ERK信号通路在檞皮素促大鼠MSCs成骨分化中的作用  

The roles of ERK signaling pathway in osteogenic differentiation of rat MSCs promoted by quercetin

作　　者：奉水旺[1] 杨丽[1] 王攀攀[2] 李淑琴[1] 汪甜[1] 尹素娟[1] 张荣华[1] 

机构地区：[1]暨南大学药学院中药学教研室,广东广州510632 [2]暨南大学医学院,广东广州510632

出　　处：《中国病理生理杂志》2012年第8期1477-1481,共5页Chinese Journal of Pathophysiology

基　　金：广东省教育厅育苗工程(No.34310016)

摘　　要：目的:观察细胞外信号调节激酶(ERK)信号通路在檞皮素(QUE)促进SD大鼠骨髓间充质干细胞(MSCs)成骨分化过程中的作用。方法:(1)用0.01μmol/L、0.1μmol/L、1μmol/L、10μmol/L和100μmol/L QUE干预MSCs,MTT法检测各浓度QUE对MSCs增殖的影响,碱性磷酸酶(ALP)测定试剂盒检测各浓度QUE对MSCsALP表达的影响;(2)用ERK1/2抑制剂干预后,加入QUE,用ALP测定试剂盒检测ALP的表达,ELISA法检测Ⅰ型胶原(ColⅠ)和骨钙素(BGP)的表达,Western blotting检测ERK1/2的表达,荧光定量PCR检测转化生长因子β1(TGF-β1)mRNA、骨形成蛋白2(BMP-2)mRNA和核心结合因子α1(Cbfα1)mRNA表达。结果:(1)0.1μmol/L、1μmol/L和10μmol/L QUE剂量依赖性地促进MSCs ALP的表达,同时能促进MSCs的增殖;(2)与空白组相比,QUE组ALP、BGP和ColⅠ表达均增加(P<0.01),加入ERK1/2抑制剂后,磷酸化的ERK1/2表达减少(P<0.05),同时ALP、BGP和ColⅠ表达降低(P<0.01);(3)与空白组比较,QUE组TGF-β1mRNA、BMP-2 mRNA和Cbfα1mRNA的表达均增加(P<0.05),加入ERK1/2抑制剂后这3个基因的表达都下降(P<0.05)。结论:一定浓度的QUE能促进MSCs的增殖和成骨分化,ERK通路的激活在此过程中起到了重要的作用。To study the roles of extracellular signal - regulated kinase （ERK） signal pathway in the process of osteogenic differentiation in rat mesenchymal stem cells （MSCs） promoted by quercetin （QUE）. METHODS： The optimal concentration of QUE for promoting osteogenic differentiation of rat MSCs was determined by MTY and alkaline pbosphatase （ALP） detection. The activity of ALP was detected by the ALP detection kit. The expression of bone Gla pro- tein （BGP） and collagen type I （Col I ） was observed by ELISA analysis. MSCs were exposed to QUE at optimal concen- tration with or without ERK1/2 inhibitor PD98059. Non - phosphorylated and phosphorylated expression of ERK1/2 was analyzed by Western blotting. The mRNA expression of transforming growth factor 131 （ TGF-β1）, bone morphogenetic protein 2 （ BMP - 2） and core binding factor al （ Cbfotl ） was measured by fluorescence quantitative PCR. RESULTS ： QUE at concentrations of 0. 1 Iμmol/L, 1 Ixmol/L and 10 pμmol/L induced the expression of ALP in MSCs in a dose- de- pendent manner, and also promoted MSCs proliferation. The expression levels of ALP, BGP and Col I were higher in QUE group, and was lower in PD89059 group than those in control group. Compared with control group, the level of phosphoryl- ated ERK1/2, and the mRNA expression of TGF- 131, BMP-2 and Cbfotl increased in QUE group. The mRNA expres- sion of TGF - β1, BMP - 2 and Cbfal in QUE ＋ PD98059 group decreased as compared with QUE group. CONCLU- SION： QUE promotes osteogenic differentiation of MSCs by activating ERK signaling pathway.

关 键 词：骨髓间充质干细胞 成骨分化 有丝分裂原活化蛋白激酶类 信号通路 

分 类 号：R363[医药卫生—病理学]