Objective To investigate the effects of puerarin on the osteogenic differentiation of bone marrow mesenchymal stem/stromal cells (BMSCs) and the mechanisms involved. Methods BMSCs were divided into a control group and treatment groups with different concentrations of puerarin. Cell proliferation was assessed using the CCK-8 assay. Alkaline phosphatase (ALP) and Alizarin Red staining were used to evaluate osteogenic differentiation. Seahorse technology was used to analyze cellular glycolytic function, while q-PCR detected the mRNA expression of miR-141-3p, Runx2, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), interleukin-10 (IL-10), transforming growth factor-β (TGF-β), hexokinase 2 (HK2), and lactate dehydrogenase A (LDHA). Western blot was employed to analyze the expression of Runx2 protein. The targeting relationship between miR-141-3p and Runx2 was determined through a dual-luciferase reporter gene experiment. Results Compared with the control group, puerarin concentration dependently promoted BMSCs proliferation (P<0.01), ALP activity (P<0.01) and calcium nodule deposition (P<0.01). Seahorse analysis showed that puerarin significantly enhanced the glycolytic function of BMSCs (P<0.01). Puerarin could also significantly down-regulated the expression of miR-141-3p (P<0.01), up-regulated the mRNA miR-141-3p (P<0.01) and the mRNA and protein levels of Runx2 (P<0.01). Meanwhile, puerarin promoted the expression of NGF, BDNF, IL-10 and TGF-β (P<0.01). Double luciferase reporter gene assay showed that miR-141-3p could directly target Runx2 3'UTR. Conclusion Puerarin alleviates the inhibition of Runx2 by downregulating miR-141-3p, promotes osteogenic differentiation, and exerts multidimensional bone promoting effects by activating neurotrophic factor signaling and improving the bone immune microenvironment.
Key words
puerarin /
osteoblasts /
microRNA-141-3p /
neurotrophic factor
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References
[1] Li CJ, Xiao Y, Sun YC, et al. Senescent immune cells release grancalcin to promote skeletal aging[J]. Cell Metab,2022,34:184–185.
[2] 刘畅,李静,江思婧,等.microRNAs调节骨免疫微环境的研究进展[J].实用医院临床杂志,2022,19:214–217.
[3] Chen MH, Wang JX, Yang YH, et al. The interplay of estrogen, gut microbiome, and bone immunity in osteoporosis[J]. Cell Commun Signal,2025,23:516.
[4] Zhuang PZ, Chen Y, Zhang Y, et al.Regulating macrophage glucose metabolism homeostasis via mitochondrial rheostats by short fiber-microsphere scaffolds for bone repair[J]. Bioact Mater,2025,49:399–417.
[5] Xiang YL, Li ZL, He X, et al. Puerarin relives inflammation, bone destruction and facilitates osteogenic differentiation in periodontitis by enhancing mitochondrial autophagy via activating mitochondrial Mitofusin 2[J]. Stem Cell Res Ther, 2025, 16:218.
[6] 周凡,高扬,胡艳平,等.葛根素调控SIRT1-FOXO1通路抑制成骨细胞凋亡[J].中国骨质疏松杂志,2025,31:671–675,702.
[7] 魏振朴,张文明,孙攀,等.miR-141及成骨基因Dlx5、Msx2、Runx2在去卵巢骨质疏松模型小鼠BMSCs成骨分化中的表达[J].康复学报,2019,29:37–43.
[8] 张亚军,陈金洪,王勇,等.葛根素通过细胞外信号调节激酶1/2和p38丝裂原活化蛋白激酶信号通路刺激成骨分化和骨形成的机制[J].中华实验外科杂志,2019,36(5):973-973.
[9] 温乐,王凌伟,王婷,等.葛根素对骨质疏松大鼠的经典Wnt信号通路调控的影响[J].神经药理学报,2025,15:1–6.
[10] Li X, Lin XQ, Chen DD, et al. B-cell lymphoma-2 phosphorylation at Ser70 site-related autophagy mediates puerarin-inhibited the apoptosis of MC3T3-E1 cells during osteoblastogenesis[J]. J Tradit Chin Med, 2024, 44:27–34.
[11] 支旺,温惠慧,崔志强.葛根素通过抑制MAPK/NF-κB信号通路调节LPS诱导的人牙髓干细胞的炎症损伤和成骨分化[J].实用口腔医学杂志,2023,39:648–654.
[12] Katsianou AM, Gargalionis NA, Papavassiliou AK, et al. The critical role of transcription factor RUNX2 in bone mechanobiology[J]. Cells,2025,15:50.
[13] Itting TP, Kruppke B, Hanke T, et al. BDNF improves the survival of mesenchymal stem cells cultured on pre-structured gelatin material containing strontium and calcium phosphates for bone regeneration[J]. Front Bioeng Biotechnol,2025,13:1596846.
[14] Leena S, Asha B, Kumar PM, et al. Regulatory B cells (Bregs) inhibit osteoclastogenesis and play a potential role in ameliorating ovariectomy-induced bone loss[J]. Front Immunol,2021,12:691081.
[15] Kitazawa R, Haraguchi R, Kitazawa S.A variant form of receptor activator of nuclear factor-κB functions as an osteoprotegerin mimic in bone and cardiac development[J]. Sci Rep,2025,15:27919.
[16] Bertels CJ, He G, Long F.Metabolic reprogramming in skeletal cell differentiation[J]. Bone Res,2024,12:57.
[17] Antonia D, PaulRichard K, Shan J, et al. Glucose metabolism in osteoblasts in healthy and pathophysio-logical conditions[J]. Int J Mol Sci,2021,22:4120.
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