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SMAD2\/3 signal transduction is known to play a crucial role in osteogenic differentiation through transforming growth factor-beta (TGF-\u03b2). By screening a library of small-molecule compounds, the current study identifies higenamine (HG) as an active osteogenic agent that could be a therapeutic candidate for osteoporosis. In vitro data demonstrated that HG effectively induced expressions of osteogenic markers in mouse bone marrow stromal cell (BMSCs) and preosteoblastic cell cultures. Further, HG treatment resulted in enhanced bone formation and prevented accelerated bone loss on two animal models that mimic spontaneous senile osteoporosis and postmenopausal osteoporosis. IQ motif-containing GTPase-activating protein 1 (IQGAP1) was confirmed as a novel target of HG, where HG appears to bind to the Glu-1019 site of IQGAP1 to exert its osteogenic effects. Data subsequently suggested that HG promoted phosphorylation of SMAD2\/3 and regulated the SMAD2\/3 pathway by inhibiting SMAD4 ubiquitination. Overall, the findings highlight HG as a new small-molecule drug to promote bone formation through SMAD2\/3 pathway in osteoporosis. \u00a9 2023 American Society for Bone and Mineral Research (ASBMR).<\/jats:p>\n                  <jats:p>Abstract<\/jats:p>\n                  <jats:p>Higenamine is elucidated to upregulate SMAD2\/3 signaling through binding IQGAP1, which leads to the inhibition of SMAD4 ubiquitination and thereby promotion of the expression of key osteogenesis genes.<\/jats:p>","DOI":"10.1002\/jbmr.4800","type":"journal-article","created":{"date-parts":[[2023,3,13]],"date-time":"2023-03-13T00:24:45Z","timestamp":1678667085000},"page":"775-791","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Higenamine Promotes Osteogenesis Via IQGAP1\/SMAD4 Signaling Pathway and Prevents Age- and Estrogen-Dependent Bone Loss in Mice"],"prefix":"10.1093","volume":"38","author":[{"given":"Hui","family":"Dong","sequence":"first","affiliation":[{"name":"Jinan Central Hospital Shandong University Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6897-0279","authenticated-orcid":false,"given":"Ronghan","family":"Liu","sequence":"additional","affiliation":[{"name":"Central Hospital Affiliated to Shandong First Medical University Shandong First Medical University & Shandong Academy of Medical Sciences Jinan China"},{"name":"Jinan Central Hospital Shandong University Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Zou","sequence":"additional","affiliation":[{"name":"Jinan Central Hospital Shandong University Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengxin","family":"Jin","sequence":"additional","affiliation":[{"name":"Central Hospital Affiliated to Shandong First Medical University Shandong First Medical University & Shandong Academy of Medical Sciences Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianning","family":"Kang","sequence":"additional","affiliation":[{"name":"Central Hospital Affiliated to Shandong First Medical University Shandong First Medical University & Shandong Academy of Medical Sciences Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Zhang","sequence":"additional","affiliation":[{"name":"Central Hospital Affiliated to Shandong First Medical University Shandong First Medical University & Shandong Academy of Medical Sciences Jinan China"},{"name":"Jinan Central Hospital Shandong University Jinan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Weifang Medical University Weifang 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