{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:54:09Z","timestamp":1785405249869,"version":"3.56.0"},"reference-count":40,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T00:00:00Z","timestamp":1774915200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF-2021R1A2C2013483"],"award-info":[{"award-number":["NRF-2021R1A2C2013483"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"award":["NRF-2021R1A2C2013483"],"award-info":[{"award-number":["NRF-2021R1A2C2013483"]}],"id":[{"id":"https:\/\/ror.org\/013aysd81","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Osteoporosis is a progressive systemic skeletal disease, with male osteoporosis emerging as a critical global concern due to high morbidity and mortality from fractures. This study investigated the anti-osteoporotic potential of CGAC\u2014a herbal mixture of Cervus elaphus Linnaeus, Glycine max (L.) Merr., Angelica gigas Nakai, and Cnidium officinale Makino\u2014and its underlying mechanisms in an orchiectomized (ORX) mouse model. Methods: C57BL\/6J mice underwent ORX for 8 weeks, followed by CGAC administration (250 and 500 mg\/kg) for an additional 8 weeks. Molecular mechanisms were further validated using MG63 osteoblastic and RAW 264.7 osteoclast assays. Results: ORX induced severe osteoporotic phenotypes, including significant reductions in bone mineral density (BMD) and trabecular microarchitecture. Notably, at the time point examined, ORX was associated with a suppressed bone remodeling state, reflected by reductions in both TRAP-positive osteoclasts and ALP-positive osteoblasts, together with lower serum BALP, CTX-1, and Gla\/Glu-OC ratio. Conversely, CGAC normalized this stagnant state and restored physiological remodeling. This was accompanied by reduced marrow fat accumulation through the AMPK signaling axis, which upregulated Runx2 and downregulated PPAR-\u03b3. In vitro results confirmed that CGAC promoted osteoblast differentiation and mineralization while suppressing RANKL-induced osteoclastogenesis. These actions suggest that CGAC may be involved in regulating Wnt\/\u03b2-catenin signaling. Conclusions: Overall, CGAC is a promising therapeutic candidate for male osteoporosis, offering pharmacological benefits particularly relevant to aging populations.<\/jats:p>","DOI":"10.3390\/ph19040555","type":"journal-article","created":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T09:26:12Z","timestamp":1775121972000},"page":"555","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Standard Herbal Formula, CGAC, Attenuates Bone Loss by Normalizing Low-Bone Turnover Stagnation in an Orchiectomy-Induced Mouse Model"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4523-8920","authenticated-orcid":false,"given":"Dong-Cheol","family":"Baek","sequence":"first","affiliation":[{"name":"Institute of Bioscience & Integrative Medicine, Daejeon University, Daejeon 35235, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7086-1596","authenticated-orcid":false,"given":"Min-Young","family":"Chae","sequence":"additional","affiliation":[{"name":"Department of Korean Rehabilitation Medicine, College of Korean Medicine, Daejeon University, Daejeon 35235, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1311-829X","authenticated-orcid":false,"given":"Tae-Wook","family":"Woo","sequence":"additional","affiliation":[{"name":"Institute of Bioscience & Integrative Medicine, Daejeon University, Daejeon 35235, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4876-0167","authenticated-orcid":false,"given":"Chang-Gue","family":"Son","sequence":"additional","affiliation":[{"name":"Institute of Bioscience & Integrative Medicine, Daejeon University, Daejeon 35235, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9468-9694","authenticated-orcid":false,"given":"Eun-Jung","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Korean Rehabilitation Medicine, College of Korean Medicine, Daejeon University, Daejeon 35235, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,31]]},"reference":[{"key":"ref_1","first-page":"1017","article-title":"Osteoporosis: Pathophysiology and therapeutic options","volume":"19","author":"Dovjak","year":"2020","journal-title":"EXCLI J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"101766","DOI":"10.1016\/j.berh.2022.101766","article-title":"Male osteoporosis-what are the causes, diagnostic challenges, and management","volume":"36","author":"Clarke","year":"2022","journal-title":"Best Pract. Res. Clin. Rheumatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1007\/s11657-018-0540-7","article-title":"Prevalence of osteoporosis with the World Health Organization diagnostic criteria in the Eastern Mediterranean Region: A systematic review and meta-analysis","volume":"13","author":"Zamani","year":"2018","journal-title":"Arch. Osteoporos."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Chen, J.-F., Lin, P.-W., Tsai, Y.-R., Yang, Y.-C., and Kang, H.-Y. (2019). Androgens and androgen receptor actions on bone health and disease: From androgen deficiency to androgen therapy. Cells, 8.","DOI":"10.3390\/cells8111318"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1172\/JCI10942","article-title":"Relative contributions of testosterone and estrogen in regulating bone resorption and formation in normal elderly men","volume":"106","author":"Riggs","year":"2000","journal-title":"J. Clin. Investig."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1016\/j.bone.2009.10.039","article-title":"Signaling pathways implicated in androgen regulation of endocortical bone","volume":"46","author":"Wiren","year":"2010","journal-title":"Bone"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1210\/jcem.87.2.8201","article-title":"Age trends in the level of serum testosterone and other hormones in middle-aged men: Longitudinal results from the Massachusetts male aging study","volume":"87","author":"Feldman","year":"2002","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1552","DOI":"10.1359\/jbmr.080518","article-title":"Older men with low serum estradiol and high serum SHBG have an increased risk of fractures","volume":"23","author":"Vandenput","year":"2008","journal-title":"J. Bone Miner. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2049","DOI":"10.1007\/s00198-021-05900-y","article-title":"The clinician\u2019s guide to prevention and treatment of osteoporosis","volume":"33","author":"LeBoff","year":"2022","journal-title":"Osteoporos. Int."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2355","DOI":"10.1007\/s00198-015-3148-4","article-title":"Efficacy of bisphosphonates against osteoporosis in adult men: A meta-analysis of randomized controlled trials","volume":"26","author":"Chen","year":"2015","journal-title":"Osteoporos. Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/1759720X13477714","article-title":"Bisphosphonate drug holiday: Who, when and how long","volume":"5","author":"Diab","year":"2013","journal-title":"Ther. Adv. Musculoskelet. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1093\/jbmr\/zjaf037","article-title":"Denosumab discontinuation in the clinic: Implications of rebound bone turnover and emerging strategies to prevent bone loss and fractures","volume":"40","author":"Kumar","year":"2025","journal-title":"J. Bone Miner. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"104717","DOI":"10.1016\/j.bse.2023.104717","article-title":"Angelica gigas Nakai: An overview on its chemical composition and pharmacological activity","volume":"111","author":"He","year":"2023","journal-title":"Biochem. Syst. Ecol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3309","DOI":"10.1080\/23802359.2019.1673227","article-title":"Characterization of the complete chloroplast genome of black soybeans, Glycine max (L.) Merr. (legume)","volume":"4","author":"Jin","year":"2019","journal-title":"Mitochondrial DNA Part B"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"116052","DOI":"10.1016\/j.jep.2022.116052","article-title":"Pilose antler (Cervus elaphus Linnaeus) polysaccharide and polypeptide extract inhibits bone resorption in high turnover type osteoporosis by stimulating the MAKP and MMP-9 signaling pathways","volume":"304","author":"Liu","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e202301639","DOI":"10.1002\/cbdv.202301639","article-title":"Cnidium officinale Makino: Phytology, Phytochemistry, Toxicology, Pharmacology and Prescriptions (1967\u20132023)","volume":"21","author":"Ma","year":"2024","journal-title":"Chem. Biodivers."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Baek, D.-C., Hwang, S.-J., Lee, J.-S., Wang, J.-H., Son, C.-G., and Lee, E.-J. (2023). A Mixture of Cervus elaphus sibiricus and Glycine max (L.) Merrill Inhibits Ovariectomy-Induced Bone Loss Via Regulation of Osteogenic Molecules in a Mouse Model. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24054876"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1089\/jmf.2011.2047","article-title":"Ultrafine Angelica gigas powder normalizes ovarian hormone levels and has antiosteoporosis properties in ovariectomized rats: Particle size effect","volume":"15","author":"Choi","year":"2012","journal-title":"J. Med. Food"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lee, K.-Y., Kim, J.-H., Kim, E.-Y., Yeom, M., Jung, H.-S., and Sohn, Y. (2019). Water extract of Cnidii Rhizoma suppresses RANKL-induced osteoclastogenesis in RAW 264.7 cell by inhibiting NFATc1\/c-Fos signaling and prevents ovariectomized bone loss in SD-rat. BMC Complement. Altern. Med., 19.","DOI":"10.1186\/s12906-019-2611-8"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e71378","DOI":"10.1002\/hsr2.71378","article-title":"Global Health Effects of Osteopenia and Osteoporosis, 1990\u20132021","volume":"8","author":"Safiri","year":"2025","journal-title":"Health Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1038\/bonekey.2014.109","article-title":"Rodent models of osteoporosis","volume":"3","author":"Sophocleous","year":"2014","journal-title":"BoneKEy Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1055\/s-0034-1367031","article-title":"Testosterone regulates bone response to inflammation","volume":"46","author":"Steffens","year":"2014","journal-title":"Horm. Metab. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1589","DOI":"10.1016\/j.cell.2025.01.006","article-title":"Identification, structure, and agonist design of an androgen membrane receptor","volume":"188","author":"Yang","year":"2025","journal-title":"Cell"},{"key":"ref_24","first-page":"35","article-title":"Trabecular bone microarchitecture and male osteoporosis","volume":"83","author":"Legrand","year":"1999","journal-title":"Morphol. Bull. L\u2019association Anat."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1152\/physrev.00033.2015","article-title":"Estrogens and androgens in skeletal physiology and pathophysiology","volume":"97","author":"Almeida","year":"2016","journal-title":"Physiol. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Schwartz, A.V. (2015). Marrow fat and bone: Review of clinical findings. Front. Endocrinol., 6.","DOI":"10.3389\/fendo.2015.00040"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1530\/JOE-11-0306","article-title":"AMP-activated protein kinase pathway and bone metabolism","volume":"212","author":"Jeyabalan","year":"2011","journal-title":"J. Endocrinol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.freeradbiomed.2023.05.033","article-title":"AMPK negatively regulates RANKL-induced osteoclast differentiation by controlling oxidative stress","volume":"205","author":"Tanaka","year":"2023","journal-title":"Free Radic. Biol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"105379","DOI":"10.1016\/j.jbc.2023.105379","article-title":"AMPK\u03b11 negatively regulates osteoclastogenesis and mitigates pathological bone loss","volume":"299","author":"Ribeiro","year":"2023","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1210\/jcem-69-4-776","article-title":"Increases in bone density during treatment of men with idiopathic hypogonadotropic hypogonadism","volume":"69","author":"Finkelstein","year":"1989","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Gao, J., Wang, Z., Gao, P., Fan, Q., Zhang, T., Cui, L., Shi, L., Liu, Z., Yang, Z., and He, L. (2022). Daphnetin alleviates senile and disuse osteoporosis by distinct modulations of bone formation and resorption. Antioxidants, 11.","DOI":"10.3390\/antiox11122365"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1038\/s41421-024-00689-6","article-title":"Cell signaling and transcriptional regulation of osteoblast lineage commitment, differentiation, bone formation, and homeostasis","volume":"10","author":"Zhu","year":"2024","journal-title":"Cell Discov."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/j.maturitas.2014.04.013","article-title":"Wnt signaling and osteoporosis","volume":"78","author":"Manolagas","year":"2014","journal-title":"Maturitas"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3071","DOI":"10.1128\/MCB.01428-09","article-title":"Osteocyte Wnt\/\u03b2-catenin signaling is required for normal bone homeostasis","volume":"30","author":"Kramer","year":"2010","journal-title":"Mol. Cell. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Vasiliadis, E.S., Evangelopoulos, D.-S., Kaspiris, A., Benetos, I.S., Vlachos, C., and Pneumaticos, S.G. (2022). The role of sclerostin in bone diseases. J. Clin. Med., 11.","DOI":"10.3390\/jcm11030806"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"18667","DOI":"10.1002\/jcb.29153","article-title":"Ligustilide suppresses RANKL-induced osteoclastogenesis and bone resorption via inhibition of RANK expression","volume":"120","author":"Wang","year":"2019","journal-title":"J. Cell. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1186\/s13018-024-05371-x","article-title":"Tetramethylpyrazine promotes osteo-angiogenesis during bone fracture repair","volume":"20","author":"Ai","year":"2025","journal-title":"J. Orthop. Surg. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1186\/s13020-022-00653-0","article-title":"Isoflavones daidzin and daidzein inhibit lipopolysaccharide-induced inflammation in RAW264. 7 macrophages","volume":"17","author":"Tan","year":"2022","journal-title":"Chin. Med."},{"key":"ref_39","first-page":"e59270","article-title":"Murine orchiectomy and ovariectomy to reduce sex hormone production","volume":"147","author":"Rowe","year":"2023","journal-title":"J. Vis. Exp. JoVE"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1186\/scrt328","article-title":"Effects of dexamethasone, ascorbic acid and \u03b2-glycerophosphate on the osteogenic differentiation of stem cells in vitro","volume":"4","author":"Langenbach","year":"2013","journal-title":"Stem Cell Res. Ther."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/555\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T09:34:51Z","timestamp":1775122491000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/555"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,31]]},"references-count":40,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ph19040555"],"URL":"https:\/\/doi.org\/10.3390\/ph19040555","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,31]]}}}