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目的研究骨髓基质细胞在向成骨细胞分化的条件培养体系中,17β-雌二醇(E2)对其核结合因子α1(Cbfα1)基因表达的影响,探讨雌二醇对成骨细胞生成的作用。方法取自1只3月龄雌性SD大鼠骨髓基质细胞在生长培养基中传代后,用1,25(OH)2D3和地塞米松(DEX)诱导骨髓基质细胞向成骨细胞分化。应用半定量RT-PCR技术,观察不同浓度E2对骨髓基质细胞分化过程中核结合因子α1mRNA表达的影响,以α-磷酸奈酚为底物,测定细胞碱性磷酸酶的活性,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,且呈剂量依赖性。E2浓度为0,1×10-10,1×10-8和1×10-6mmol/L时,Cbfα1mRNA的表达量分别为23.4%±1.8%,21.8%±2.0%,15.8%±1.5%(t=6.3,P<0.01)和5.8%±0.8%(t=17.8,P<0.01)。Northernblot结果显示Cbfα1mRNA的表达量分别为4.22±0.11,2.51±0.12(t=21,P<0.01),1.88±0.10(t=31,P<0.01)和1.17±0.09(t=41,P<0.01)。ALP活性随E2浓度增加而降低,在上述E2浓度时,ALP活性分别为(42.6±2.5)U/g,(17.9±2.0)U/g(t=15,P<0.01),(10.8±1.5)U/g(t=21,P<0.01)和(3.6±0.7)U/g(t=29,P<0.01)。细胞Ⅰ型胶原的含量随E2浓度增加而降低。结论E2能明显抑制骨髓基质细胞分化过程中Cbfα1mRNA的表达,减少
Objective To investigate the effect of 17β-estradiol (E2) on the expression of Cbfα1 in bone marrow stromal cells under osteoblast differentiation and to explore the effect of estradiol on osteoblastogenesis . METHODS: Bone marrow stromal cells (MSCs) from 3-month-old female Sprague-Dawley rats were passaged in growth medium and then differentiated into osteoblasts by 1,25 (OH) 2D3 and dexamethasone (DEX). Semi-quantitative RT-PCR was used to observe the effects of different concentrations of E2 on the expression of nuclear factor-α1mRNA in bone marrow stromal cells. The activity of alkaline phosphatase was determined by α-plastid phenolphthalein. Van Gieson staining showed that Ⅰ Collagen content. Results E2 could significantly inhibit the expression of Cbfα1 mRNA in bone marrow stromal cells in a dose-dependent manner. The expression levels of Cbfα1 mRNA were 23.4% ± 1.8%, 21.8% ± 2.0% and 15.8% ± 1.5% at E2 concentrations of 0,1 × 10-10, 1 × 10-8 and 1 × 10-6mmol / L, respectively t = 6.3, P <0.01) and 5.8% ± 0.8% (t = 17.8, P <0.01). Northernblot results showed that the expression of Cbfα1 mRNA was 4.22 ± 0.11,2.51 ± 0.12 (t = 21, P <0.01), 1.88 ± 0.10 (t = 31, P <0.01) and 1.17 ± 0.09 ). The activity of ALP decreased with the increase of E2 concentration. The activity of ALP in the above concentration of E2 was (42.6 ± 2.5) U / g, (17.9 ± 2.0) U / g ) U / g (t = 21, P <0.01) and (3.6 ± 0.7) U / g, respectively. The content of type Ⅰ collagen decreased with the increase of E2 concentration. Conclusion E2 can significantly inhibit the expression of Cbfα1 mRNA in bone marrow stromal cells during differentiation, decrease