金雀异黄酮对体外培养成骨细胞抗缺氧的机制研究

Study on Mechanism of the Anti Hypoxia Effect of Genistein on Osteoblasts in vitro

目的:观察金雀异黄酮对缺氧成骨细胞的抗缺氧作用。方法:以大鼠成骨细胞为材料,用三气培养箱建立缺氧模型。设常氧对照组、缺氧对照组和含不同浓度金雀异黄酮的缺氧加药组。比较各组缺氧36 h后细胞活力、ROS含量、细胞凋亡、细胞周期、PCNA表达i、NOS活性和钙化结节面积等,荧光定量PCR检测HIF-1α、BCL-2及Caspase-3 mRNA的表达。结果:金雀异黄酮显著提高缺氧成骨细胞存活率、G1期细胞百分比、钙化结节面积、HIF-1α和BCL-2 mRNA的表达水平,降低凋亡、ROS含量、PCNA表达量i、NOS酶活性和Caspase-3 mRNA表达。结论:金雀异黄酮能保护缺氧成骨细胞并促进其分化。

Objective：To study the protective effect of genistein on osteoblasts treated with hypoxia. Methods：Rat osteoblasts were isolated from calvarias of newborn Sprague-Dawly rat by enzyme digestion and hypoxic environment was made by triple-gases incubator. Rat osteoblasts treated with hypoxia for 36 haurs. After 36 hours, cell viability, content of reactive oxygen species （ ROS）, analysis of cellular cycle and apoptosis, expression of proliferating cell nuclear antigen（ PCNA）, activity of iNOS and area of calcified nodules were detected. Total RNA was isolated and the gene expression of hypoxia inducible factor-1 a（ HIF-1α）, BCL-2 and Caspase-3 was investi- gated by Real Time RT-PCR. Results：Genistein could significantly improve cell viability, percentage of G1 phases, area of calcified nod- ules and decrease apoptosis rate, ROS content, expression of PCNA, activity of iNOS. Besides, mRNA levels of HIF-1 α and BCL-2 were enhanced and that of Caspase-3 was inhibited. Conclusion：Genistein can protect osteoblasts from hypoxia and enhance osteogenic dif- ferentiation significantly.