【摘要】 目的 研究机械刺激联合铁治疗通过TGFβ/Wnt信号通路介导细胞增殖促进骨愈合的潜在机制。方法 将18只10周龄雄性Balb/c小鼠随机分成正常组、骨折组、机械刺激/铁组，每组6只。制作小鼠胫骨骨折动物模型，酶联免疫吸附试验检测骨钙素、碱性磷酸酶、Runx2浓度，组织学分析骨组织体积，成骨细胞培养检测成骨细胞活力，Western Blot检测TGFβ/Wnt信号通路相关蛋白水平。结果 机械刺激/铁组骨钙素、碱性磷酸酶、Runx2浓度较骨折组高，骨折组较正常组高。Western Blotting检测机械刺激/铁组上调COL2A1蛋白表达水平，下调COL10A1和SOX9蛋白表达水平。与正常组相比，骨折组增殖细胞核抗原和Ki67蛋白水平低，机械刺激/铁组增殖细胞核抗原和Ki67蛋白水平较骨折组高。RT-PCR结果显示，与正常组相比，骨折组TGFβ、GSK3β和β-catenin mRNA水平上调，机械刺激/铁组mRNA水平上调被抑制，各组smad3 mRNA无明显变化。机械刺激/铁组成骨细胞COL2A1蛋白水平上调，TGFβ和β-catenin蛋白水平下调，促进成骨活性。结论 机械刺激联合铁治疗通过TGFβ/Wnt信号通路介导细胞增殖修复骨损伤和促进骨愈合。更多还原

【Abstract】 Objective To stady the potential mechanism of mechanical stimulation combined with iron therapy to promote bone healing through TGFβ/Wnt signaling pathway mediated cell proliferation.Methods Eighteen 10-week-old male Balb/c mice were randomly divided into normal group,fracture group and mechanical stimulation/iron group,with 6 mice in each group.The mouse tibia fracture animal model was made,and the concentrations of osteocalcin,alkaline phosphatase and Runx2were detected by ELISA,the bone tissue volume was analyzed by histology,osteoblast activity was detected by osteoblast culture,and the levels of TGFβ/Wnt signaling pathway-related proteins were detected by Western Blot.Results The concentrations of osteocalcin,alkaline phosphatase and Runx2 in mechanical stimulation/iron group were higher than those in fracture group,while those in fracture group were higher than those in normal group.Western Blotting detected up-regulated COL2A1protein expression and down-regulated COL10A1 and SOX9 protein expression in mechanical stimulation/iron group.Compared with the normal group,the levels of proliferating nuclear antigen and Ki67 protein were lower in the fracture group,while the levels of proliferating nuclear antigen and Ki67 protein were higher in the mechanical stimulation/iron group.RT-PCR detection results showed that compared with the normal group,the mRNA levels of TGFβ,GSK3β and β-catenin in the fracture group were up-regulated,while the mRNA levels of the mechanical stimulation/iron group were suppressed,and there was no significant change in smad3 mRNA in each group.The protein level of COL2A1 was up-regulated and the protein levels of TGFβ and β-catenin were down-regulated in mechanical stimulation/iron constitutive bone cells,which promoted osteogenic activity.Conclusion Mechanical stimulation combined with iron therapy mediates cell proliferation through TGFβ/Wnt signaling pathway to repair bone injury and promote bone healing.更多还原