关于HMGB1促进骨髓间充质干细胞成骨分化及迁移能力的分子机制研究-外科学（骨科学）专业论文.docxVIP

浙江大学硕士学位论文 浙江大学硕士学位论文 中文摘要 关于HMGBl促进骨髓间充质于细胞成骨分化 及迁移能力的分子机制研究 中文摘要 众所周知，骨髓间充质干细胞(MSC)的迁移和成骨分化能力在骨折愈合中起 到至关重要的作用。High mobility group box 1(HMGBl)作为在骨折部位广泛存在 的一种炎症因子，它对于MSC的化学刺激作用已被证实。在研究中，为较全面的 揭示HMGBl对MSC成骨分化和迁移的影响以及其分子机制，我们将检测在 HMGBl的刺激下MSC生物功能的改变及细胞因子的合成及相关信号通路的激 活。 实验中，我们通过茜素红染色及碱性磷酸酶染色检测了MSC的成骨分化水平， 通过划痕实验测试了MSC的迁移能力。同时为了检测MSC合成的细胞因子，我 们使用了抗体芯片技术。检测结果表明，HMGBl刺激MSC合成了多种细胞因子， 而这些细胞因子激活了细胞的相应的信号通路。后续我们通过westem-blot技术配 合信号通路抑制剂的使用，进一步论证了细胞通路的激活及其对MSC的影响。 结果表明，HMGBl刺激了MSC的细胞因子的合成。这些细胞因子通过激活 了MSC的信号通路。在所激活的信号通路的调控下，MSC的成骨分化能力及迁 移能力得到显著的增强。 关键词：骨髓间充质干细胞，成骨分化，迁移，细胞因子，信号通路 万方数据 The The molecular mechanism involved in the effects of HMGBl on mesenchymal stem cell osteoblastic differentiation and migration Abstract The migration of mesenchymal stem cells(MSC)and osteogenic differentiation play an important role in fracture healing．High mobility group box 1(HMGBl)，a widely distributed inflammatory factor in fractures，has been confirmed to act as a chemoattractant to MSC．To investigate the effect of HMGB 1 on MSC osteogenic differentiation and migration and the underlying mechanism，we evaluated the synthesis of MSC chemokines and the consequent activation of signaling pathways following HMGB 1 stimulation． We use Alizarin red staining and ALP staining to prove the osteogenic differentiation．And use test the migration by scratch assay．To detect cytokine secretion， antibody array assays were performed．These results suggest that HMGB 1 enhances the secretion of multiple cytokines by MSC and promotes biological functions through the signaling pathway．Then we use the western-blot and inhibitor to test the function of signaling pathway more deeply． These results suggest that HMGB 1 induce the cytokine secretion of MSC．And the cytokines promote the signaling pathway．At last，the biological functions of MSC has been promoted through the signaling pathway．This study may provide a theoretical basis for the development of new techniques for the treatment of fractures in the future．