【摘要】 目的探讨Talin1在骨髓间充质干细胞(Mesenchymal stem cell,MSC)成骨分化中的作用及其调控机制。方法诱导MSC成骨分化后通过qRT-PCR及Western blot检测TLN1的表达变化,在MSC中敲低TLN1后采用CCK-8检测细胞增殖能力改变,利用ALP活性检测及茜素红染色观察成骨分化变化情况。通过Western blot实验研究TLN1在MSC成骨分化过程中参与的分子机制。结果 qPCR及Western blot结果提示,TLN1在成骨分化过程中表达上升;siRNA敲低TLN1表达后,MSC增殖无明显差异,ALP活性及矿化结节定量较对照组下降(P<0.05)。Western blot结果显示敲低TLN1表达后p-AKT表达下降,回补实验证明激活PI3K-AKT表达可回复MSC的成骨分化能力。结论诱导MSC成骨分化后TLN1表达逐渐上调,敲低TLN1后通过PI3K-AKT通路抑制骨髓间充质干细胞成骨分化。更多还原

【Abstract】 Objective To investigate the role and regulatory mechanism of Talin 1 in osteogenic differentiation of bone marrow mesenchymal stem cells. Methods The expression level of TLN1 in osteogenic differentiation of mesenchymal stem cells was detected by qRT-PCR and western blot. The proliferation of MSC was measured by CCK-8 assay after TLN1-siRNA intervention. The osteogenic differentiation capacity was determined through alizarin red S and alkaline phosphatase assays. Western blotting was used to study the molecular mechanism of TLN1 in the osteogenic differentiation of MSC.Results qRT-PCR and Western blot results suggested that TLN1 expression increased during osteogenic differentiation. The osteogenic differentiation capacity was decrease compared with the negative group after siRNA transfection. After transfection with siRNA,the expression of p-AKT was decreased by Western blot. Furthermore,the rescue experiment showed that active PI3 K/AKT pathway can reserve osteogenic differentiation capacity. Conclusion Talin1 expression was gradually up-regulated after induced osteogenic differentiation of MSC. TLN1 knockdown inhibited osteogenic differentiation of MSC through PI3 K/AKT pathway.更多还原