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目的观察2β(3羟丙氧)骨化三醇(ED71)对体外培养人成骨样细胞增殖与分化的影响,进一步探讨ED71治疗骨质疏松的机制。方法采用塞唑蓝(MTT)分析法检测ED71对体外培养OS732人成骨样细胞增殖作用与效价。采用培养细胞匀浆测单位蛋白碱性磷酸酶活性,培养基中骨钙素含量分析及原位杂交方法检测ED71对OS732人成骨样细胞分化的影响。结果培养第4天,10-7mol/LED71显著抑制成骨样细胞增殖(P<0.05),而10-8mol/L、10-9mol/LED71对成骨样细胞的作用不明显。培养4d后培养基中骨钙素水平在10-7mol/LED71组变化不大,在10-8mol/L、10-9mol/LED71组骨钙素水平显著增高(P<0.05或P<0.01)。原位杂交转化生长因子β(TGFβ1)表达明显增加,而碱性磷酸酶比活性无明显变化。结论ED71抑制成骨样细胞增殖高剂量作用显著,促进成骨样细胞分化较低剂量显著。
Objective To observe the effect of 2β (3-hydroxypropyloxy) calcitriol (ED71) on the proliferation and differentiation of human osteoblast-like cells in vitro and to explore the mechanism of ED71 in the treatment of osteoporosis. Methods The proliferative effect and titer of ED71 on OS732 human osteoblast-like cells were detected by MTT assay. The effect of ED71 on the differentiation of OS732 human osteoblast-like cells was examined by measuring the activity of alkaline phosphatase (ALP), the content of osteocalcin in culture medium and the method of in situ hybridization. Results On day 4, 10-7mol / LED71 significantly inhibited the proliferation of osteoblast-like cells (P <0.05). However, the effect of 10-8mol / L and 10-9mol / LED71 on osteoblast-like cells was not obvious. After cultured for 4 days, the level of osteocalcin in the medium of 10-7mol / LED71 did not change much. The level of osteocalcin in 10-8mol / L, 10-9mol / LED71 group was significantly increased (P <0.05 or P <0.01). In situ hybridization TGFβ1 expression increased significantly, while the alkaline phosphatase specific activity did not change significantly. Conclusion ED71 can inhibit the proliferation of osteoblast-like cells at a high dose and promote the differentiation of osteoblast-like cells at a lower dose.