PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway.
PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway.
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DOI：
10.1590/1414-431x20165988
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发表时间：
2017-02-16
期刊：
影响因子：
--
通讯作者：
Qiao YJ
中科院分区：
文献类型：
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作者：
Wang H;Shan XB;Qiao YJ
本研究旨在阐明丙酮酸脱氢酶激酶异构体2 （PDK2）在间充质干细胞（MSCs）软骨分化中的作用和机制。从体重300-400 g的Sprague-Dawley大鼠（雌性5只，雄性5只）股骨和胫骨中分离MSCs。将PDK2过表达和敲低转染到MSCs中，然后评估细胞活力、粘附和迁移。此外，通过定量逆转录聚合酶链式反应（qRT-PCR）检测了异常PDK2在软骨形成标志物sry相关高迁移率群盒6 （Sox6）、ΙΙ型前胶原基因（COL2A1）、软骨寡聚基质蛋白（COMP）、聚集蛋白（AGC1）、ΙX型前胶原基因（COL9A2）和胶原1型α 1 （COL1A1）中的作用。检测c-Jun n末端激酶（JNK）、p38丝裂原活化蛋白激酶（MAPK）和细胞外调节蛋白激酶（ERK）的表达。过表达PDK2可促进间充质干细胞的细胞活力、粘附并抑制细胞迁移（均P<0.05）。qRT-PCR分析显示，PDK2过表达后，所有软骨形成标志物的mRNA表达均显著增加（P<0.01或P<0.05）。与对照组相比，PDK2过表达还导致MSCs中JNK、p38MAPK和ERK的mRNA和蛋白表达显著增加（P<0.01或P<0.05）。同时，沉默PDK2对间充质干细胞产生相反的作用。本研究初步揭示了PDK2在骨髓间充质干细胞成软骨分化中的积极作用及其潜在机制。为揭示PDK2的功能奠定了理论基础，为骨关节炎软骨病变的修复提供了良好的基础。
This study was undertaken to clarify the role and mechanism of pyruvate dehydrogenase kinase isoform 2 (PDK2) in chondrogenic differentiation of mesenchymal stem cells (MSCs). MSCs were isolated from femurs and tibias of Sprague-Dawley rats, weighing 300-400 g (5 females and 5 males). Overexpression and knockdown of PDK2 were transfected into MSCs and then cell viability, adhesion and migration were assessed. Additionally, the roles of aberrant PDK2 in chondrogenesis markers SRY-related high mobility group-box 6 (Sox6), type ΙΙ procollagen gene (COL2A1), cartilage oligomeric matrix protein (COMP), aggrecan (AGC1), type ΙX procollagen gene (COL9A2) and collagen type 1 alpha 1 (COL1A1) were measured by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The expressions of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK) and extracellular regulated protein kinase (ERK) were measured. Overexpressing PDK2 promoted cell viability, adhesion and inhibited cell migration in MSCs (all P<0.05). qRT-PCR assay showed a potent increase in the mRNA expressions of all chondrogenesis markers in response to overexpressing PDK2 (P<0.01 or P<0.05). PDK2 overexpression also induced a significant accumulation in mRNA and protein expressions of JNK, p38MAPK and ERK in MSCs compared to the control (P<0.01 or P<0.05). Meanwhile, silencing PDK2 exerted the opposite effects on MSCs. This study shows a preliminary positive role and potential mechanisms of PDK2 in chondrogenic differentiation of MSCs. It lays the theoretical groundwork for uncovering the functions of PDK2 and provides a promising basis for repairing cartilage lesions in osteoarthritis.