Tian Yawei, Liu Huiming, Bao Xiaoxue, Li Yukun
Department of Endocrinology, Hebei Medical University Third Hospital, Shijiazhuang, China.
Department of Stomatology, Hebei Medical University Second Hospital, Shijiazhuang, China.
Front Pharmacol. 2025 Mar 10;16:1539411. doi: 10.3389/fphar.2025.1539411. eCollection 2025.
Diabetes mellitus is a global disease in which alterations in the internal environment disrupt the bone-fat balance, contributing to osteoporosis. Semaglutide, a single-target, long-acting glucagon-like peptide-1 receptor agonist (GLP-1RA), has been shown to promote osteogenesis , but the underlying mechanism remains unclear. In this study, the ability of Semaglutide to promote the proliferation of bone-derived mesenchymal stem cells (BMSCs) was determined by CCK-8 kit and flow cytometry, Alkaline phosphatase (ALP) staining and alizarin red S staining showed that semaglutide increased ALP activity and the proportion of mineralised nodules during induction of osteogenesis, wound healing assay to evaluate the pro-migratory ability of semaglutide on BMSCs.Western blotting and RT-PCR showed that semaglutide promoted the mRNA and protein expression of osteocalcin (OCN) and Runt-related transcription factor 2 (RUNX2), and further determined the OCN expression level by immunofluorescence. RNA sequencing was performed to analyze the mechanisms underlying BMSC osteogenesis after semaglutide intervention. Enrichment of RNA sequencing data indicated that the Wnt/LRP5/β-catenin pathway was activated after treatment with semaglutide. Western blotting further confirmed the upregulation of Wnt pathway-associated protein levels by semaglutide. Dickkopf-1 (DKK1) and LiCl (lithium chloride) are common inhibitors and agonists of the Wnt/β-catenin pathway. The addition of semaglutide resulted in the partial reversal of the inhibitory effect of DKK1 on osteogenic differentiation, with the administration of LiCl and semaglutide further accelerating the osteogenic process. In addition to alterations in gene and protein expression levels, these changes are also reflected in alkaline phosphatase (ALP) activity and calcium deposition. Therefore, we suggest that semaglutide can promote the proliferation and osteogenic differentiation of BMSCs via the Wnt/LRP5/β-catenin signalling pathway.
糖尿病是一种全球性疾病，其体内环境的改变会破坏骨-脂肪平衡，导致骨质疏松。司美格鲁肽是一种单靶点、长效胰高血糖素样肽-1受体激动剂（GLP-1RA），已被证明可促进骨生成，但其潜在机制仍不清楚。在本研究中，通过CCK-8试剂盒和流式细胞术测定司美格鲁肽促进骨源性间充质干细胞（BMSC）增殖的能力，碱性磷酸酶（ALP）染色和茜素红S染色显示，司美格鲁肽在诱导成骨过程中增加了ALP活性和矿化结节的比例，通过伤口愈合试验评估司美格鲁肽对BMSC的促迁移能力。蛋白质印迹法和RT-PCR显示，司美格鲁肽促进了骨钙素（OCN）和Runt相关转录因子2（RUNX2）的mRNA和蛋白表达，并通过免疫荧光进一步测定了OCN表达水平。进行RNA测序以分析司美格鲁肽干预后BMSC成骨的潜在机制。RNA测序数据的富集表明，司美格鲁肽处理后Wnt/LRP5/β-连环蛋白通路被激活。蛋白质印迹法进一步证实司美格鲁肽上调了Wnt通路相关蛋白水平。Dickkopf-1（DKK1）和氯化锂（LiCl）是Wnt/β-连环蛋白通路常见的抑制剂和激动剂。添加司美格鲁肽导致DKK1对成骨分化抑制作用的部分逆转，而给予LiCl和司美格鲁肽进一步加速了成骨过程。除了基因和蛋白表达水平的改变外，这些变化还反映在碱性磷酸酶（ALP）活性和钙沉积上。因此，我们认为司美格鲁肽可通过Wnt/LRP5/β-连环蛋白信号通路促进BMSC的增殖和成骨分化。
