前列腺素E2诱导大鼠骨髓间充质干细胞成骨分化过程中p38MAPK信号通路蛋白的表达

Expressions of p38MAPK signaling pathway proteins during osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by prostaglandin E2

目的:探讨前列腺素E2对大鼠骨髓间充质干细胞(BMSCs)成骨分化的影响及其机制。方法:将第3代大鼠BMSCs分为3组,空白组用完全培养基培养,前列腺素E2组用含1×10-5 g/L前列腺素E2的培养液培养,前列腺素E2+抑制剂组用p38MAPK阻断剂SB203580处理30 min后用含1×10^-5 g/L前列腺素E2的培养液培养。干预14 d后,Western blot法检测细胞中p38MAPK和磷酸化p38MAPK(p-p38MAPK)蛋白的表达,检测细胞中碱性磷酸酶(ALP)活性,qRT-PCR法检测细胞中ALP、RunX2、骨桥蛋白(OPN)、骨形态发生蛋白2(BMP-2)和核心结合因子α1(Cbfα1)mRNA的表达。结果:与空白组相比,前列腺素E2组细胞中p-p38MAPK/p38MAPK比值、ALP活性,以及ALP、Runx2、OPN、BMP-2、Cbfα1 mRNA的表达水平均升高(P<0.05);与前列腺E2组比较,前列腺素E2+SB203580组细胞中上述各指标降低(P<0.05)。结论:前列腺素E2可通过激活p38MAPK信号通路诱导大鼠BMSCs成骨分化。

Aim:To investigate the effects of prostaglandin E2 on osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs)and its mechanism.Method:The 3rd generation rat BMSCs were divided into 3 groups.BMSCs of the blank group were cultured by complete medium culture,BMSCs of prostaglandin E2 group were cultured by 1×10^-5 g/L prostaglandin E2,and those of prostaglandin E2+SB203580 group were treated by p38MAPK blocker SB203580 for 30 min and then treated by 1×10^-5 g/L prostaglandin E2.After 14 days of intervention,the expressions of p38MAPK and p-p38MAPK proteins were detected by Western blot,the activity of ALP was tested,and the expressions of ALP,OPN,Runx2,BMP-2 and Cbfα1 mRNA were examed by qRT-PCR.Results:Compared with those of the blank group,the ratio of p-p38MAPK/p38MAPK,ALP activity,and the expressions of ALP,Runx2,OPN,BMP-2 and Cbfα1 mRNA in BMSCs of the prostaglandin E2 group were significantly higher(P<0.05),while the indexes mentioned above in the prostaglandin E2+SB203580 group were significantly lower than those in the prostaglandin E2 group(P<0.05).Conclusion:Prostaglandin E2 can induce osteogenic differentiation of rat BMSCs by activating p38MAPK signaling pathway.