芒柄花素对体外培养成骨细胞缺氧损伤的保护作用  

Protective effect of formononetin on hypoxic injuries of osteoblasts in vitro

作　　者：韩桂秋[1] 王鸣刚[1] 陈克明[2] 葛宝丰[2] 马慧萍[3] 周建[2] 明磊国[2] 朱瑞清[2] 

机构地区：[1]兰州理工大学生命科学与工程学院 [2]兰州军区兰州总医院骨科研究所 [3]兰州军区兰州总医院药材科,甘肃兰州730050

出　　处：《中国药理学通报》2011年第5期671-677,共7页Chinese Pharmacological Bulletin

基　　金：甘肃省科技重大专项计划资助项目(No092nkDA025)

摘　　要：目的研究芒柄花素在缺氧培养条件下对成骨细胞增殖、细胞周期、凋亡和成骨分化等的影响。方法采用酶消化法分离培养SD大鼠颅骨成骨细胞,并用三气培养箱建立缺氧模型。将细胞分为常氧对照组、缺氧对照组和缺氧加药组,其中缺氧加药组进一步分为10-6、10-5和10-4 mol·L-1组,分别加入10-6、10-5和10-4 mol·L-1芒柄花素。于缺氧处理36 h后分析各组的细胞存活率、LDH漏出率、细胞凋亡、细胞周期等,并用RT-PCR法检测缺氧诱导因子-1α、Bcl-2及Caspase-3的基因表达情况。缺氧处理48 h后检测ALP活性、钙化结节面积等成骨分化指标。结果与缺氧对照组比较,芒柄花素可提高细胞存活率,降低LDH漏出率,减少细胞凋亡并提高G1期细胞百分比,提高HIF-1α和Bcl-2mRNA的表达水平,抑制Caspase-3 mRNA的表达水平,对ALP活性和钙化结节面积等也有提高作用,且均呈现出剂量依赖性特点。结论芒柄花素对成骨细胞缺氧损伤具有明显保护作用。Aim To investigate the effects of formononetin on survival rate,cellular cycle,apoptosis and differentiation of osteoblasts induced by hypoxia.Methods Rat osteoblasts were isolated from calvarias of newborn Sprague-Dawly by enzyme digestion,and hypoxic environment was made by triple-gases incubator.Rat osteoblasts was induced by hypoxia.After 36 hours,cell viability,LDH leakage rate,analysis of cellular cycle and apoptosis rate were compared among 10-6 mol·L^-1 group,10-5 mol·L^-1 group,10-4 mol·L^-1 group,hypoxia control and normal control.Total RNA was isolated and the gene expression of HIF-1α,Bcl-2 and Caspase-3 was investigated by RT-PCR.48 hours after hypoxia,osteogenic differentiation markers including ALP activity and nodules were detected.Results Formononetin could significantly improve survival rate,ALP activity,area of calcified nodules and decrease LDH leakage rate,apoptosis,percentage of S＋G2 phases.Besides,the mRNA level of HIF-1α and Bcl-2 was enhanced,and the mRNA level of Caspase-3 was inhibited.Conclusion Formononetin can protect osteoblasts from hypoxia and enhance the osteogenic differentiation of osteoblasts induced by hypoxia significantly.

关 键 词：缺氧诱导因子-1Α 芒柄花素 成骨细胞 细胞周期 凋亡 Bcl-2 CASPASE-3 

分 类 号：R-332[医药卫生] R282.71