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目的 研究淫羊藿苷对脐带间充质干细胞的生物学作用,以及诱导其向成骨细胞分化.方法体外分离、培养及扩增人脐带间充质干细胞,免疫组化技术检测人脐带间充质干细胞(huCMSCs)表面特异标志物表达.取第3代细胞,设立2组,空白对照组加入基础培养基,实验组加入淫羊藿苷诱导.用倒置光学显微镜、碱性磷酸酶染色、碱性磷酸酶活性测定细胞.进行对比,研究人脐带间充质干细胞增殖和分化规律.结果 免疫组化结果显示huCMSCs表面标记CD44、CD34阴性,检测结果符合huCMSCs的特征.随着培养天数的增加,淫羊藿苷组诱导后细胞液中成骨细胞特征性碱性磷酸酶强阳性表达及钙沉积.淫羊藿苷组细胞内ALP含量与对照组比较差异有统计学意义(P<0.05).结论 huCMSCs可以在体外采用组织块贴壁培养法培养成功,淫羊藿苷对人脐带间充质干细胞的增殖和成骨分化具有促进作用,可作为骨性诱导因子.Abstract: Objective To study the biological effect of Icariine on human umbilical cord derived mesenchymal stem cells(huCMSCs) and inducing them to differentiate into osteoblasts. Methods MSCs were isolated and expanded in vitro and their surface antigens of huCMSCs were detected by immunohistochemistry. HuCMSCs were assigned into two groups. In the blank control group, huMSCs were incubated in basic medium. In the experimental group huMSCs were incubated in Icariine medium, The proliferation and differentiation of huMSCs were examined by inverted microscope, Alkaline phosphatase (ALP) staining and the determination of ALP activities. Results HuCMSCs were strongly positive for CD44, and negative for CD34; strongly positive expression for ALP and calcium deposition was detected from the second day to the fourth day in Icariine group; blank control group and Icariine group had significant differences (P < 0.05).Conclusion HuCMSCs can be successfully cultured from the adherent tissue pieces, Icariine enhances proliferation and osteogenic differentiation of huCMSCs and be used as an osteoinductive factor. 相似文献
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目的:建立房陵黄酒保健药酒中主要成分淫羊藿含量测定的方法。方法:采用高效液相色谱法对房陵黄酒保健药酒中的主药淫羊藿测定淫羊藿苷含量,采用Diamonsil^TM(钻石)C18(250mm×4.6mm,5μm),水-乙(70:30)为流动相,检测波长270nm,柱温30℃,流速1.0ml·min^-1。结果:淫羊藿苷在0.194~1.940μg范围内呈良好的线性关系,r=0.9999;平均回收率为98.22%(RSD=1.93%,n=6)。结论:所建立的方法快速准确、重复性好、简便易行,可作为房陵黄酒保健药酒的质量控制方法。 相似文献
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反相高效液相色谱法测定肾炎汤中淫羊藿苷的含量 总被引:2,自引:1,他引:2
目的:反相高效液相色谱法(RP-HPLC)法测定肾炎汤中淫羊藿苷的含量。方法:Hypersil C18柱,流动相为乙腈.水(27:73),在270nm波长处检测。结果:淫羊藿苷的线性范围为0.07-0.63μg,r=0.9999,平均加样回收率为99%。结论:方法简便,精密度和稳定性好,适用于测定肾炎汤中淫羊藿苷的含量。 相似文献
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目的:建立抗骨增生片中淫羊藿的高效液相色谱(HPLC)含量测定方法.方法:栗Agilent 1100高效液相色谱仪;流动相:乙腈-水(30:70);流速:1.0ml/min;捡测波长为270mn.结果:淫羊藿苷在0.01702mg~1.702mg范围内呈良好线性关系(r=0.9999),平均回收率为100.41%,RSD为1.91%(n=9).结论:该法操作简便,结果准确、专属性强. 相似文献
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高效液相色谱法测定强阳保肾丸中淫羊藿苷的含量 总被引:4,自引:0,他引:4
目的建立测定强阳保肾丸中淫羊藿苷含量的高效液相色谱法。方法强阳保肾丸经70%乙醇提取后,以聚酰胺柱纯化,采用高效液相色谱法测定其中淫羊藿苷的含量[固定相:大连依利特C18色谱柱(4.6 mm×250 mm,5 μm);流动相:甲醇 纯化水 冰醋酸(60∶39∶1,V/V);流速:1.0 mL·min-1;检测波长:270 nm]。结果淫羊藿苷在进样量为0.058~1.153 μg范围内线性关系良好(r=0.999 8),平均回收率为98.9 %,RSD=2.3%。结论该法操作简便,结果准确,可作为强阳保肾丸的质量控制方法。 相似文献
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