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目的:研究骨髓基质细胞向成骨细胞分化过程中,雌二醇对其骨形态发生蛋白受体(BMPR)ⅠA,ⅠBmRNA表达的影响,探讨雌二醇对成骨细胞生成的作用,试阐明绝经后高转换骨代谢始动环节的可能机制。方法:SD大鼠骨髓基质细胞在生长培养基中传代培养后,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,17β-雌二醇(E2)处理培养细胞,观察E2对骨髓基质细胞分化过程中骨形态发生蛋白受体ⅠA、ⅠBmRNA、碱性磷酸酶(ALP)活性和Ⅰ型胶原合成的影响,β-actin作内对照半定量RT-PCR分析骨形态发生蛋白受体ⅠA,ⅠBmRNA的表达,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BMPR-ⅠAmRNA的表达,且呈剂量依赖性,随E2浓度增加(0～1×103nmol/L)BMPR-ⅠAmRNA从(27.0±3.4)%降至(17.0±1.8)%(t=5.2,P<0.01)和(9.3±1.6)%(t=9.2,P<0.01);BMPR-ⅠBmRNA随E2浓度增加而增加,从(2.0±0.8)%增至(4.8±1.5)%(t=3.3,P<0.05)和(17.2±2.2)%(t=13,P<0.01)。Northernblot结果显示在上述E2浓度时BMPR-ⅠAmRNA的表达从4.21±0.36降至1.24±0.10(t=18.2,P<0.01);BMPR-ⅠBmRNA的表达从1.72±0.11增至3.73±0.31(t=12.2,P<0.01)。ALP活性随E2浓度增加而?
OBJECTIVE: To investigate the effect of estradiol on the expression of ⅠA and ⅠB mRNA of bone morphogenetic protein receptor (BMPR) during the differentiation of bone marrow stromal cells into osteoblasts and to explore the effect of estradiol on osteoblastogenesis, After high conversion of bone metabolism may be the first part of the mechanism. METHODS: After SD rat bone marrow stromal cells were subcultured in growth media, bone marrow stromal cells differentiated into osteoblasts with 1,25 (OH) 2D3 and dexamethasone (DEX), 17β-estradiol (E2) The cultured cells were treated with E2 to observe the effect of E2 on bone morphogenetic protein receptor Ⅰ A, Ⅰ B mRNA, alkaline phosphatase (ALP) activity and type Ⅰ collagen synthesis in bone marrow stromal cells. Β-actin was used as an internal control semi-quantitative RT-PCR The expression of bone morphogenetic protein receptor Ⅰ A and Ⅰ B mRNA was analyzed. The activity of alkaline phosphatase was measured by α-phosphoketolacin as substrate. The content of type Ⅰ collagen was detected by Van Gieson staining. Results: E2 significantly inhibited the BMPR-ⅠA mRNA expression in bone marrow stromal cells in a dose-dependent manner. With the increase of E2 concentration (0 ~ 1 × 103nmol / L), BMPR-ⅠA mRNA decreased from (27.0 ± 3.4)% to BMPR-ⅠB mRNA increased with the increase of E2 concentration from (2.0 ± 0.8)% to (17 ± 1.8)% (t = 5.2, P <0.01) and (9.3 ± 1.6)% (4.8 ± 1.5)% (t = 3.3, P <0.05) and (17.2 ± 2.2)% (t = 13, P <0.01). Northern blot results showed that the expression of BMPR-ⅠA mRNA decreased from 4.21 ± 0.36 to 1.24 ± 0.10 (t = 18.2, P <0.01) at the above concentration of E2; the expression of BMPR-ⅠB mRNA increased from 1.72 ± 0.11 to 3.73 ± 0.31 (t = 12.2 , P <0.01). ALP activity increases with E2 concentration?