洛伐他汀对BMSCs成骨分化不同阶段Cbfa1、BMP-2表达的影响

EFFECT OF LOVASTATIN ON EXPRESSION LEVELS OF CBFA1 AND BMP-2 IN BMSCS AT DIFFERENT PHASE OF OSTEOGENIC DIFFERENTIATION

目的研究洛伐他汀促进大鼠骨髓基质干细胞(bone marrow stromal cells,BMSCs)向成骨细胞分化过程中特异性基因Cbfa1、BMP-2 mRNA表达的影响。方法取6周雌性SD大鼠的双侧股骨和胫骨骨髓细胞进行体外成骨诱导培养,于第一次传代后加入浓度为5μmol/L洛伐他汀或等量溶剂培养,分别于细胞培养第十四、二十一、二十八天提取RNA,并采用realtime RT-PCR法于上述时间点检测Cbfa1、BMP-2mRNA的表达,细胞培养至35d,采用茜素红染色观察细胞外基质矿化能力。结果①洛伐他汀可促进成骨诱导分化大鼠BMSCs细胞外基质的矿化能力。②mRNA表达水平:第十四天干预组Cbfa1的表达显著高于对照组,BMP-2的表达水平无显著组间差别;第二十一天干预组Cbfa1、BMP-2的表达均显著高于对照组;第二十八天BMP-2表达显著高于对照组,Cbfa1的表达无组间差别(P<0.05)。结论洛伐他汀可促进大鼠BMSCs向成骨细胞分化,其作用机制可能与在其分化不同阶段调控Cbfa1、BMP-2的表达有关。

Objective To investigate the effect of lovastatin on mRNA expression levels of Cbfal and BMP -2 in bone marrow stromal sells（BMSCs） at different phase of osteogenic differentiation. Methods BMSCs derived from the tibia and femur of 6 weeks old famale SD rats were cultured in vitro in osteoblastic- induced medium since the first subculture and divided into lovastatin stimulated group and control group, treated with 5umol/L lovastatin or vehicle as control repectively. Alizarin red staining was performed at 35th day to assess the mineralization of extracellular matrix. Real time RT - PCR was performed at 14th, 21th, 28th day to evaluate the expression levesls of Cbfal and BMP- 2 separately. Results Lovastatin could promote the extracellular matrix mineralization, mRNA expression： At 14th day, the expression of Cbfal was increased significantly in lovastatin stimulated group compared with control group; at 21th day, both Cbfal and BMP- 2 were significantly higher than those of control group; at 28th day, the expression of BMP- 2 was still markedly higher compared with control group. Conclusion Lovastatin can promote the osteogenic differentiation of BMSCs, regulate the mRNA expression levels of Cbfal and BMP- 2 at the different phase of osteogenic differentiation, and it may be participate in this progress.