论文部分内容阅读
目的 研究在MG 6 3细胞和成人成骨细胞培养的不同阶段 ,雌激素对护骨素 (OPG)基因mRNA表达的影响 ,探讨雌激素对成骨细胞的作用机制。方法 在细胞培养的不同时间 ,测定细胞内碱性磷酸酶 (ALP)活性、骨钙素 (OC)含量和进行VanGieson胶原染色 ,以确定细胞培养的增殖、成熟、矿化阶段 ,于成熟阶段用 17β 雌二醇 (E2 )对培养细胞进行处理 ,用半定量RT PCR技术测定雌二醇对成骨细胞OPG基因mRNA表达的影响。结果 ALP活性、骨钙素含量及胶原染色均表明 ,细胞培养汇片后 12天达到成熟。正常成人髂骨分离的成骨细胞 (HOB)及成骨肉瘤细胞株MG 6 3均能表达OPG ,且表达量与培养的时程有关。培养 12天时 ,两种细胞OPG基因mRNA表达均达最高峰 (P <0 .0 1)。 10 -8mol/L的E2 能使MG 6 3细胞OPG基因mRNA的表达增加至基础值的 10倍 (P<0 .0 0 1) ,HOB的OPG基因mRNA表达增加至基础值的 7～ 8倍 (P <0 .0 1)。结论 OPG在成骨细胞表达。雌激素使成骨细胞OPG基因mRNA表达的增加是雌激素缺乏导致骨质疏松的重要机制之一
Objective To study the effect of estrogen on mRNA expression of osteoprotegerin (OPG) in different stages of MG 6 3 cells and adult osteoblasts and to explore the mechanism of action of estrogen on osteoblasts. Methods Intracellular alkaline phosphatase (ALP) activity, osteocalcin (OC) levels and VanGieson collagen staining were determined at various times of cell culture to determine the proliferation, maturation and mineralization stages of the cell culture at the maturation stage 17β-estradiol (E2) was used to treat cultured cells. The effect of estradiol on OPG gene mRNA expression in osteoblasts was determined by semi-quantitative RT-PCR. Results ALP activity, osteocalcin content and collagen staining both showed that the cells reached maturity 12 days after culture. Normal adult iliac osteoblast (HOB) and osteogenic sarcoma cell line MG 6 3 can express OPG, and the expression level and culture schedule. When cultured for 12 days, the mRNA expression of OPG reached the peak in both cells (P <0.01). 10 -8 mol / L of E2 increased the expression of OPG gene mRNA in MG 6 3 cells to 10 times of the basal value (P <0.01), and increased the mRNA expression of OPG gene in HOB to 7 to 8 times of the basal value (P <0. 01). Conclusion OPG is expressed in osteoblasts. Estrogen increases osteoblast OPG gene mRNA expression is one of the important mechanisms of estrogen deficiency leading to osteoporosis