[1]季伟,刘远航,张倩,等.淫羊藿苷对激素性股骨头坏死人骨髓间充质干细胞成骨分化的影响[J].中国中医骨伤科杂志,2025,33(06):14-20.[doi:10.20085/j.cnki.issn1005-0205.250603]
　JI Wei,LIU Yuanhang,ZHANG Qian,et al.Effect of Icariin on Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells with Hormone-Induced Femur Head Necrosis[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2025,33(06):14-20.[doi:10.20085/j.cnki.issn1005-0205.250603]
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淫羊藿苷对激素性股骨头坏死人骨髓间充质干细胞成骨分化的影响()

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第33卷
期数:
2025年06期
页码:
14-20
栏目:
实验研究
出版日期:
2025-06-15

文章信息/Info

Title:
Effect of Icariin on Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells with Hormone-Induced Femur Head Necrosis
文章编号:
1005-0205(2025)06-0014-07
作者:
季伟1; 刘远航1; 张倩1; 欧宜林2△
1武汉市第三医院(武汉大学同仁医院)(武汉,430060) 2钟祥市中医院
Author(s):
JI Wei1; LIU Yuanhang1; ZHANG Qian1; OU Yilin2△
1Wuhan Third Hospital(Tongren Hospital of Wuhan University),Wuhan 430060,China; 2Zhongxiang Traditional Chinese Medicine Hospital,Zhongxiang 431900,Hubei China.
关键词:
淫羊藿苷;  激素性股骨头坏死;  人骨髓间充质干细胞;  成骨、成脂分化;  磷脂酰肌醇3-激酶/丝氨酸-苏氨酸激酶通路
Keywords:
icariin;  steroid-induced avascular necrosis of the femoral head;  human bone marrow mesenchymal stem cells;  osteogenic and adipogenic differentiation;  phosphatidylinositol 3-kinase/serine threonine kinase pathway
分类号:
R681.8
DOI:
10.20085/j.cnki.issn1005-0205.250603
文献标志码:
A
摘要:
目的:基于磷脂酰肌醇3-激酶/丝氨酸-苏氨酸激酶(PI3K/AKT)通路探讨淫羊藿苷对激素性股骨头坏死人骨髓间充质干细胞(hBMSCs)成骨分化能力的影响。方法:体外培养人骨髓间充质干细胞,通过成骨、成脂诱导分化进一步鉴定人骨髓间充质干细胞的分化能力。以地塞米松诱导人骨髓间充质干细胞模型,以0,0.1,0.5,1,5,10,50,100 nmol/L的淫羊藿苷干预24 h后,CCK-8实验筛选出合适的淫羊藿苷浓度进行后续研究(CCK-8法)。将人骨髓间充质干细胞分为对照组、模型组、PI3K激活剂组(10 μmol/L 740YP组)、淫羊藿苷组、淫羊藿苷+PI3K抑制剂组(淫羊藿苷+10 mmol/L LY294002组)。CCK-8法检测各组人骨髓间充质干细胞活力; 染色法检测人骨髓间充质干细胞的成骨、成脂分化能力; RT-qPCR法检测成骨分化相关基因表达水平; 比色法检测碱性磷酸酶(ALP)活性; Western Blot法检测各组人骨髓间充质干细胞中PI3K/AKT通路有关蛋白表达量。结果:第3代人骨髓间充质干细胞生长良好,成骨、成脂诱导分别显示有钙化结节、红色脂滴出现,说明人骨髓间充质干细胞具有成骨和成脂分化的能力。CCK-8法筛选淫羊藿苷的浓度为1 nmol/L用于后续实验。与对照组相比,模型组钙化结节数量、ALP活性、成骨因子(ALP、RUNX2、OPN、BMP2)的基因表达水平以及p-PI3K/PI3K、p-AKT/AKT和Bcl-2蛋白表达水平明显降低,脂滴数量、成脂因子(PPARγ、C/EBPα)的基因表达水平以及Bax蛋白表达水平显著升高,差异有统计学意义(P<0.05); 与模型组相比,PI3K激活剂组和淫羊藿苷组钙化结节数量、ALP活性、成骨因子的基因表达水平以及p-PI3K/PI3K、p-AKT/AKT和Bcl-2蛋白表达水平显著升高,脂滴数量、成脂因子的基因表达水平以及Bax蛋白表达水平明显下降,差异有统计学意义(P<0.05); 与淫羊藿苷组相比,淫羊藿苷+PI3K抑制剂组钙化结节数量、ALP活性、成骨因子的基因表达水平以及p-PI3K/PI3K、p-AKT/AKT和Bcl-2蛋白表达水平明显降低,脂滴数量、成脂因子的基因表达水平以及Bax蛋白表达水平显著升高,差异有统计学意义(P<0.05)。结论:淫羊藿苷可能通过激活PI3K/AKT通路,进而增强地塞米松诱导的人骨髓间充质干细胞成骨分化能力和细胞活性。
Abstract:
Objective:To investigate the effect of icariin on osteogenic differentiation ability of human bone marrow mesenchymal stem cells(hBMSCs)with femoral head based on the phosphatidylinositol 3-kinase/serine threonine kinase(PI3K/AKT)pathway.Methods:hBMSCs were cultured in vitro and their differentiation ability was further identified by inducing osteogenic and adipogenic differentiation.Dexamethasone induced hBMSCs model,and after 24 h of intervention with 0,0.1,0.5,1,5,10,50,and 100 nmol/L icariin,the appropriate icariin concentration was screened by CCK-8 experiment for subsequent research(CCK-8 method).hBMSCs were assigned into control group,model group,PI3K activator group(10 μmol/L 740YP group),icariin group,and icariin+PI3K inhibitor group(icariin+10 mmol/L LY294002 group).CCK-8 was used to detect the viability of hBMSCs in each group.Alizarin red staining and oil red O staining were used to detect the differentiation abilities of hBMSCs.RT-qPCR method was used to detect the expression levels of osteogenic differentiation related genes.Colorimetric method was used to detect alkaline phosphatase(ALP)activity.Western Blot method for detecting protein expression levels related to the PI3K/AKT pathway.Results:The 3rd generation hBMSCs grew well,with calcified nodules and red lipid droplets appearing during osteogenic and adipogenic induction,indicating that hBMSCs had osteogenic and adipogenic differentiation abilities.The concentration of icariin screened by CCK-8 was 1 nmol/L for subsequent experiments.Compared with the control group,the number of calcified nodules,ALP activity,gene expression levels of osteogenic factors(ALP,RUNX2,OPN,BMP2),protein expression levels of p-PI3K/PI3K,p-AKT/AKT,and Bcl-2 were prominently lower in the model group,while the number of lipid droplets,gene expression levels of adipogenic factors(PPARγ,C/EBPα),and protein expression level of Bax were obviously higher(P<0.05).Compared with the model group,the number of calcified nodules,ALP activity,gene expression levels of osteogenic factors,protein expression levels of p-PI3K/PI3K,p-AKT/AKT,and Bcl-2 were obviously higher in the PI3K activator group and icariin group,while the number of lipid droplets,gene expression levels of adipogenic factors,and protein expression level of Bax were prominently lower(P<0.05).Compared with the icariin group,the number of calcified nodules,ALP activity,gene expression levels of osteogenic factors,protein expression levels of p-PI3K/PI3K,p-AKT/AKT,and Bcl-2 were prominently lower in the icariin+PI3K inhibitor group,while the number of lipid droplets,gene expression levels of adipogenic factors,and protein expression level of Bax were obviously higher(P<0.05).Conclusion:Icariin can enhance dexamethasone induced osteogenic differentiation and cellular activity of hBMSCs by activating the PI3K/AKT pathway.

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备注/Memo

备注/Memo:
基金项目:武汉市卫健委中医药科研项目(WZ22Q14)
△通信作者 E-mail:dtvlzf@163.com
更新日期/Last Update: 2025-06-15