摘要
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 .
AIM: To investigate the effects of 17β estradiol (E 2) on the gene expression of core binding factor alpha 1 (Cbf α1) during the differentiation of rat bone marrow stromal cells exposed to the differentiation medium and to elucidate the effects of E 2 on the osteoblastogenesis. METHODS : Adherent bone marrow stromal cells obtained from 3 month old female SD rat and cultured in growth medium were treated at 37℃, 50 mL·L -1 CO 2 for 3 days in the presence of DEX 1×10 -7 mol·L -1 and 1,25 (OH) 2D 3 1×10 -9 mol·L -1 and with different concentrations of E 2. Total cellular RNA was isolated using total RNA kit. The effects of E 2 on the Cbf α1 gene expression were quantified by semiquantitative RT PCR. RESULTS: E 2 strongly inhibited the expression of Cbf α1 mRNA during the differentiation of bone marrow stromal cells from (23.4±1.8)% to (15.8± 1.5)% and (5.8±0.8)%( P <0.05). The suppression of the Cbf α1 mRNA was dose dependent. The activity of ALP decreased significantly with the increase of E 2 concentration, from (706±37) nkat·g -1 (protein) to(63±5) nkat· g -1 (protein) ( P <0.05). The amount of type Ⅰcollagen was suppressed by E 2. CONCLUSION: E 2 markedly inhibits Cbf α1 gene expression during the differentiation of bone marrow stromal cells and reduces the osteoblastogenesis.
出处
《第四军医大学学报》
北大核心
2003年第13期1227-1230,共4页
Journal of the Fourth Military Medical University
