Abstract:Objective: To investigate the effects of icariin (ICA) on in vitro osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) and the phosphatidylinositol 3-kinase (PI3K)/serine threonine protein kinase (AKT) pathway. Methods: The third generation of rat BMSCs were randomly assigned into the control group (cultured in DMEM medium), dexamethasone (DXMS) group (cultured in DMEM medium supplemented with 10-6mol/L DXMS), L-ICA, M-ICA, H-ICA groups (DXMS induced DMEM medium+10-8, 10-7, 10-6mol/L ICA), and ICA+740Y-P group (DXMS induced DMEM medium+10-6mol/L ICA+30μmol/L PI3K activator 740Y-P). CCK8 method was used to examine the proliferation of rat BMSCs. Alizarin red staining was used to examine the mineralization ability of rat BMSCs induced by bone orientation (cultured in DMEM medium with a final concentration of 100nM DXMS+50μM vitamin+2 mM L-glutamine+10mM β-glycerophosphate). ELISA kit was used to examine the expression of ALP, ROS, and SOD in rat BMSCs. Western blot was used to examine the expression of PI3K/AKT pathway proteins in rat BMSCs. Results: The cytoplasmic staining in the DXMS group was relatively light, with lower A450 values, ALP, and SOD compared to the control group, while ROS, p-PI3K/PI3K, and p-AKT/AKT were higher than those in the control group (P<0.05); in the L-ICA group, M-ICA group, and H-ICA group, the cytoplasmic staining deepened, with higher A450 values, ALP, and SOD compared to the DXMS group, while ROS, p-PI3K/PI3K, and p-AKT/AKT were lower than those in the DXMS group (P<0.05); in the ICA + 740Y-P group, the cytoplasmic staining became lighter, with lower A450 values, ALP, and SOD compared to the H-ICA group, while ROS, p-PI3K/PI3K, and p-AKT/AKT were higher than those in the H-ICA group (P<0.05). Conclusion: ICA may inhibit the PI3K/AKT pathway and promote osteogenic differentiation of rat BMSCs.
季伟, 刘远航, 秦陈林, 欧宜林. 淫羊藿苷对大鼠骨髓间充质干细胞体外成骨分化及PI3K/AKT通路的影响[J]. 河北医学, 2025, 31(12): 2032-2037.
JI Wei, LIU Yuanhang, CAI Chenlin, et al. Effects of Icariin on in Vitro Osteogenic Differentiation and PI3K/AKT Pathway of Rat Bone Marrow Mesenchymal Stem Cells. HeBei Med, 2025, 31(12): 2032-2037.
[1] Tan B,Li W,Zeng P,et al.Epidemiological study based on China osteonecrosis of the femoral Head database[J].Orthop Surg,2021,13(1):153-160. [2] Li W,Li W,Zhang W,et al.Exogenous melatonin ameliorates steroid-induced osteonecrosis of the femoral head by modulating ferroptosis through GDF15-mediated signaling[J].Stem Cell Res Ther,2023,14(1):171-189. [3] Sun Y,Liang M,Xing Y,et al.Cyasterone has a protective effect on steroid-induced osteonecrosis of the femoral head[J].PLoS One,2023,18(10):3530-3544. [4] Yang X,Shi L,Li A,et al.Phase-contrast imaging with synchrotron hard X-ray reveals the effect of icariin on bone tissue morphology and microstructure in rabbits with early glucocorticoid-induced osteonecrosis of the femoral head[J].Front Cell Dev Biol,2023,11(5):532-539. [5] Li H,Zhang Y,Hao Y,et al.Proanthocyanidins inhibit osteoblast apoptosis via the PI3K/AKT/Bcl-xL pathway in the treatment of steroid-Induced osteonecrosis of the femoral head in rats[J].Nutrients,2023,15(8):1936-1949. [6] Chen Y,Pan X,Zhao J,et al.Icariin alleviates osteoarthritis through PI3K/Akt/mTOR/ULK1 signaling pathway[J].Eur J Med Res,2022,27(1):204-214. [7] 孔令驰,孙兴亮,左荣台,等.早期移植生长分化因子11表达沉默的骨髓基质干细胞促进大鼠激素性股骨头坏死区成骨的研究[J].中华创伤骨科杂志,2019,21(9):802-809 [8] Yue J,Yu H,Liu P,et al.Preliminary study of icariin indicating prevention of steroid-induced osteonecrosis of femoral head by regulating abnormal expression of miRNA-335 and protecting the functions of bone microvascular endothelial cells in rats[J].Gene,2021,766(1):145128. [9] Zhang X Y,Li H N,Chen F,et al.Icariin regulates miR-23a-3p-mediated osteogenic differentiation of BMSCs via BMP-2/Smad5/Runx2 and WNT/β-catenin pathways in osteonecrosis of the femoral head[J].Saudi Pharm,2021,29(12):1405-1415. [10] Wang D,Liu Y,Tang D,et al.Induction of PI3K/Akt-mediated apoptosis in osteoclasts is a key approach for buxue tongluo pills to treat osteonecrosis of the femoral head[J].Front Pharmacol,2021,12(1):729909-729918. [11] Wang X Y,Gong L J,Huang J M,et al.Pinocembrin alleviates glucocorticoid-induced apoptosis by activating autophagy via suppressing the PI3K/Akt/mTOR pathway in osteocytes[J].Eur Pharmacol,2020,880(1):173212. [12] Tang Y,Li Y,Xin D,et al.Icariin alleviates osteoarthritis by regulating autophagy of chondrocytes by mediating PI3K/AKT/mTOR signaling[J].Bioengineered,2021,12(1):2984-2999. [13] Li H,Ji L,Shen Y,et al.Bushen jianpi quyu formula alleviates myelosuppression of an immune-mediated aplastic anemia mouse model via inhibiting expression of the PI3K/AKT/NF-κB signaling pathway[J].Evid Based Complement Alternat Med,2022,1(1):297-306.