骨髓基质细胞成脂分化及辛伐他汀对其影响的实验研究

Study on the potential and the effect of simvastatin on adipocytic differentiation of bone marrow stromal cells

目的 观察骨髓基质细胞的成脂分化潜能 ,研究辛伐他汀对骨髓基质细胞成脂分化的影响 ,探讨辛伐他汀刺激成骨的作用机制。 方法 体外培养成年小鼠骨髓基质细胞 ,脂肪细胞分化诱导剂 (HI)作用 6d ,检测细胞碱性磷酸酶 (ALP)比活性的变化。HI与不同浓度的辛伐他汀共同作用 72h ,RT PCR检测脂蛋白脂酶 (LPL)mRNA表达水平的变化 ;HI与不同浓度的辛伐他汀或 10 0μg/L重组人骨形态发生蛋白 2 (rhBMP 2 )共同作用 12d后 ,油红O染色、荧光活化的细胞分选(FACS)检测脂肪细胞分化比例。 结果 HI作用 6d后 ,细胞ALP比活性降低 ,约 4 0 3%。分别为( 383 6 1± 134 30 )U·g 1·L 1和 ( 891 5 1± 2 82 5 2 )U·g 1·L 1,P <0 0 1。在含HI的培养液中 ,辛伐他汀作用 72h后 ,LPLmRNA表达水平降低 ;辛伐他汀作用 12d后 ,脂肪细胞的分化比例显著减低 (P<0 0 1)。 结论 骨髓基质细胞可以分化为脂肪细胞 ,并伴随成骨活性减低 ;辛伐他汀抑制骨髓基质细胞的成脂分化 ,辛伐他汀治疗骨质疏松的作用机制可能与此有关。

Objective To observe the adipocytic differentiation potential of bone marrow stromal cells (BMS), and the effect of simvastatin on adipocytic differentiation of bone marrow stromal cells in vitro, and to elucidate the mechanisms of anabolic effect of simvastatin on bone formation. Methods BMS from femur and tibia of adult female BALB C mice were cultured in vitro. Changes of alkaline phosphatase (ALP) activity were determined after treatment with adipogenetic agonist (hydrocortisone 0 5 μmol/L and indomethacin 60 μmol/L, HI) for 6 days. Thenexpression of lipoprotein lipase (LPL) mRNA was detected by RT PCR after treatment with HI and different concentration of simvastatin for 72 h. Adipogenetic differentiation were also observed with Oil Red O staining and fluorescence activated cell sorting (FACS) after treatment with HI and different concentration of simvastatin or 100 μg/L rhBMP 2 for 12 days. Results After BMS were treated with HI for 6 days, ALP activity was significantly decreased ( P <0 01); after BMS were treated with simvastatin and HI for 3 days, the level of LPL mRNA decreased; Oil Red O staining and FACS showed that simvastatin significantly inhibited adipocytic differentiation of BMS( P <0 01). Conclusions BMS can differentiate into adipocyte, accompanied by osteogenesis decreasing; simvastatin inhibits adipocytic differentiation of bone marrow stromal cells in vitro. These may be parts of the mechanism of anabolic effect of simvastatin on bone formation.