张山锋;马中希;叶志伟;刘珍星;杨钟华;孙志博;

• 1:武汉市第四医院华中科技大学同济医学院附属普爱医院骨科

摘要(Abstract)：

[目的]探讨缝隙连接蛋白43(Cx43)在淫羊藿苷(ICN)诱导人骨髓间充质干细胞(MSCs)成骨分化中的作用。[方法]密度梯度离心法体外分离培养MSCs,取第三代细胞实验,流式细胞仪测定细胞表面标记物。实验分四组:(1)空白对照组,(2)成骨诱导组,(3)1-庚醇组,(4)ICN组。MTT检测不同时间点2.5μmol/L 1-庚醇的细胞毒作用。分别用含0、10-9mol/L ICN、10-9mol/L ICN+2.5μmol/L 1-庚醇的成骨诱导液(ODM)诱导培养14 d后,实时定量PCR检测各组成骨相关基因及Cx43的表达情况,Western blot检测MSCs成骨分化过程中Cx43蛋白的表达情况,碱性磷酸酶(ALP)定量测定各组细胞ALP含量。[结果]细胞高表达CD13、CD44,低表达CD14、CD45。2.5μmol/L 1-庚醇对细胞没有明显的毒性抑制作用。ICN诱导14 d后I型胶原、ALP、骨钙素(OCN)、RUNX2及Cx43的表达显著增加。2.5μmol/L 1-庚醇干预后I型胶原、ALP、RUNX2及OCN表达明显减弱,但仍高于空白对照组及成骨诱导组,其中Cx43的表达无明显变化。ALP定量分析结果显示ICN能促进ALP合成与分泌,其含量显著高于其他三组,但ICN的成骨诱导作用部分被1-庚醇所阻断。[结论]Cx43介导的缝隙链接细胞间通讯在ICN诱导成骨过程中起着很重要作用。

关键词(KeyWords)： 成骨分化;淫羊藿苷;1-庚醇;缝隙连接蛋白43

Abstract:

Keywords:

基金项目(Foundation): 武汉市卫计委资助项目(No.WX16C28);; 国家自然科学基金资助项目(No.81501894)

作者(Author): 张山锋;马中希;叶志伟;刘珍星;杨钟华;孙志博;

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参考文献(References)：

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