【摘要】 目的筛选双氧水可以引起哪些骨生成相关基因的改变。方法用胰岛素-转铁蛋白和亚硒酸钠的混合液诱导鼠软骨前体细胞分化为肥大软骨细胞。采用MTT法确定双氧水作用的最佳浓度和时间。采用PCR阵列检测细胞中84个骨生成相关基因的表达;并用定量RT-PCR验证PCR阵列结果。结果结果显示:9个基因表达上调、12个基因表达下调,这些基因编码多种功能蛋白,其中包括胶原蛋白、转录因子、骨骼发育和骨矿物质代谢相关蛋白以及细胞粘附分子等。定量RT-PCR验证了5个表达下调基因(Smad2,Smad4,转化生长因子βr1、βr3,以及基质金属蛋白酶10)。结论双氧水改变了一些具有不同生物学功能的基因在肥大软骨细胞中的表达。结合氧化损伤与Kashin-Beck病相关的文献,我们推测这些基因有可能参与了Kashin-Beck病软骨的深层坏死。更多还原

【Abstract】 Objective To identify the osteogenesis genes whose expression is altered in hypertrophic chondrocytes treated with H2 O2.Methods Murine chondrogenitor cells(ATDC5) were differentiated into hypertrophic chondrocytes by InsulinTransferrin-Selenium(ITS) treatment, and then treated with H2 O2. Suitable conditions(concentration, time) were determined by using the MTT assay. After total RNA isolation and cD NA synthesis, the levels of 84 genes were determined using the PCR array, whereas quantitative RT-PCR was carried out to validate the PCR array data. Results We identified 9 up-regulated genes and 12 down-regulated genes, encoding proteins with various functions, such as collagen proteins, transcription factors, proteins involved in skeletal development and bone mineral metabolism, as well as cell adhesion molecules. Quantitative RT-PCR confirmed the altered expression of 5 down-regulated genes(Smad2, Smad4, transforming growth factor β receptor 1, transforming growth factor β receptor 3, and matrix metalloproteinase 10). Conclusions H2 O2 significantly changed the expression of several genes involved in a variety of biological functions. Because of the link between oxidative damage and Kashin-Beck disease, these genes may also be involved in the deep-zone necrosis of the cartilage observed in Kashin-Beck disease.更多还原