{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:12:28Z","timestamp":1785420748991,"version":"3.56.0"},"reference-count":31,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2020YFA0803801"],"award-info":[{"award-number":["2020YFA0803801"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFC3502002"],"award-info":[{"award-number":["2022YFC3502002"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U23A20500"],"award-info":[{"award-number":["U23A20500"]}],"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":["81873568"],"award-info":[{"award-number":["81873568"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["U23A20500"],"award-info":[{"award-number":["U23A20500"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["81873568"],"award-info":[{"award-number":["81873568"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background: Postmenopausal osteoporosis is a common metabolic bone disorder characterized by decreased bone mass and microstructural deterioration, often accompanied by increased bone marrow adiposity and systemic fat accumulation. Kun-Ling Wan Formula (KLW) is a compound Chinese medicine clinically used for gynecological disorders, though its effects on postmenopausal osteoporosis and associated fat accumulation remain unclear. Distinct from previous herbal formulation studies that primarily focused on bone outcomes, our study uniquely integrates bone protection, marrow adiposity reduction, systemic metabolic improvement, and multi-omics mechanistic dissection in a high-fat diet-fed ovariectomized mouse model. Methods: KLW chemical composition was analyzed by UPLC-Q-TOF\/MS. Ovariectomized (OVX) C57BL\/6J mice fed high-fat or normal diet were treated with KLW at clinically equivalent or double doses, with estrogen and active compounds as controls. Bone microstructure was assessed by micro-CT, bone marrow fat by MRI-PDFF, and metabolism by OGTT, ITT, and metabolic cages. Network pharmacology, proteomics, molecular docking, and dynamics simulations identified core targets. C3H10T1\/2 cells were used to assess osteogenic\/adipogenic differentiation and mTOR pathway activation. Results: Twelve compounds were identified in KLW. In OVX mice, KLW significantly improved bone mineral density and trabecular microstructure, reduced adiposity and bone marrow fat, and enhanced glucose tolerance and insulin sensitivity. In vitro, KLW promoted osteogenesis and suppressed adipogenesis in C3H10T1\/2 cells. Integrative analyses identified mTOR as a central target, with chrysophanol, pyrogallol, and apigenin showing high-affinity binding. KLW inhibited mTOR\/S6K phosphorylation during differentiation, an effect reversible by leucine. Conclusions: KLW ameliorates osteoporosis and reduces fat accumulation in OVX mice by shifting mesenchymal stem cell differentiation toward osteogenesis via mTOR pathway modulation.<\/jats:p>","DOI":"10.3390\/ph19050719","type":"journal-article","created":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T10:15:27Z","timestamp":1777630527000},"page":"719","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Kun-Ling Wan Formula Ameliorates Postmenopausal Osteoporosis and Adipose Accumulation by Suppressing mTOR Signaling in Mesenchymal Stem Cells"],"prefix":"10.3390","volume":"19","author":[{"given":"Xiaoqing","family":"Lu","sequence":"first","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He","family":"Lan","sequence":"additional","affiliation":[{"name":"Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing 100191, China"},{"name":"Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing 100053, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaqi","family":"Fan","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Yang","sequence":"additional","affiliation":[{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qianzan","family":"Liao","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0801-4574","authenticated-orcid":false,"given":"Yaoxuan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingxin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dexin","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunshui","family":"Pan","sequence":"additional","affiliation":[{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quan","family":"Li","sequence":"additional","affiliation":[{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Sun","sequence":"additional","affiliation":[{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinmei","family":"Huo","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Yuwen","sequence":"additional","affiliation":[{"name":"National Institute on Drug Dependence and Beijing Key Laboratory of Drug Dependence Research, Peking University, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing-Yan","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4019-3391","authenticated-orcid":false,"given":"Yin","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Haidian District, Beijing 100191, China"},{"name":"Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"a031211","DOI":"10.1101\/cshperspect.a031211","article-title":"Regulation of Bone Metabolism by Sex Steroids","volume":"8","author":"Khosla","year":"2018","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1080\/13697137.2021.1951206","article-title":"Osteoporosis and fractures in women: The burden of disease","volume":"25","author":"Lorentzon","year":"2022","journal-title":"Climacteric"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1929","DOI":"10.1007\/s00198-015-3093-2","article-title":"Projection of osteoporosis-related fractures and costs in China: 2010\u20132050","volume":"26","author":"Si","year":"2015","journal-title":"Osteoporos. Int."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.cytogfr.2020.02.003","article-title":"The relationship between bone marrow adipose tissue and bone metabolism in postmenopausal osteoporosis","volume":"52","author":"Li","year":"2020","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1177\/08971900231167925","article-title":"The Art of Hormone Replacement Therapy (HRT) in Menopause Management","volume":"37","author":"Cameron","year":"2024","journal-title":"J. Pharm. Pract."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"101577","DOI":"10.1016\/j.beem.2021.101577","article-title":"Menopausal hormone therapy and breast cancer risk","volume":"35","author":"Rozenberg","year":"2021","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_7","unstructured":"Valdes, A., Patel, P., and Bajaj, T. (2025). Estrogen Therapy. StatPearls, StatPearls Publishing."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1080\/13697137.2025.2503874","article-title":"Safety of menopause hormone therapy in postmenopausal women at higher risk of venous thromboembolism: A systematic review","volume":"28","author":"Hicks","year":"2025","journal-title":"Climacteric"},{"key":"ref_9","first-page":"67","article-title":"Alpha-lipoic acid in pharmaceutical development: A comprehensive review of its therapeutic potential and molecular mechanisms","volume":"23","author":"Koyuncu","year":"2025","journal-title":"Prospect. Pharm. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Tong, Y., Tu, Y., Wang, J., Liu, X., Su, Q., Wang, Y., and Wang, W. (2025). Mechanisms and therapeutic strategies linking mesenchymal stem cells senescence to osteoporosis. Front. Endocrinol., 16.","DOI":"10.3389\/fendo.2025.1625806"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Chen, R., Yang, C., Yang, F., Yang, A., Xiao, H., Peng, B., Chen, C., Geng, B., and Xia, Y. (2024). Targeting the mTOR-Autophagy Axis: Unveiling Therapeutic Potentials in Osteoporosis. Biomolecules, 14.","DOI":"10.3390\/biom14111452"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Xu, Y., Pan, C.S., Li, Q., Zhang, H.L., Yan, L., Anwaier, G., Wang, X.Y., Yan, L.L., Fan, J.Y., and Li, D. (2021). The Ameliorating Effects of Bushen Huatan Granules and Kunling Wan on Polycystic Ovary Syndrome Induced by Dehydroepiandrosterone in Rats. Front. Physiol., 12.","DOI":"10.3389\/fphys.2021.525145"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Ren, Y., Song, X., Tan, L., Guo, C., Wang, M., Liu, H., Cao, Z., Li, Y., and Peng, C. (2020). A Review of the Pharmacological Properties of Psoralen. Front. Pharmacol., 11.","DOI":"10.3389\/fphar.2020.571535"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110166","DOI":"10.1016\/j.plaphy.2025.110166","article-title":"Identification and catalytic mechanisms of key enzymes in the biosynthesis of asperosaponin VI from Dipsacusasperoides","volume":"227","author":"Yang","year":"2025","journal-title":"Plant Physiol. Biochem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"116426","DOI":"10.1016\/j.jep.2023.116426","article-title":"Psoralen prevents the inactivation of estradiol and treats osteoporosis via covalently targeting HSD17B2","volume":"311","author":"Lu","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref_16","first-page":"1156","article-title":"The osteoprotective effect of psoralen in ovariectomy-induced osteoporotic rats via stimulating the osteoblastic differentiation from bone mesenchymal stem cells","volume":"19","author":"Yang","year":"2012","journal-title":"Menopause"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"368","DOI":"10.3892\/etm.2021.9799","article-title":"Psoralen, a natural phytoestrogen, improves diaphyseal fracture healing in ovariectomized mice: A preliminary study","volume":"21","author":"Huang","year":"2021","journal-title":"Exp. Ther. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"940","DOI":"10.3892\/etm.2021.10372","article-title":"Psoralen accelerates osteogenic differentiation of human bone marrow mesenchymal stem cells by activating the TGF-\u03b2\/Smad3 pathway","volume":"22","author":"Huang","year":"2021","journal-title":"Exp. Ther. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.jare.2024.11.036","article-title":"Asperosaponin VI inhibition of DNMT alleviates GPX4 suppression-mediated osteoblast ferroptosis and diabetic osteoporosis","volume":"75","author":"Wei","year":"2024","journal-title":"J. Adv. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1007\/s00223-022-01057-w","article-title":"Asperosaponin VI Protects Against Bone Loss Due to Hindlimb Unloading in Skeletally Growing Mice Through Regulating Microbial Dysbiosis Altering the 5-HT Pathway","volume":"112","author":"Niu","year":"2023","journal-title":"Calcif. Tissue Int."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1038\/s41580-019-0199-y","article-title":"mTOR at the nexus of nutrition, growth, ageing and disease","volume":"21","author":"Liu","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41413-017-0004-5","article-title":"mTOR signaling in skeletal development and disease","volume":"6","author":"Chen","year":"2018","journal-title":"Bone Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.cmet.2017.07.001","article-title":"The Dawn of the Age of Amino Acid Sensors for the mTORC1 Pathway","volume":"26","author":"Wolfson","year":"2017","journal-title":"Cell Metab."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"104261","DOI":"10.1016\/j.fbio.2024.104261","article-title":"The relationships between high-fat diet and metabolic syndrome: Potential mechanisms","volume":"59","author":"Tang","year":"2024","journal-title":"Food Biosci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e230116","DOI":"10.1530\/JOE-23-0116","article-title":"Postmenopausal osteoporosis coexisting with sarcopenia: The role and mechanisms of estrogen","volume":"259","author":"Lu","year":"2023","journal-title":"J. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1093\/abbs\/gmr041","article-title":"mTOR and the differentiation of mesenchymal stem cells","volume":"43","author":"Xiang","year":"2011","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1002\/ddr.21461","article-title":"Dose translation between laboratory animals and human in preclinical and clinical phases of drug development","volume":"79","author":"Nair","year":"2018","journal-title":"Drug Dev. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1186\/s12967-023-04233-0","article-title":"Uncovering the mechanism of resveratrol in the treatment of diabetic kidney disease based on network pharmacology, molecular docking, and experimental validation","volume":"21","author":"Chen","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"D605","DOI":"10.1093\/nar\/gkaa1074","article-title":"The STRING database in 2021: Customizable protein-protein networks, and functional characterization of user-uploaded gene\/measurement sets","volume":"49","author":"Szklarczyk","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.biosystems.2014.11.005","article-title":"CytoNCA: A cytoscape plugin for centrality analysis and evaluation of protein interaction networks","volume":"127","author":"Tang","year":"2015","journal-title":"Biosystems"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"R60","DOI":"10.1186\/gb-2003-4-9-r60","article-title":"DAVID: Database for Annotation, Visualization, and Integrated Discovery","volume":"4","author":"Dennis","year":"2003","journal-title":"Genome Biol."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/5\/719\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T04:12:42Z","timestamp":1778731962000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/5\/719"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,30]]},"references-count":31,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["ph19050719"],"URL":"https:\/\/doi.org\/10.3390\/ph19050719","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,30]]}}}