红车轴草提取物对破骨细胞骨吸收及分化的作用机制  

Effect of Trifolium pratense L. extracts on the bone resorption and osteogenic differentiation of osteoclasts and the underlying mechanisms

作　　者：李扬[1] 王德平[2] 艾东[3] 魏鑫[3] 关利新[3] 关悦[2] 

机构地区：[1]牡丹江医学院附属红旗医院骨科,黑龙江省牡丹江市157011 [2]牡丹江医学院附属红旗医院内分泌科,黑龙江省牡丹江市157011 [3]牡丹江医学院,黑龙江省牡丹江市157011

出　　处：《中国组织工程研究》2017年第20期3129-3134,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:最近研究发现红车轴草异黄酮能够通过提高雌激素作用,有效抑制骨吸收,降低骨转化率,增强成骨细胞活性及骨密度,对骨质疏松具有一定的防治作用。目的:观察红车轴草提取物对破骨细胞骨吸收及分化的作用影响。方法:提取大鼠骨髓腔细胞,用淋巴细胞分离液分离细胞,培养5 h后,取未贴壁细胞加入30μg/L巨噬细胞集落刺激因子和75μg/L核因子κB受体活化因子配体诱导细胞(对照组),将不同质量浓度(0.3,0.6和1.2 g/L)的红车轴草提取物加入诱导的破骨细胞中,观察不同质量浓度的红车轴草提取物对破骨细胞分化与骨吸收作用;采用Western bloting法测定破骨细胞分化相关蛋白c-fos和NFATcl的含量。结果与结论:(1)与对照组比较,不同质量浓度的红车轴草提取物能够不同程度的抑制破骨细胞分化和骨吸收;(2)抗酒石酸酸性磷酸酶染色发现红车轴草提取物能够明显降低破骨细胞数目;(3)Western blotting结果表明红车轴草提取物能够抑制破骨细胞分化相关蛋白c-fos和NFATC1表达;(4)结果提示,红车轴草提取物能够抑制破骨细胞分化及骨吸收。BACKGROUND： Isoflavone isolated from Trifolium pratense L. has been found to be able to effectively inhibit bone resorption, reduce bone turnover rate, improve osteocyte activity and bone mineral density by enhancing the effect of estrogen, which is helpful for the prevention and treatment of osteoporosis. OBJECTIVE： To investigate the effect of Trifolium pratense L. extracts on the bone resorption and differentiation of osteoclasts. METHODS： Rat bone marrow cells were extracted, isolated by lymphocyte separation and cultured for 5 hours; then, the non-adherent cells were selected followed by induced by 30 μg/L macrophage colony stimulating factor and 75 μg/L RANKL（control groups）, or different concentrations of Trifolium pratense L. extracts（0.3, 0.6 and 1.2 g/L） to observe their effect on the osteoclast differentiation and bone resorption. The levels of osteoclast differentiation-associated proteins c-fos and NFATcl were determined by western blot assay. RESULTS AND CONCLUSION： Compared with the control group, different concentrations of Trifolium pratense L. extracts could suppress osteoclast differentiation and bone resorption to different degrees. Tartrate-resistant acid phosphatase staining showed that Trifolium pratense L. extracts could significantly reduce the number of osteoclasts. Western blot assay results suggest that Trifolium pratense L. extracts significantly inhibited the expression levels of c-fos and NFATcl. These results reveal that Trifolium pratense L. extracts can inhibit osteoclast differentiation and bone resorption.

关 键 词：破骨细胞 细胞分化 百脉根属 组织工程 组织构建 骨组织工程 骨质疏松 破骨细胞分化 骨吸收 抗酒石酸酸性磷酸酶 骨陷窝 NFATC1 c-fos 红车轴草 

分 类 号：R318[医药卫生—生物医学工程]