摘要
目的研究骨髓基质细胞在向成骨细胞分化的介质中，17p雌二醇（E2）对其骨形态发生蛋白受体ⅠB（BMPR-ⅠB）及过氧化物酶增殖活化受体γ2（PPAR-γ2）mRNA表达的影响，探讨雌二醇对骨髓基质细胞分化的作用。方法1,25（OH）2D3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Northern blot技术。观察不同浓度E2对骨髓基质细胞分化过程中BMPR-ⅠB及PPAR-γ2 mRNA表达的影响；以α-磷酸簪酚为底物。测定细胞碱性磷酸酶的活性，Van Gieson染色法显示Ⅰ型胶原的含量。结果E2能明显增强骨髓基质细胞分化过程中BMPR-ⅠBmRNA的表达，E2浓度为（0～10^-6）moL／L时。BMPR-ⅠBmRNA的表达随E2浓度增加而增加，从（2．0±0．4）％增至（4．8±1．0）％（P〈0．05）和（17．3±2．2）％（P〈0．01）；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（1．80±0．3）％增至（9.5±2.1）％（P〈0．01）和（19．2±3．0）％（P〈0．01）；Northern blot结果显示随着E2浓度的增加，BMPR-ⅠBmRNA的表达从1．72±0．11增至3．73±0．31（P〈0．01）；PPAR-γ2mRNA的表达量从1．47±0．12增至2．81±0．22（P〈0．01）和4．02±0．36（P〈0．01）。细胞ALP活性明显受E2的抑制，在上述E2浓度时ALP的活性从（42．6±2．5）降至（3．6±0．7）U／g蛋白（P〈0．01）。Ⅰ型胶原含量均随E2浓度增加而降低。结论E2抑制体外培养的骨髓基质细胞向成骨细胞分化而促进其向脂肪细胞分化。
Objective To investigate the effects of 17β-estradiol（E2） on the gene expression of bone morphogenetic protein receptor ⅠB （BMPR- ⅠB） and peroxisome proliferator-activated receptor gamma 2（PPAR-7） in rat bone marrow stromal cells exposured to the differentiation medium and to elucidate the role of E2 on differentiation of these ceils. 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）2D310^-9 mol/L and different concentrations （0 -10^- 6mol/L） of E2. Total cellular RNA was isolated using the total RNA kit.The effects of E2 on BMPR- ⅠB and PPAR-γ2 gene expression were assayed by semiquantitative RT-PCR and demonstrated by Northern blot, respectively. Results E2 strongly raised the expression of BMPR- ⅠB mRNA in bone marrow stromal cell .The increase was dose dependent. When examined under various concentrations of E2 （0 - 10^-6mol/L）, the expression of BMPR- ⅠB mRNA were increased from 2.0% ± 0.4% to 4.8% ± 1.0% （ P 〈 0.05） ,8.0% ± 1.4% （ P 〈 0.01 ） and 17.3% ± 2.2% （P 〈 0.01）. E2 strongly stimulated the expression of PPAR-72 mRNA in bone marrow stromal cell from （1.80±0.3）% to （9.5±2.1）% （P〈0.01） and （19.3±3.0）% （P〈0.01）.Northern blot showed that the expression of BMPR- ]. B and PPAR-72 mRNA in bone marrow stromal cell was increased by E2 from 1.72 ± 0.11 to 3.73 ± 0.31 （ P 〈 0.01 ） and from 1.47 ± 0.12 to 4.02 ± 0.36（ P 〈 0.01 ） ,respectively .The activities of ALP suppressed from （42.6 ± 2.5 to 3.6 ± 0.7） U/g protein （ P 〈 0.01 ） by E2. The amount of type Ⅰ collagen were decreased significantly at higher concentrations of E2. Conclusions E2 promoted bone marrow stromal ceils differentiating into adipocytes and inhibited osteoblastogenesis in vitro.
出处
《西南军医》
2006年第1期4-7,共4页
Journal of Military Surgeon in Southwest China
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
骨形态发生蛋白受体ⅠB
17β-estradiol bone marrow stromal cell peroxisome proliferator-activated receptor （PPAR-γ2） differentiation bne morphogenetie protein receptor ⅠB（ BMPR-Ⅰ B）
