IGFBP5 enhances osteogenic differentiation potential of periodontal ligament stem cells and Wharton's jelly umbilical cord stem cells, via the JNK and MEK/Erk signalling pathways
IGFBP5 enhances osteogenic differentiation potential of periodontal ligament stem cells and Wharton's jelly umbilical cord stem cells, via the JNK and MEK/Erk signalling pathways
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IGFBP5通过JNK和MEK/Erk信号通路增强牙周膜干细胞和沃顿胶脐带干细胞的成骨分化潜力
DOI：
10.1111/cpr.12284
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发表时间：
2016
影响因子：
8.5
通讯作者：
Fan Zhipeng
中科院分区：
文献类型：
--
作者：
Wang Yuejun;Jia Zhi;Diao Shu;Lin Xiao;Lian Xiaomeng;Wang Liping;Dong Rui;Liu Dayong;Fan Zhipeng
目的骨髓间充质干细胞(MSC)介导的组织再生是一种很有前途的组织修复策略，但其分子机制尚不清楚，限制了MSCs的应用。我们先前的研究表明，胰岛素样生长因子结合蛋白5(IGFbp5)对MSCs的成骨分化具有重要作用，但其定向分化的分子机制尚不清楚。在这项研究中，我们研究了Igfbp5在调节成骨分化潜能中的分子作用。材料和方法从牙周膜组织中分离出牙周膜干细胞。沃顿氏脐带干细胞凝胶(WJCMSCs)是商业化获得的。用慢病毒IGFBP5shRNA沉默IGFBP5。用表达野生型IGFBP5的逆转录病毒在WJCMSCs中过表达IGFBP5。用重组人Igfbp5蛋白(RhIGFBP5)处理PDLSCs 24小时，Western印迹分析MAPK信号转导通路，碱性磷酸酶(ALP)活性、茜素红染色和钙离子定量分析研究成骨分化潜能。结果IGFBP5或rhIGFBP5过表达均可使WJCMSCs和PDLSCs中磷酸化c-Jun氨基末端激酶(p-JNK)、磷酸化丝裂原活化蛋白激酶1和2(p-MEK1/2)和磷酸化细胞外调节蛋白激酶(p-ERK1/2)表达增加。沉默的IGFBP5可有效抑制PDLSCs和WJCMSCs中p-JNK、p-ERK1/2和p-MEK1/2的表达。结论IGFBP5通过JNK和MEK/Erk信号通路促进PDLSCs和WJCMSC的成骨分化潜能。
ObjectivesMesenchymal stem cell (MSC)‐mediated tissue regeneration represents a promising strategy for repair of tissue defects, but its molecular mechanisms remain unclear, restricting the use of MSCs. Our previous study indicated that insulin‐like growth factor‐binding protein 5 (IGFBP5) exerted a valuable effect on osteogenic differentiation of MSCs, but its molecular mechanisms underlying directed differentiation remained unclear. In this study, we have investigated the molecular role of IGFBP5 in regulating this osteogenic differentiation potential.Materials and methodsPeriodontal ligament stem cells (PDLSCs) were isolated from periodontal ligament tissue. Wharton's jelly of umbilical cord stem cells (WJCMSCs) was obtained commercially. LentiviralIGFBP5shRNA was used to silenceIGFBP5. Retroviruses expressing wild‐typeIGFBP5were used to overexpressIGFBP5in the WJCMSCs. Recombinant human IGFBP5 protein (rhIGFBP5) was used to treat PDLSCs for 24 h. Western blot analysis was used to detect the MAPK signalling pathway, and alkaline phosphatase (ALP) activity, Alizarin Red staining and quantitative calcium analysis were used to study osteogenic differentiation potentials.ResultsOverexpression ofIGFBP5or rhIGFBP5 increased expression levels of phosphorylated c‐Jun N‐terminal kinase (p‐JNK), phosphorylated mitogen‐activated protein kinase 1 and 2 (p‐MEK1/2) and phosphorylated extracellular regulated protein kinases (p‐Erk1/2) in both WJCMSCs and PDLSCs. Consistently, silencedIGFBP5was found to effectively inhibit expression of p‐JNK, p‐Erk1/2 and p‐MEK1/2 in PDLSCs and WJCMSCs. Furthermore, inhibition of JNK by its inhibitor, SP600125, or MEK/Erk signalling by its inhibitor, PD98059, dramatically blockedIGFBP5‐enhanced ALP activity and in vitro mineralization in both PDLSCs and WJCMSCs.ConclusionsOur results demonstrated thatIGFBP5promoted osteogenic differentiation potentials of PDLSCs and WJCMSCsviathe JNK and MEK/Erk signalling pathways.