大豆异黄酮与大鼠成骨细胞BMP2PCR产物表达
Effect of soybean isoflavonet on bone morphogenetic protein 2 expression of cultured osteoblasts from rat
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摘要:目的 探讨大豆异黄酮对体外培养大鼠成骨细胞骨形态发生蛋白2(BMP 2)表达的影响。方法 制作新生大鼠颅骨成骨细胞体外培养模型, 用不同浓度的大豆异黄酮刺激分离培养的大鼠成骨细胞, 提取细胞总RNA, RT-PCR扩增BMP 2基因cDNA, 同时扩增β-actin cDNA作为内对照, 扫描RCR产物电泳照片, 计算BMP-2/β-actincDNA的积分吸光度比值, 推算BMP 2基因相对表达水平; 并通过免疫组化方法观察成骨细胞BMP 2的表达, 用HPIAS-2000图象分析仪对各组表达进行定量分析。结果 大豆异黄酮增加体外培养成骨细胞BMP 2的水平, 并且在基因转录水平促进大鼠成骨细胞BMP 2表达, 呈剂量-效应关系(P < 0.01)。结论 大豆异黄酮对成骨细胞形成BMP 2表达有显著促进作用。提示大豆异黄酮促进成骨细胞增殖、分化的作用可能与通过增加BMP 2基因的表达有关。Abstract:Objective To study the effect of soybean isoflavone on bone morphogenetic protein 2(BMP 2)expression of cultured osteoblasts from rat.Methods The osteoblstic cells from cranial bone of newborn rat was cultured.Cultured rat osteoblasts were treated with soybean isoflavone in difierent concentrations.Total RNA was extracted from the cells.BMP 2 were amplified by RT-PCR.β-actin cDNA was also amplified at the same time and used as an internal standard.The SP method was used to detect the effect of soybean isoflavone on BMP 2 expression of cultured osteoblasts.The expression level was monitored with HPIAS-2000 photograph analysis meter.The pictures of PCR production was scanned.The specific values of density of BMP 2/β-actin were calculated, and the expression levels of BMP 2 were determined.Results Soybean isoflavone increased the level of BMP 2 in rat osteoblasts and increased the expressions of BMP 2 genes at the level of transcripaon and revealed dose-effect relation(P < 0.01).Conclusion The soybean isoflavone induces the proliferation and differentiaion of osteoblast probably by increasing the expressions of BMP 2 genes.
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