Background Bone morphogenetic protein 9 （BMP9） and Wnt/13-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells （MSCs）, but the role of Wnt/13-catenin signaling pathway...Background Bone morphogenetic protein 9 （BMP9） and Wnt/13-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells （MSCs）, but the role of Wnt/13-catenin signaling pathway in BMP9-induced osteogenic differentiation is not well understood. Thus, our experiment was undertaken to investigate the interaction between BMP9 and Wnt/13-catenin pathway in inducing osteogenic differentiation of MSCs. Methods C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP9, Wnt3a, and BMPg＋Wnt3a. ALP, the early osteogenic marker, was detected by quantitative and staining assay. Later osteogenic marker, mineral calcium deposition, was determined by Alizarin Red S staining. The expression of osteopotin （OPN）, osteocalcin （OC）, and Runx2 was analyzed by Real time PCR and Western blotting. In vivo animal experiment was carried out to further confirm the role of Wnt3a in ectopic bone formation induced by BMP9. Results The results showed that Wnt3a enhanced the ALP activity induced by BMP9 and increased the expressions of OC and OPN, with increase of mineral calcium deposition in vitro and ectopic bone formation in vivo. Furthermore, we also found that Wnt3a increased the level of Runx2, an important nuclear transcription factor of BMP9. Conclusion Canonical Wnt/13-catenin signal pathway may play an important role in BMP9-induced osteogenic differentiation of MSCs, and Runx2 may be a linkage between the two signal pathways.展开更多
背景:骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有很强的促进干细胞成骨分化的潜能。大量研究表明,BMP9可在动物体内促进骨缺损的修复。目的:探究BMP9调控干细胞分化的机制,讨论影响BMP9促进干细胞成骨分化的潜在因素以及B...背景:骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有很强的促进干细胞成骨分化的潜能。大量研究表明,BMP9可在动物体内促进骨缺损的修复。目的:探究BMP9调控干细胞分化的机制,讨论影响BMP9促进干细胞成骨分化的潜在因素以及BMP9在骨组织工程中的应用前景。方法:在PubMed、Web of Science、万方、维普、CNKI数据库中以“bone morphogenetic protein 9/BMP9,mesenchymal stem cell,stem cell,bone regeneration,bone reconstruction”“骨形态发生蛋白9,间充质干细胞,干细胞,骨再生,骨重建”为关键词检索1990至2020年的相关文献,排除重复、与研究目的无关或低质量的文献,共纳入文献67篇进行总结分析。结果与结论:BMP9作为骨形态发生蛋白家族的关键分子,具有极强的促进干细胞分化与骨组织再生的能力。BMP9主要通过BMP9-SMAD信号通路和BMP9-MAPK信号通路来实现对干细胞分化的调控作用。TGF-β、Wnt/β-catenin和Notch信号通路可以影响BMP9信号通路关键分子的表达,间接影响BMP9的成骨分化作用。在口腔颌面部骨组织工程与骨关节疾病模型中,BMP9可以显著促进骨组织再生、骨缺损的修复与骨组织的重建。现有研究仍存在一定局限性,包括BMP9调控干细胞分化与骨组织再生的具体机制仍有待进一步探讨,BMP9对其他组织器官的毒副作用仍有待评估等。展开更多
前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有较强的诱导间充质干细胞成骨分化的能力.为进一步揭示其诱导和调控间充质干细胞成骨分化的机理,利用BMP9重组腺病毒感染间充质干细胞C3H10T1/2,通过体外细胞实验和...前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有较强的诱导间充质干细胞成骨分化的能力.为进一步揭示其诱导和调控间充质干细胞成骨分化的机理,利用BMP9重组腺病毒感染间充质干细胞C3H10T1/2,通过体外细胞实验和体内动物实验,初步分析BMP9是否可通过p38激酶途径调控间充质干细胞成骨分化.结果发现,BMP9可以通过促进p38激酶磷酸化而导致其活化,p38抑制剂SB203580可抑制由BMP9诱导的C3H10T1/2细胞的碱性磷酸酶(alkalinephosphatase,ALP)活性、骨桥蛋白(osteopontin,OPN)表达和钙盐沉积,而且利用抑制剂SB203580抑制p38激酶活性后,BMP9诱导的Smad经典途径的激活也相应受到抑制,RNA干扰导致p38基因沉默同样也可抑制BMP9诱导的ALP活性、OPN表达、钙盐沉积以及裸鼠皮下异位成骨.因此,BMP9可通过活化p38激酶途径调控间充质干细胞C3H10T1/2成骨分化.展开更多
基金This work was supported by a grant from the National Natural Science Foundation of China （No. 31070875）.
文摘Background Bone morphogenetic protein 9 （BMP9） and Wnt/13-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells （MSCs）, but the role of Wnt/13-catenin signaling pathway in BMP9-induced osteogenic differentiation is not well understood. Thus, our experiment was undertaken to investigate the interaction between BMP9 and Wnt/13-catenin pathway in inducing osteogenic differentiation of MSCs. Methods C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP9, Wnt3a, and BMPg＋Wnt3a. ALP, the early osteogenic marker, was detected by quantitative and staining assay. Later osteogenic marker, mineral calcium deposition, was determined by Alizarin Red S staining. The expression of osteopotin （OPN）, osteocalcin （OC）, and Runx2 was analyzed by Real time PCR and Western blotting. In vivo animal experiment was carried out to further confirm the role of Wnt3a in ectopic bone formation induced by BMP9. Results The results showed that Wnt3a enhanced the ALP activity induced by BMP9 and increased the expressions of OC and OPN, with increase of mineral calcium deposition in vitro and ectopic bone formation in vivo. Furthermore, we also found that Wnt3a increased the level of Runx2, an important nuclear transcription factor of BMP9. Conclusion Canonical Wnt/13-catenin signal pathway may play an important role in BMP9-induced osteogenic differentiation of MSCs, and Runx2 may be a linkage between the two signal pathways.
文摘背景:骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有很强的促进干细胞成骨分化的潜能。大量研究表明,BMP9可在动物体内促进骨缺损的修复。目的:探究BMP9调控干细胞分化的机制,讨论影响BMP9促进干细胞成骨分化的潜在因素以及BMP9在骨组织工程中的应用前景。方法:在PubMed、Web of Science、万方、维普、CNKI数据库中以“bone morphogenetic protein 9/BMP9,mesenchymal stem cell,stem cell,bone regeneration,bone reconstruction”“骨形态发生蛋白9,间充质干细胞,干细胞,骨再生,骨重建”为关键词检索1990至2020年的相关文献,排除重复、与研究目的无关或低质量的文献,共纳入文献67篇进行总结分析。结果与结论:BMP9作为骨形态发生蛋白家族的关键分子,具有极强的促进干细胞分化与骨组织再生的能力。BMP9主要通过BMP9-SMAD信号通路和BMP9-MAPK信号通路来实现对干细胞分化的调控作用。TGF-β、Wnt/β-catenin和Notch信号通路可以影响BMP9信号通路关键分子的表达,间接影响BMP9的成骨分化作用。在口腔颌面部骨组织工程与骨关节疾病模型中,BMP9可以显著促进骨组织再生、骨缺损的修复与骨组织的重建。现有研究仍存在一定局限性,包括BMP9调控干细胞分化与骨组织再生的具体机制仍有待进一步探讨,BMP9对其他组织器官的毒副作用仍有待评估等。
文摘前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有较强的诱导间充质干细胞成骨分化的能力.为进一步揭示其诱导和调控间充质干细胞成骨分化的机理,利用BMP9重组腺病毒感染间充质干细胞C3H10T1/2,通过体外细胞实验和体内动物实验,初步分析BMP9是否可通过p38激酶途径调控间充质干细胞成骨分化.结果发现,BMP9可以通过促进p38激酶磷酸化而导致其活化,p38抑制剂SB203580可抑制由BMP9诱导的C3H10T1/2细胞的碱性磷酸酶(alkalinephosphatase,ALP)活性、骨桥蛋白(osteopontin,OPN)表达和钙盐沉积,而且利用抑制剂SB203580抑制p38激酶活性后,BMP9诱导的Smad经典途径的激活也相应受到抑制,RNA干扰导致p38基因沉默同样也可抑制BMP9诱导的ALP活性、OPN表达、钙盐沉积以及裸鼠皮下异位成骨.因此,BMP9可通过活化p38激酶途径调控间充质干细胞C3H10T1/2成骨分化.
