摘要
目的 研究骨髓基质细胞在向成骨细胞分化的介质中 ,17β 雌二醇 (E2 )对其核结合因子α1(Cbfα1)及过氧化物酶增殖活化受体γ2 (PPAR γ2 )mRNA表达的影响 ,探讨雌二醇对成骨细胞生成的作用。方法 1,2 5 (OH ) 2 D3 和地塞米松 (DEX)诱导大鼠骨髓基质细胞向成骨细胞分化 ,应用半定量RT PCR及Northernblot技术 ,观察不同浓度E2对骨髓基质细胞分化过程中Cbfα1及PPAR γ2mRNA表达的影响 ,以α 磷酸奈酚为底物 ,测定细胞碱性磷酸酶 (ALP)的活性 ,VanGieson染色法显示Ⅰ型胶原的含量。结果 E2能明显抑制骨髓基质细胞分化过程中Cbfα1mRNA的表达 ,E2浓度为 (0 10 -6)mol/L时 ,Cbfα1mRNA的表达量从 (2 5 0± 3 3 ) %降至 (14 5± 1 3 ) % (P <0 0 1)和 (6 5± 1 9) % (P <0 0 1) ;E2能明显促进骨髓基质细胞在分化介质中PPAR γ2mRNA的表达 ,在上述E2浓度时 ,PPAR γ2mRNA的表达从 (1 75± 0 5 ) %增至 (9 5± 2 1) % (P <0 0 1)和 (19 3± 3 0 ) % (P <0 0 1) ;Northernblot结果显示随着E2浓度的增加 ,CbfαlmRNA的表达量从 4 42± 0 3 9降至 1 88± 0 16(P <0 0 1)和 1 2 0± 0 11(P <0 0 1) ;PPAR γ2mRNA的表达量从 1 47± 0 12增至 2 81±0 2 2 (P <0 0 1)和 4 0 2± 0 3 6(P
Objective To investigate the effects of 17 β estradiol(E2)on the gene expression of core binding factor alpha 1(Cbfα1)and peroxisome proliferator activated receptor gamma 2(PPAR γ2)in rat bone marrow stromal cells exposured to the differentiation medium and elucidate the role of E2 in osteoblastogenesis Methods Adherent bone marrow stromal cells from 3 month old female SD rat were cultured in DMEM supplemented with 10% FBS and dexamethasome(DEX) 10 -7 mol/L and 1,25(OH) 2D 3 10 -9 mol/L and different concentrations(0 10 -6 mol/L)of E2 Total cellular RNA was isolated using the total RNA kit The effects of E2 on the Cbfα1 and PPAR γ2 gene expression were assayed by semiquantiative RT PCR and demonstrated by Northern blot,respectively Results E2 strongly inhibited the expression of Cbfα1 mRNA in bone marrow stromal cell The suppression was dose dependent When examined under various concentrations of E2(0 10 -6 mol/L),the expression of Cbfα1 mRNA decreased from(25 0±3 3)% to(19 8±2 2)%,(14 5±1 3)%( P <0 01) and(6 5±1 9)%( P <0 01) E2 strongly stimulated the expression of PPAR γ2 mRNA in bone marrow stromal cell which increased from(1 75±0 5)% to(9 5±2 1)%( P <0 01) and(19 3±3 0)%( P <0 01) Northern blot showed that the expression of Cbfα 1 in bone marrow stromal cell was decreased by E2 from(4 22±0 39) to(1 20±0 11)( P <0 01) but PPAR γ2 mRNA increased from(1 47±0 12) to(4 02±0 36)( P <0 01) The activities of ALP were suppressed and decreased from(42 6±2 5 to 3 6±0 7)U/g protein( P <0 01) by E2 The amount of type I collagen decreased significantly at higher concentrations of E2 Conclusion E2 promotes bone marrow stromal cells differentiating into adipocytes and inhibits osteoblastogenesis in vitro
出处
《四川医学》
CAS
2003年第11期1105-1108,共4页
Sichuan Medical Journal
关键词
雌二醇
骨髓基质细胞
过氧化物酶增殖活化受体γ2
分化
核结合因子α1
17 β-estradiol
bone marrow stromal cell
peroxisome proliferator-activated receptor(PPAR γ2 )
differentiation
core binding factor alpha 1(Cbfα1)
