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钟海波,郭祥,黄琳惠.葛根素通过ERK1/ 2 和p38 MAPK 信号通路刺激成骨分化和骨形成的机制[J].中国比较医学杂志,2019,29(2):78~83.
葛根素通过ERK1/ 2 和p38 MAPK 信号通路刺激成骨分化和骨形成的机制
Puerarin stimulates osteogenesis and bone formation through ERK1/ 2 and p38 MAPK signaling pathways
投稿时间:2018-06-20  
DOI:10.3969/j. issn. 1671 -7856. 2019. 02. 013
中文关键词:  葛根素  信号通路  骨分化  骨形成
英文关键词:puerarin  signal transduction pathway  bone differentiation  bone formation
基金项目:
作者单位E-mail
钟海波 中南大学湘雅医学院附属海口医院骨科,海口 570208 13907556602@ 139.com 
郭祥 中南大学湘雅医学院附属海口医院骨科,海口 570208  
黄琳惠 海南省人民医院呼吸与危重症医学科,海口 570311  
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中文摘要:
      目的 探究葛根素通过ERK1/2 和p38 MAPK 信号通路刺激成骨分化和骨形成的机制?方法 分离培养成人成骨细胞(MC3T3-E1),运用MTT 法对比加入不同浓度葛根素后细胞增殖能力和对生长曲线的影响?通过测定碱性磷酸酶活性检测葛根素对成骨细胞分化的影响?通过测定钙沉积量分析葛根素对骨形成的影响?通过ERKl/2 阻断剂PD8089,p38 抑制剂SB203580 的加入分析ERK1/2 和p38 MAPK 信号通路刺激成骨分化和骨形成的机制?利用蛋白质免疫印迹(WB)检测细胞骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)的表达?结果 不同浓度葛根素干预成骨细胞,可以不同程度上促进其增殖,1 μmol/ L 葛根素组效果最明显?第1 天和第3 天较空白组增殖趋势不明显,第5 天和第7 天较空白组有明显差异?葛根素可以激活碱性磷酸酶活性,初级成骨细胞分化?葛根素可以促进钙的沉积,刺激骨形成?使用ERK1/2 阻断剂PD8089 后,或用抑制剂SB203580阻断p38 MAPK 信号通路后,细胞增殖?碱性磷酸酶含量以及钙沉积量均较葛根素组有所下降?葛根素组(T group)BMP-2 表达高于对照组( P <0. 05);葛根素+PD8089 组(T+PD group) 低于T 组高于对照组;葛根素+SB203580 组(T+SB group)钙沉积量较T 组明显降低( P <0. 05),与对照组BMP-2 表达量相比亦降低( P <0. 05)?结论 葛根素在骨细胞周期过程中,ERK1/2 和p38 MAPK 信号通路对骨分化和骨形成起到了调控作用?
英文摘要:
      Objective To explore the mechanism by which puerarin stimulates osteogenesis and bone formation through the ERK1/2 and p38 MAPK signaling pathways. Methods Adult osteoblasts (MC3T3-E1) cell culture was used in this study.The proliferative ability and the influence of growth curve of the cells after added different concentrations of puerarin were compared by MTT staining. The effect of puerarin on the differentiation of osteoblasts was assessed by measuring alkaline phosphatase activity and the effect of puerarin on bone formation was analyzed by measuring calcium deposition. Through the addition of ERK1/2 blocker PD8089 and p38 inhibitor SB203580, the mechanisms of involvement of the ERK1/2 and p38 MAPK signaling pathways in stimulating osteogenesis and bone formation were analyzed. Western blotting was used to detect the expression of bone morphogenetic protein-2 (BMP-2). Results Different concentrations of puerarin promoted the proliferation of osteoblasts to different degrees. The effect of puerarin at 0. 1 mol/ L was the most significant. The trend of proliferation was not pronounced on the first and third days compared with the level in the blank group, but significant differences emerged between the fifth and seventh days. Puerarin activated alkaline phosphatase activity and promoted the differentiation of primary osteoblasts. It also promoted calcium deposition and stimulated bone formation. After using the ERK1/2 blocker PD8089 or blocking the p38 MAPK signaling pathway with the inhibitor SB203580, cell proliferation, alkaline phosphatase content, and calcium deposition were lower than those of the puerarin group. The expression of BMP-2 in group puerarin (T) was higher than that of the control group ( P <0. 05), and that of group puerarin+PD 8089(T+PD)was lower than that of group T, while the amount of calcium deposition in the group puerarin+SB 203580(T+SB)was significantly lower than that in group T ( P <0. 05), and there was a decrease ( P <0. 05)in the BMP-2 expression compared with that in the control group. Conclusions In the cell cycle in bone, puerarin and the ERK1/2 and p38 MAPK signaling pathways play regulatory roles in bone differentiation and bone formation.
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