淫羊藿苷对大鼠成骨细胞增殖及碱性磷酸酶、Runx2蛋白表达的影响  

Effect of Icariin on osteoblast proliferation and expression of alkaline phosphatase and Runx2 protein in rats

作　　者：李玲慧[1,2] 丁道芳[1,2] 杜国庆[1,2] 宋奕[1,2] 石印玉[1,2] 詹红生[1,2] 

机构地区：[1]上海中医药大学附属曙光医院石氏伤科医学中心,上海201203 [2]上海市中医药研究院骨伤科研究所,201203

出　　处：《中国矫形外科杂志》2013年第21期2195-2199,共5页Orthopedic Journal of China

基　　金："中医骨伤科学"国家重点学科(编号:100508);国家自然基金项目(编号:81073114;81001527;81102603;81173621);上海领军人才项目(编号:041);上海高校"中医脊柱病损研究"创新团队建设项目(编号:2009-26);上海市科委重点项目(编号:09dZ1973800;08411950300;11DZ1973400;11XD1404600)~~

摘　　要：[目的]探讨淫羊藿苷对体外培养的大鼠成骨细胞增殖及碱性磷酸酶、Runx2蛋白表达的影响。[方法]新生SD大鼠10只,脱臼处死后取出颅盖骨,多次胶原酶消化法分离大鼠成骨细胞。细胞随机分为4组,分别以终浓度为100、50、25μmol/L和0μmol/L的淫羊藿苷干预培养,24、48 h及72 h后采用CCK-8法检测细胞增殖率;加药干预72 h后进行碱性磷酸酶染色,Western Blot法检测PCNA、Runx2蛋白表达量。[结果]加药干预24、48 h后,淫羊藿苷对成骨细胞增殖情况并无显著影响(P>0.05);加药72 h后,100μmol/L组吸光度值显著降低(P<0.05)。各加药组碱性磷酸酶表达量均较对照组有所增高,50μmol/L组及25μmol/L组较明显。Western Blot检测结果显示各浓度淫羊藿苷均可上调Runx2蛋白表达,而各加药组间差异不明显。[结论]淫羊藿苷对体外培养的大鼠成骨细胞增殖并无显著影响,但可明显促进成骨细胞的分化、上调Runx2蛋白的表达;高浓度淫羊藿苷抑制成骨细胞生长。[ Objective] To investigate the effect of Icariin on osteoblast proliferation and expression of alkaline phosphatase and Runx2 protein in rats. [Methods] A total of ten neonatal Sprague - Dawley （SD） rats were killed by dislocation of the neck. Their craniums were removed, and osteoblasts were isolated by type I coUagenase digestion. The cells were randomly divided into four groups and treated with Icariin at concentrations of 100, 50, 25, and 0 μmol/L, respectively. The CCK - 8 method was used to evaluate the osteoblast proliferation at 24, 48, and 72 h after administration of the medication. Alkaline phosphatase staining was performed after 72 h of medication, and western blotting was used to detect PCNA and Runx2 protein expressions. [ Results] The 24- and 48 -h drug interventions had no obvious influence on the osteoblast proliferation （P 〉 0.05） . After 72 h of medication, the odds ratio in the 100 μmol/L group decreased significantly （P 〈 0. 05） . The alkaline phosphatase expression level was higher in the three Icariin groups than in the control group, especially in the 50 and 25 μmol/ L groups. The result of the western blotting analysis showed that Icariin could promote Runx2 protein expression, with no significant changes in the three groups with different concentrations of Icariin. [ Conclusion ] The Icariin administration at different concentrations had no obvious effect on osteoblast proliferation but promoted the osteogenic differentiation of the osteoblasts and upregulated the Runx2 protein expression. However, the high Icariin concentrations inhibited the osteoblast proliferation.

关 键 词：淫羊藿苷 成骨细胞 碱性磷酸酶 RUNX2 

分 类 号：R285.5[医药卫生—中药学]