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目的 研究雌二醇 (17β estradiol,E2 )和抗坏血酸 (VitC)对成骨肉瘤样细胞株MG 6 3的协同作用 ,从而探讨E2 治疗骨质疏松的作用机制。 方法 利用 10 8mol/L的E2 与 10 0 μg/ml的VitC单独或共同处理成骨样细胞株MG 6 3,3 H掺入法 (3 H TdR)测定细胞的增殖 ,α 磷酸奈酚法测细胞内碱性磷酸酶 (ALP)活性的变化 ,半定量逆转录聚合酶链式反应 (RT PCR)测I型胶原 (collagenI)、基质金属蛋白酶 (matrixmetalloproteinase ,MMP) 1、MMP 2及护骨素 (osteoprotegerin ,OPG)基因表达的变化 ,明胶酶谱分析测MMP 2的活性 ,观察E2 及VitC对上述指标的影响。 结果 E2 与VitC合用较单用能使细胞内ALP活性增至对照组的 2倍 (1 0 4± 0 0 5对 1 99± 0 6 2 ,P <0 0 0 5 ) ,3 H TdR掺入量增至对照组的6倍 (4 386± 430对 2 6 5 37± 146 0 ,P <0 0 0 1) ,MMP 1表达降低为原来的 1/ 6 (P <0 0 5 ) ,OPG基因表达增至对照组的 9倍 (P <0 0 1)。明胶酶谱分析显示VitC使MMP 2分泌增加 ,而E2 使其分泌减少 ,二者合用对其无明显影响。 结论 VitC能使E2 作用加强 ,二者协同在绝经后妇女的骨质疏松治疗中发挥重要作用。
Objective To investigate the synergistic effect of 17β estradiol (E2) and ascorbic acid (VitC) on osteosarcoma cell line MG 6 3 to explore the mechanism of E2 treatment of osteoporosis. Methods Osteoblast-like cell line MG 6 3,3 H incorporation (3 H TdR) was used to measure the proliferation of cells by using 10 8 mol / L E2 and 100 μg / ml VitC respectively. Intracellular alkaline phosphatase (ALP) activity was detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT PCR). Collagen I, matrix metalloproteinase 1, MMP 2 and osteoprotegerin (osteoprotegerin, OPG) gene expression changes, gelatin zymography assay MMP 2 activity observed E2 and VitC on the above indicators. Results Compared with single use of VitC, E2 increased the intracellular ALP activity to twice as much as that of the control group (104 ± 0 05 vs. 1 99 ± 0 6 2, P 0 05), 3 H TdR incorporation (4 386 ± 430 vs 2665 37 ± 146 0, P 0 01), and the expression of MMP 1 was reduced to 1/6 (P 0 05). The expression of OPG gene The expression was increased 9-fold in the control group (P <0.01). Gelatin zymography showed that VitC increased MMP 2 secretion, while E2 decreased its secretion, the combination of the two had no significant effect. Conclusion VitC can enhance the role of E2, both of which play an important role in the treatment of osteoporosis in postmenopausal women.