摘要
目的探讨麦角甾苷对大鼠骨髓基质细胞增殖和成骨分化的影响及机制。方法密度梯度离心法分离培养大鼠骨髓基质细胞并鉴定，细胞随机分为4组，分别以终浓度为100μmol／L（A组）、50μmol／L（B组）、25μmol／L（C组）和0μmol／L（D组）的麦角甾苷干预培养。24h、48h及72h后采用CCK-8法检测细胞增殖情况。于干预培养24h、48h和96h后进行碱性磷酸酶活性检测；72h后进行碱性磷酸酶染色。Westernblot及Realtime—PCR分别检测成骨相关蛋白及基因表达情况。结果加药干预24h和48h后，麦角甾苷对骨髓基质细胞增殖情况并无显著影响（P〉0．05）；加药72h后，A组吸光度值降低（P〈O．05）。A～C三组碱性磷酸酶表达量均较D组有所增高（P〈0．05），B组及C组较明显。Western Blot及Realtime—PCR检测结果显示A组、B组及C组BMP2、Smad1、Smad5、Smad8、Runx2、Osterix成骨相关蛋白及对应基因表达水平均高于D组（P〈0．05），而各加药组间差异不明显（P〉0．05）。结论麦角甾苷对体外培养的大鼠骨髓基质细胞增殖并无显著影响，并可以通过BMP2-Smads—Runx2-Osterix通路促进BMSCs成骨分化。
Objective To study the Effects of verhascoside on the proliferation and osteogenic differentiation of rat bone marrow stromal cells and its mechanism. Methods BMSCs were obtained and identified from rat bone marrow by density gradient centrifugation. BMSCs were divided into four groups and treated with verbaseoside at concentrations of 100 μmol/L（group A） ,50μmol/L（group B） ,25μmol/L（group C）,and 0μmol/L（group D）. The CCK-8 method was used to evaluate the BMSCs proliferation at 24h,48h and 72h after administration of the medication. The alkaline phosphatase activities were detected at 24h,48h and 96h after medication; Alkaline phosphatase staining was performed after 72h of medication. Western blot and Realtime-PCR were used to detect osteogenic related proteins and genes expression. Results The 24h and 48h drug interventions bad no obvious influence on the BMSCs proliferation（P〉 0.05）. After 72h of medication,group A decreased（P〈0.05）. The alkaline phosphatase expression level was higher in groups of A,B and C than that in group D,especially in the groups of B and D. Western blot and Realtime-PCR showed that the osteogenic related pro reins of gMP2,Smadl,Smad5,Smad8,Runx2,Osterix and genes expression level in groups of A B and C were higher than those of group D（P〈0.05）. With no significant changes in the three groups with different concentrations of verbascoside. Conclusion The verbascoside at different concentrations had no obvious effect on BMSCs proliferation, and can promote osteogenic differentiation of BMSCs through gMP2-Smads-Runx2-Osterix pathway.
出处
《检验医学》
CAS
2016年第B09期63-66,共4页
Laboratory Medicine
基金
国家自然科学基金（31170930）
关键词
麦角甾苷
骨髓基质细胞
增殖
成骨分化
verbaseoside
gone marrow stromal cells
proliferation
osteogenic differentiation
