阿伦膦酸钠对去势大鼠骨髓间充质干细胞成脂分化的影响

摘要

Objective To explore the effects of alendronate on adipogenic differentiation of bone marrow stremal cells (BMSCs) and the role of mitogen-activated protein kinases (MAPK) signal pathway in this process. Methods BMSCs were derived from 9-month-old ovariectomized SD rats and exposed to 0.01, 0.1, 1, 10 μmol/L of alendronate for 2 weeks. The number of BMSCs was counted under light microscope after oil red O staining. The expression of peroxisome proliferators activated receptor-γ, 2 (PPAR-γ2) was measured by RT-PCR. The effect of alendronate on MAPK signal pathway was detected by Western blot. Results After two weeks of induction of BMSCs by alendronate, BMSCs with positive oil red O staining significantly decreased as the increase of the concentration of alendronate (P <0.01), so did the expression of PPAR-γ2. The expression level of PPAR 2 increased when exposing BMSCs to ERK1/2 or JNK specific inhibitors, PD98059 and SP600125 for two weeks. However, the expression level of PPAR 2 decreased when exposing BMSCs to SB203580 (an inhibitor of p38) for two weeks. Condusion Alendronate can inhibit adipogenic differentiation of BMSCs derived from ovariectomized rats in a doso-dependent manner by activating ERK1/2 and JNK rather than p38.%目的 探讨阿伦膦酸钠(Alendronate,Aln)对骨髓间充质干细胞(BMSCs)成脂分化的影响,并阐明丝裂原活化蛋白激酶(MAPK)信号通路在该过程中的作用. 方法 BMSCs取自9个月龄去势SD大鼠,分别暴露于0.01,0.1,1,10 μmol/L Aln.成脂诱导2周后进行油红O染色和镜下计数分析,RT-PCR检测过氧化物酶体增殖体激活受体γ2(PPARγ2)表达;使用MAPK通路特异性抑制剂并诱导2周后再观察PPARγ2表达情况,Western blot观察Aln对MAPK通路表达的影响. 结果 BMSCs成脂诱导2周后,油红O染色阳性细胞数随着药物浓度的增高而显著减少(P<0.01).PPARγ2表达随着药物浓度的增高而显著降低.分别使用ERK1/2、JNK抑制剂PD98059和SP600125并诱导2周后,PPARγ2表达上调;使用p38抑制剂SB203580并诱导2周后,PPARγ2表达下调.Western blot结果显示,Aln在5,15,30 min时上调P-ERK1/2和P-JNK的表达,分别使用抑制剂后,表达下调. 结论 Aln通过激活ERK1/2和JNK信号通路,而不是p38,发挥抑制去势大鼠来源的BMSCs成脂分化作用,其效应具有浓度依赖性.