松果菊苷含药血清诱导骨髓间充质干细胞成骨分化及BMP2表达的研究  

Effects of echinacoside serum on the proliferation and the osteogenic differentiation and expression of BMP2 in rat bone marrow mesenchymal stem cells

作　　者：田原[1,2] 邸阳[2] 包翠芬[2] 林宇涵[2] 秦书俭[2] 

机构地区：[1]辽宁中医药大学中西医结合基础专业,沈阳110847 [2]辽宁医学院基础医学院,锦州121001

出　　处：《中药药理与临床》2015年第4期60-64,共5页Pharmacology and Clinics of Chinese Materia Medica

摘　　要：目的:探讨松果菊苷含药血清促进大鼠骨髓间充质干细胞向成骨细胞分化的作用及可能的机制。方法:大鼠用松果菊苷(16mg/kg)灌胃4天后,取血清备用。采用全骨髓贴壁法培养大鼠骨髓间充质干细胞,取第3代细胞分为8组:空白对照组、松果菊苷含药血清组(2.5%、5%、7.5%、10%、15%、20%)、经典成骨诱导组(10%FBS+10 mmol/Lβ-甘油磷酸钠+10-8mol/L地塞米松+50μmol/L维生素C)。ELISA测定碱性磷酸酶(ALP)和骨钙素(BGP)的表达;免疫荧光细胞化学染色和Western blot检测BMP2蛋白的表达,RT-PCR检测BMP2mRNA的表达。结果:与经典成骨组相比,10%松果菊苷含药血清使BMSCs表达ALP(80.16U/L)和BGP(150.79 U/L)增强,BMP2免疫荧光、BMP2蛋白表达及BMP2mRNA的表达也明显增强。结论:松果菊苷含药血清可以诱导体外培养的骨髓间充质干细胞向成骨细胞分化,10%松果菊苷含药血清组的效果最佳,其作用机制与BMP2表达增加有关。Objective： To investigate the effects and mechanism of echinacoside serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Methods： Rats were given by gavage for 4 days with echinacoside（ 16 mg / kg）,then take them serum to prepare the drug-contained serum. Rat bone marrow mesenchymal stem cells were isolated by the whole bone marrow adherence method. Passage 3 cells were divided into 8 groups： The blank group is the L-DMEM which contain 5% fetal bovine serum and 20% blank rat serum; The concentrations of echinacoside serum groups（ 2. 5%,5%,7. 5%,10%,15%,20%）. the classic osteogenic induced group（ containing 10% fetal bovine serum,10 mmol / L β-glycerine sodium,10-8mol / L dexamethasone,and 50 μmol / L vitamin C）; The expression of ALP and BGP were detected by ELISA. Immunofluorescence was used to detect BMP2. The expression of BMP2 protein were detected by Western blot technique.Rt-pcr technique was used to detect the expression of BMP2 mRNA. Results： Expression of the ALP（ 80. 16 U / L） 、BGP（ 150. 79 U / L） 、immunofluorescence,BMP2 protein and BMP2 mRNA which were in 10% echinacoside serum group were significantly higher than those in the classic osteogenic induced group. Conclusions： Echinacoside serum could significantly promote the differentiation of the bone marrow mesenchymal stem cells,the effect of 10% echinacoside serum group was the best. The mechanism is associated with an enhancing expression of BMP2.

关 键 词：松果菊苷 含药血清 骨髓间充质干细胞 成骨分化 BMP2 

分 类 号：R285.5[医药卫生—中药学]