摘要
目的:运用血清药理学方法研究滋肾降糖丸对小鼠骨髓基质干细胞(MSC)模型成骨分化的作用及其机制。方法:首先空腹6h后的小鼠口服滋肾降糖溶液,剂量为1.5g·kg-1,随后在0、0.5、1、2和4h不同时间段收集含药血清;随后用茜素红染色和碱性磷酸酶活性检测的方法调查上述含药血清对MSC成骨诱导分化的影响;最后用实时荧光定量PCR的方法进一步调查含药血清对MSC成骨诱导过程中相关关键基因骨形态发生蛋白(BMP2)及成骨特异性转录因子(RunX2)的表达。结果:含药血清可显著促进MSC的成骨分化,以2h后收集的血清效果最强。Q-RT-PCR实验结果显示,2h含药血清显著促进诱导后第3、6dMSC细胞BMP2及RunX2基因的表达。结论:滋肾降糖丸抗骨质疏松的作用可能与上调MSC上BMP2及RunX2的基因表达,从而促进MSC的成骨分化有关。
OBJECTIVE:To investigate the effect of Zishen Jiantang pills(kidney-nourishing hypoglycemic pills) on osteogenesis differentiation in mesenchymal stem cells(MSC) model in mice and the underlying mechanisms by using serum pharmacological method.METHODS:After fasting for 6 h,the mice were orally administrated with Zishen Jiantang pills at a dose of 1.5 g·kg ^-1 followed by sampling of drug-containing serum at 0,0.5,1,2,and 4 h,respectively.Then,the effects of drug-containing serum on osteogenesis were investigated by ARS(alizarin red) staining and APA(alkaline phosphatase activities) assaying;and the expressions of BMP2(key gene in osteogenesis) and RunX2 were detected by real-time RT-PCR method.RESULTS:The serum containing active components from Zishen Jiantang pills significantly promoted the osteogenesis of MSC,with the serum sample collected at 2 h showing the highest potency.After exposing to drug-containing serum collected at 2 h,the expressions of BMP2 and RunX2 were significantly increased at 3 and 6 days of induction of osteogenesis.CONCLUSION:The anti-osteoporosis effect of Zishen Jiangtang pills might be attributed to the promoted osteogenesis differentiation of MSC induced by the upregulated expressions of BMP2 and RunX2.
出处
《中国医院用药评价与分析》
2010年第10期905-907,共3页
Evaluation and Analysis of Drug-use in Hospitals of China