HSP70促进骨髓间充质干细胞成骨分化的实验研究

发布时间：2018-06-05 17:40

本文选题：热休克蛋白70 + 人骨髓间充质干细胞　；参考：《浙江大学》2016年硕士论文

【摘要】：目的：研究HSP70对骨髓间充质干细胞成骨分化的影响及其相关机制方法：在成骨分化培养基中添加不同浓度热休克蛋白70(HSP70)对人骨髓间充质干细胞(hMSC)进行培养。采用碱性磷酸酶(ALP)活力检测以及茜素红染色了解HSP70在成骨分化早期和晚期的影响。采用PCR及WB技术检测成骨相关标志物的表达变化。最后通过基因芯片技术和通路富集分析研究HSP70促进成骨分化相通路。结果：实验结果显示HSP70可显著提高ALP活力,同时促进钙结节的形成,且存在时间依赖性和剂量依赖性。PCR结果表明HSP70可显著促进成骨相关基因RUNX2,OSX和ALP的表达,WB结果显示HSP70可显著促进成骨相关蛋白SP7,ALP和BMP2的表达。基因芯片和通路富集分析提示ERK信号通路与HSP70促进hMSC的成骨分化关系密切。结论：HSP70通过激活ERK信号通路从而促进hMSC的成骨分化。
[Abstract]:Aim: to study the effect of HSP70 on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and its related mechanisms: human bone marrow mesenchymal stem cells (hMSCs) were cultured with HSP70 in different concentrations of heat shock protein 70 (HSP70). Alkaline phosphatase (ALP) activity and alizarin red staining were used to investigate the effect of HSP70 on early and late osteogenic differentiation. Polymerase chain reaction (PCR) and WB technique were used to detect the expression of osteoblast related markers. Finally, gene chip technology and pathway enrichment analysis were used to study the pathway of HSP70 promoting osteogenic differentiation. Results: HSP70 significantly increased the activity of ALP and promoted the formation of calcium nodules. The results of time-dependent and dose-dependent .PCR showed that HSP70 could significantly promote the expression of osteoblast-associated genes RUNX2OSX and ALP. The results showed that HSP70 could significantly promote the expression of osteoblast-associated proteins SP7, ALP and BMP2. Gene chip and pathway enrichment analysis suggested that ERK signaling pathway was closely related to HSP70 promoting osteogenic differentiation of hMSC. Conclusion: HSP70 promotes osteogenic differentiation of hMSC by activating ERK signaling pathway.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R683

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