为了研究淫羊藿苷(ICA)抑制Notch信号通路对骨髓间充质干细胞(BMSCs)成骨分化.选取8周龄SPF级健康雌性大鼠40只,手术摘除双侧卵巢建立大鼠骨质疏松症模型.收集BMSCs细胞进行体外分离培养,取传代第三代细胞进行鉴定,试验组加入浓度为0.8 g/100 mL ICA,对照组加入同剂量的雌激素,对细胞进行培养.收集培育的细胞上清液,检测碱性磷酸酶(ALP)和骨钙素(OCN)水平及Notch信号通路中相关蛋白Notch1、Jagged-1、CBF1的表达水平.结果表明,流式细胞检测P3代骨髓间充质干细胞表面抗原,CD90、CD44、CD34、CD11阳性率分别为98.2％,93.3％,2.6％,4.4％,确认细胞鉴定为骨髓干细胞.与对照组比较,试验组ALP和OCN活性明显升高、另外Noth1、CBF1、Jagged-1蛋白表达量均较对照组降低.淫羊藿苷通过上调ALP和OCN水平及抑制Notch通路中Notch1、CBF1、Jagged-1蛋白的表达,实现对去卵巢骨质疏松症大鼠BMSCs的增殖和成骨细胞分化.
A Study on Icariin Inhibiting Notch Signaling Pathway and Promoting Differentiation of Bone Marrow Mesenchymal Stem Cells
To study the effect of icariin(ICA)on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)by inhibiting Notch signaling pathway,a total of 40 8-week-old SPF healthy female rats were selected and bilateral ovaries were surgically removed to establish an osteoporosis model.BMSCs were isolated and cultured in vitro,and the third generation cells were identified.The experimental group was treated with 0.8 g/100 mL ICA.The supernatant of cultured cells was collected to detect the levels of alkaline phosphatase(ALP),osteocalcin(OCN)and Notch signaling pathway-related proteins Notch1,Jagged-1 and CBF1.The results showed that the positive rates of CD90,CD44,CD34 and CD11 were 98.2％,93.3％,2.6％and 4.4％,respectively,and the cells were identified as bone marrow stem cells.Compared with the control group,the ALP and OCN activity of the experimental group was significantly increased,and the protein expression of Noth1,CBF1 and Jagged-1 was decreased.Icariin can promote the proliferation and osteogenic differentiation of BMSCs in ovariectomized osteoporosis rats by upregulating the levels of ALP and OCN and inhibiting the expression of Notch1,CBF1,and Jagged-1 proteins in the Notch pathway.
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