. Methods: A water-soluble Silybum marianum complex (WS) was prepared using microwave-assisted processing with steviol glycoside (SG), and its phytochemical integrity was assessed by UPLC-QTOF-MS. Estrogenic activity was evaluated using cell-free ER\u03b1\/ER\u03b2 competitive binding assays and, in MCF-7 cells, by assessing proliferation and ER\u03b1\/ERK\/AKT phosphorylation. In vivo efficacy was evaluated in a VCD-induced menopause model in female C57BL\/6 mice by measuring plasma estradiol levels, uterine weight, and vaginal epithelial thickness. Results: WS retained major flavonolignans and incorporated SG-derived metabolites. WS bound both ER\u03b1 and ER\u03b2 in cell-free assays. In MCF-7 cells, WS induced ER\u03b1, ERK, and AKT phosphorylation and cell proliferation; the proliferative response and ERK\/AKT phosphorylation were abolished by the ER antagonist fulvestrant, and proliferation was comparable to that of genistein and daidzein. Neither Silybum marianum extract (SME) alone nor its simple physical combinations with SG reproduced the activity of WS, indicating that enhanced estrogenic activity was observed following microwave-assisted processing. In vivo, WS dose-dependently increased measured estradiol levels, increased uterine weight, and reversed vaginal atrophy. Conclusions: Microwave-assisted processing with SG significantly enhances the estrogenic activity of Silybum marianum. Our findings suggest that WS may serve as a promising phytoestrogenic candidate for further evaluation in the context of estrogen deficiency.<\/jats:p>","DOI":"10.3390\/nu18203388","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T16:56:13Z","timestamp":1792083373000},"page":"3388","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enhanced Estrogenic Activity in Water-Soluble Silybum marianum Complex via Microwave-Assisted Steviol Glycoside-Based Formulation"],"prefix":"10.3390","volume":"18","author":[{"given":"Ju-Hong","family":"Kim","sequence":"first","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"},{"name":"Laboratory of Veterinary Pathology, College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of 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Hwan","family":"Jeong","sequence":"additional","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Ji-Young","family":"Park","sequence":"additional","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Hyung Jae","family":"Jeong","sequence":"additional","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8748-9731","authenticated-orcid":false,"given":"In-Chul","family":"Lee","sequence":"additional","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0196-4672","authenticated-orcid":false,"given":"Hyung-Jun","family":"Kwon","sequence":"additional","affiliation":[{"name":"Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of 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