{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:46Z","timestamp":1787028586389,"version":"build-2736575974"},"reference-count":49,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T00:00:00Z","timestamp":1784937600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006705","name":"National Taipei University of Technology","doi-asserted-by":"publisher","award":["NTUT-MMH-No.115-03"],"award-info":[{"award-number":["NTUT-MMH-No.115-03"]}],"id":[{"id":"10.13039\/501100006705","id-type":"DOI","asserted-by":"publisher"}]},{"award":["NTUT-MMH-No.115-03"],"award-info":[{"award-number":["NTUT-MMH-No.115-03"]}],"id":[{"id":"https:\/\/ror.org\/00cn92c09","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>This study aimed to optimize the extraction and purification of TSG-enriched Polygonum multiflorum extract and evaluate its therapeutic potential against androgenetic alopecia. P. multiflorum Thunb. has long been used in traditional Chinese medicine to promote hair growth and preserve hair pigmentation. Microwave-assisted extraction (MAE) was optimized to obtain a 2,3,5,4\u2032-tetrahydroxystilbene-2-O-\u03b2-D-glucoside (TSG)-enriched P. multiflorum extract (PME). The crude extract was further purified by medium-pressure liquid chromatography and semipreparative high-performance liquid chromatography. Phytochemical profiling by HPLC demonstrated a 3.75-fold increase in TSG content, from 22,770.0 \u00b1 815 ppm to 85,387.5 \u00b1 192 ppm, and the identity of the enriched TSG was confirmed by LC\u2013MS and 1H NMR spectroscopy. PME exhibited potent dose-dependent inhibition of 5\u03b1-reductase activity, reaching 92.7% inhibition at 1 \u03bcg\/mL, comparable to that of finasteride. In a testosterone (TES)-induced androgenetic alopecia (AGA) mouse model, PME markedly accelerated hair regrowth and increased both the number and size of hair follicles. Mechanistically, PME suppressed the conversion of TES to dihydrotestosterone (DHT) through 5\u03b1-reductase inhibition and promoted GSK3\u03b2 inactivation, \u03b2-catenin stabilization, and nuclear translocation, indicating activation of the Wnt\/\u03b2-catenin signaling pathway. Consequently, the expression of hair growth-related proteins, including Ki67 (1.98-fold), epidermal growth factor (EGF, 1.54-fold), insulin-like growth factor-1 (IGF-1, 1.21-fold), and vascular endothelial growth factor (VEGF, 1.34-fold), was significantly upregulated. These findings demonstrate, for the first time, that TSG-enriched PME obtained through optimized MAE and chromatographic purification promotes hair regeneration through a dual mechanism involving 5\u03b1-reductase inhibition and Wnt\/\u03b2-catenin activation, highlighting its potential as a natural therapeutic candidate for androgenetic alopecia.<\/jats:p>","DOI":"10.3390\/ijms27156648","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T09:47:41Z","timestamp":1785145661000},"page":"6648","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Optimization and Characterization of TSG-Enriched Polygonum multiflorum Extract and Its Dual Mechanism Against Androgenetic Alopecia via 5\u03b1-Reductase Inhibition and Wnt\/\u03b2-Catenin Activation"],"prefix":"10.3390","volume":"27","author":[{"given":"Te-Yang","family":"Huang","sequence":"first","affiliation":[{"name":"Department of Orthopedic Surgery, Mackay Memorial Hospital, Taipei 104217, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min-Chieh","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei 106344, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7106-6320","authenticated-orcid":false,"given":"Wen-Ta","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei 106344, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"311","DOI":"10.5021\/ad.2012.24.3.311","article-title":"Quality of life assessment in male patients with androgenetic alopecia: Result of a prospective, multicenter study","volume":"24","author":"Han","year":"2012","journal-title":"Ann. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1111\/ics.12657","article-title":"Characteristics of healthy and androgenetic alopecia scalp microbiome: Effect of Lindera strychnifolia roots extract as a natural solution for its modulation","volume":"42","author":"Filaire","year":"2020","journal-title":"Int. J. Cosmet. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.bodyim.2011.06.006","article-title":"Young men\u2019s coping with androgenetic alopecia: Acceptance counts when hair gets thinner","volume":"8","author":"Kranz","year":"2011","journal-title":"Body Image"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Chen, X., Liu, B., Li, Y., Han, L., Tang, X., Deng, W., Lai, W., and Wan, M. (2019). Dihydrotestosterone regulates hair growth through the Wnt\/\u03b2-Catenin Pathway in C57BL\/6 mice and in vitro organ culture. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.01528"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1161\/ATVBAHA.117.309219","article-title":"Testosterone prevents cutaneous ischemia and necrosis in males through complementary estrogenic and androgenic actions","volume":"37","author":"Chenu","year":"2017","journal-title":"Arter. Thromb. Vasc. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/S0090-4295(03)00045-1","article-title":"The role of dihydrotestosterone in benign prostatic hyperplasia","volume":"61","author":"Culley","year":"2003","journal-title":"Urology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0303-7207(02)00372-6","article-title":"Androgens and alopecia","volume":"198","author":"Keith","year":"2002","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1016\/S0531-5565(02)00093-1","article-title":"Molecular mechanisms of androgenetic alopecia","volume":"37","author":"Ralph","year":"2002","journal-title":"Exp. Gerontol."},{"key":"ref_9","first-page":"49","article-title":"Treatment options for androgenetic alopecia","volume":"33","author":"Melissa","year":"2008","journal-title":"US Pharm."},{"key":"ref_10","first-page":"22","article-title":"Topical minoxidil-loaded nanotechnology strategies for alopecia","volume":"2020","author":"Santos","year":"2020","journal-title":"Cosmetic"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e138","DOI":"10.1002\/efd2.138","article-title":"Natural bioactive compounds as potential sources for alleviation and treatment of androgenetic alopecia: A research advance","volume":"5","author":"Fang","year":"2024","journal-title":"eFood"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Elnady, R.E., Abdon, M.S., Shaheen, H.R., Eladawy, R.M., Azar, Y.O., and Al Raish, S.M. (2025). The future of alopecia treatment: Plant extracts, nanocarriers, and 3D bioprinting in focus. Pharmaceutics, 17.","DOI":"10.3390\/pharmaceutics17050584"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Al Raish, S.M., Alsheriafi, H.N., Alkuwaiti, A.A., Safi, S.K., and Safi, A.S. (2026). Insights into knowledge, attitudes, and practices of medicinal plant use in the United Arab Emirates: A cross-sectional study. Front. Nutr., 13.","DOI":"10.3389\/fnut.2026.1755440"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"134","DOI":"10.4103\/wjtcm.wjtcm_78_24","article-title":"Investigation on the therapeutic effect of Polygonum multiflorum Thunb. in chronic stress-induced hair loss in mice coupled with metabolomics and proteomics","volume":"11","author":"Zhao","year":"2025","journal-title":"World J. Tradit. Chin. Med."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Almasri, R.S., Bedir, A.S., and Al Raish, S.M. (2025). Comprehensive Ethnopharmacological Analysis of Medicinal Plants in the UAE: Lawsonia inermis, Nigella sativa, Ziziphus spina-christi, Allium cepa, Allium sativum, Cymbopogon schoenanthus, Matricaria aurea, Phoenix dactylifera, Portulaca oleracea, Reichardia tingitana, Salvadora persica, Solanum lycopersicum, Trigonella foenum-graecum, Withania somnifera, and Ziziphus lotus. Nutrients, 17.","DOI":"10.3390\/nu17030411"},{"key":"ref_16","first-page":"331","article-title":"Poligoni Multiflori Radix enhances osteoblast formation and reduces osteoclast differentiation","volume":"42","author":"Han","year":"2018","journal-title":"Int. J. Mol. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.biopha.2018.01.033","article-title":"Acute toxicity screening of different extractions, components and constituents of Polygonum multiflorum Thunb. on zebrafish (Danio rerio) embryos in vivo","volume":"99","author":"Yang","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.sajb.2018.08.016","article-title":"Maximizing seedling and root tuber production in Polygonum multiflorum for use in ethnomedicine","volume":"119","author":"Kang","year":"2018","journal-title":"S. Afr. J. Bot."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1016\/j.ijbiomac.2018.01.086","article-title":"Preparation and antioxidant activities of important traditional plant polysaccharides","volume":"111","author":"Liu","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.biopha.2012.10.007","article-title":"Proteomic analysis for anti-atherosclerotic effect of tetrahydroxystilbene glucoside in rats","volume":"67","author":"Yao","year":"2013","journal-title":"Biomed. Pharmacother."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"473","DOI":"10.4196\/kjpp.2020.24.6.473","article-title":"2,3,5,4\u2032-Tetrahydroxystilbene-2-O-\u03b2-D-Glucoside modulated human umbilical vein endothelial cells injury under oxidative stress","volume":"24","author":"Guo","year":"2020","journal-title":"Korean J. Physiol. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.ejphar.2007.09.013","article-title":"Protective effects of 2,3,5,4\u2032-tetrahydroxystilbene-2-O-beta-d-glucoside, an active component of Polygonum multiflorum Thunb, on experimental colitis in mice","volume":"578","author":"Wang","year":"2008","journal-title":"Eur. J. Pharmacol."},{"key":"ref_23","first-page":"1380146","article-title":"Tetrahydroxystilbene glucoside effectively prevents apoptosis induced hair loss","volume":"2018","author":"Chen","year":"2018","journal-title":"BioMed Res. Int."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Qian, J., Feng, C., Wu, Z., Yang, Y., Gao, X., Zhu, L., Liu, Y., and Gao, Y. (2024). Phytochemistry, pharmacology, toxicology and detoxification of Polygonum multiflorum Thunb.: A comprehensive review. Front. Pharmacol., 15.","DOI":"10.3389\/fphar.2024.1427019"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.jhip.2025.12.005","article-title":"Research progress on the application of Pleuropterus multiflorus in the treatment of androgenetic alopecia","volume":"6","author":"Han","year":"2025","journal-title":"J. Holist. Integr. Pharm."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.jbiosc.2020.08.007","article-title":"Phosphorylation of levan by microwave-assisted synthesis enhanced anticancerability","volume":"131","author":"Huang","year":"2021","journal-title":"J. Biosci. Bioeng."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1016\/j.seppur.2008.03.025","article-title":"Microwave-assisted extraction of flavonoids from Radix astragali","volume":"62","author":"Xiao","year":"2008","journal-title":"Sep. Purif. Technol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"277","DOI":"10.5124\/jkma.2020.63.5.277","article-title":"Pharmacological therapeutics in androgenetic alopecia","volume":"63","author":"Kim","year":"2020","journal-title":"J. Korean Med. Assoc."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Varghese, F., Bukhari, A.B., Malhotra, R., and De, A. (2014). IHC Profiler: An open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0096801"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"120627","DOI":"10.1016\/j.jep.2025.120627","article-title":"Tetrahydroxy stilbene glucoside promotes hair regeneration by inducing Th22 cell differentiation","volume":"355","author":"Zhang","year":"2026","journal-title":"J. Ethnopharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/j.jep.2011.03.028","article-title":"Topical application of Polygonum multiflorum extract induces hair growth of resting hair follicles through upregulating Shh and \u03b2-catenin expression in C57BL\/6 mice","volume":"135","author":"Park","year":"2011","journal-title":"J. Ethnopharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Shin, J.Y., Choi, Y.H., Kim, J., Park, S.Y., Nam, Y.J., Lee, S.Y., Jeon, J.H., Jin, M.H., and Lee, S. (2020). Polygonum multiflorum extract support hair growth by elongating anagen phase and abrogating the effect of androgen in cultured human dermal papilla cells. BMC Complement. Med. Ther., 20.","DOI":"10.1186\/s12906-020-02940-5"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1002\/(SICI)1097-0045(19980301)34:4<283::AID-PROS6>3.0.CO;2-I","article-title":"Presence of 5\u03b1-Reductase isozymes and aromatase in human prostate cancer cells and in benign prostate hyperplastic tissue","volume":"34","author":"Poletti","year":"1998","journal-title":"Prostate"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.fitote.2016.08.008","article-title":"Mechanism of action of herbs and their active constituents used in hair loss treatment","volume":"114","author":"Herman","year":"2016","journal-title":"Fitoterapia"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Truong, V.L., Bak, M.J., Lee, C., Jun, M., and Jeong, W.S. (2017). Hair regenerative mechanisms of red ginseng oil and its major components in the testosterone-induced delay of anagen entry in C57BL\/6 mice. Molecules, 22.","DOI":"10.3390\/molecules22091505"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Lin, Y., Liu, C., Zhan, X., Wang, B., Li, K., and Li, J. (2020). Jagged1 and epidermal growth factor promoted androgen- suppressed mouse hair growth in vitro and in vivo. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.01634"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.jgr.2019.05.013","article-title":"Effects of a gintonin-enriched fraction on hair growth: An in vitro and in vivo study","volume":"44","author":"Lee","year":"2020","journal-title":"J. Ginseng Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/s40543-019-0178-0","article-title":"The effect of Lactobacillus plantarum hydrolysates promoting VEGF production on vascular growth and hair growth of C57BL\/6 mice","volume":"10","author":"Woo","year":"2019","journal-title":"J. Anal. Sci. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1798","DOI":"10.1161\/CIRCRESAHA.110.219840","article-title":"An Updated overview on Wnt signaling pathways","volume":"106","author":"Tata","year":"2010","journal-title":"Circ. Res."},{"key":"ref_40","first-page":"1934578X20951433","article-title":"Hair growth is promoted in mouse dorsal skin by a mixture of Platycladus orientalis (L.) Franco Leaf extract and alpha-terpineol by increasing growth factors and wnt3\/\u03b2-Catenin","volume":"15","author":"Ahn","year":"2020","journal-title":"Nat. Prod. Commun."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"354","DOI":"10.4062\/biomolther.2019.206","article-title":"Vanillic acid stimulates anagen signaling via the PI3K\/Akt\/\u03b2-Catenin pathway in dermal papilla cells","volume":"28","author":"Kang","year":"2020","journal-title":"Biomol. Ther."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e43025","DOI":"10.1097\/MD.0000000000043025","article-title":"Exploring the molecular mechanism of Polygonum multiflorum in treating androgenic alopecia based on the methods of bioinformatics and molecular docking","volume":"104","author":"Zhong","year":"2025","journal-title":"Medicine"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhu, C., Li, J., Tang, W., Li, Y., Lin, C., Peng, D., and Yang, C. (2025). 2,3,5,4\u2032-Tetrahydroxystilbene-2-O-\u03b2-D-glucoside (TSG) from Polygonum multiflorum Thunb.: A Systematic Review on Anti-Aging. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26073381"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"517901","DOI":"10.1155\/2015\/517901","article-title":"Hair Growth Promotion Activity and Its Mechanism of Polygonum multiflorum","volume":"2015","author":"Li","year":"2015","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Ho, C.Y., Chen, J.Y.F., Hsu, W.L., Yu, S., Chen, W.C., Chiu, S.H., Yang, H.R., Lin, S.Y., and Wu, C.Y. (2023). Female pattern hair loss: An overview with focus on the genetics. Genes, 14.","DOI":"10.3390\/genes14071326"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"174","DOI":"10.4103\/1027-8117.357357","article-title":"Comorbid laboratory abnormalities in female pattern hair loss patients","volume":"40","author":"Wang","year":"2022","journal-title":"Dermatol. Sin."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Chaiyana, W., Punyoyai, C., Somwongin, S., Leelapornpisid, P., Ingkaninan, K., Waranuch, N., Srivilai, J., Thitipramote, N., Wisuitiprot, W., and Schuster, R. (2017). Inhibition of 5\u03b1-reductase, IL-6 secretion, and oxidation process of Equisetum debile Roxb. ex. vaucher extract as functional food and nutraceuticals ingredients. Nutrients, 9.","DOI":"10.3390\/nu9101105"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1248\/bpb.b18-00283","article-title":"Screening of Euphorbiaceae plant extracts for anti-5\u03b1-reductase","volume":"41","author":"Kamei","year":"2018","journal-title":"Biol. Pharm. Bull."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1002\/jvc2.86","article-title":"Mapping the hair density, thickness, and volume in normal and androgenetic alopecia subjects with the digital microscope","volume":"2","author":"Chang","year":"2023","journal-title":"JEADV Clin. Pract."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6648\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T04:31:21Z","timestamp":1785299481000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6648"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,25]]},"references-count":49,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27156648"],"URL":"https:\/\/doi.org\/10.3390\/ijms27156648","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,25]]}}}