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目的 :研究骨髓基质细胞 (MSCs)在向成骨细胞(OB)分化的条件培养体系中 ,17β 雌二醇 (E2 )对其核结合因子α1(corebindingfactorα1,Cbfα1)基因表达的影响 ,探讨E2对OB生成的作用及绝经后高转换骨代谢的可能机制 .方法 :取自 3mo龄雌性SD大鼠MSCs在生长培养基中传代后 ,用1,2 5 (OH) 2 D3 和地塞米松 (DEX)诱导MSCs向OB分化 .应用半定量RT PCR技术 ,观察不同浓度E2 对MSCs分化过程中Cbfα1mRNA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MSCs分化过程中Cbfα1mR NA的表达 ,从 (2 3.4± 1.8) %降至 (15 .8± 1.5 ) %和 (5 .8± 0 .8) % (P <0 .0 5 ) ;细胞ALP活性随E2 浓度增加而降低 ,从(70 6± 37)nkat·g-1(protein)降至 (6 3± 5 )nkat·g-1(P <0 .0 5 ) .E2 降低细胞Ⅰ型胶原的合成 ,用VanGieson染色细胞 ,随着E2 浓度的增加 ,胞质染色由鲜红、淡红到黄色 .结论 :E2能明显抑制MSCs分化过程中Cbfα1mRNA的表达 ,减少OB的生成 .
OBJECTIVE: To study the effect of 17β estradiol (E2) on the gene expression of nuclear factor-α1 (Cbfα1) in bone marrow stromal cells (MSCs) differentiated into osteoblast (OB) OB and the possible mechanism of high turnover bone metabolism in postmenopausal women.Methods MSCs from 3-month-old female Sprague-Dawley rats were passaged in growth medium with 1,25 (OH) 2 D3 and dexamethasone (DEX ) Induced the differentiation of MSCs into OB.Using semi-quantitative RT-PCR technique, we observed the effect of different concentrations of E2 on the expression of Cbfα1mRNA during the differentiation of MSCs. The activity of alkaline phosphatase (ALP) The expression of type Ⅰ collagen was detected by staining.Results E2 could significantly inhibit the expression of Cbfα1mRNA in the differentiation of MSCs from (2.34 ± 1.8)% to (15.8 ± 1.5)% and (5.8 ± 0.5)%, respectively 8)% (P <0.05). ALP activity decreased with the increase of E2 concentration from (70 6 ± 37) nkat · g-1 to (6 3 ± 5) nkat · g -1 (P <0.05) .E2 reduced the type I collagen synthesis, with VanGieson staining cells, with the increase of E2 concentration, cytoplasmic staining from bright red, light red to Color Conclusion:. E2 could inhibit the expression during the differentiation of MSCs Cbfα1mRNA reduce generation OB.