| 摘要: |
| 为探究猪（Sus scrofa）骨髓间充质干细胞（Bone marrow mesenchymal stem cells， PBMSCs）成软骨分化过程中能量代谢动态变化以及与软骨形成之间的关系，本研究分离并鉴定猪骨髓间充质干细胞，并通过建立诱导分化培养体系获得猪软骨细胞。对PBMSCs成软骨诱导分化第1、6、12和18天的细胞进行软骨形成标记基因、能量代谢相关基因表达模式分析以及能量代谢产物含量检测，且对软骨形成标记基因与能量代谢产物进行相关性分析。结果表明：1）在成软骨分化过程，软骨形成标记基因II型胶原α1链（Collagen type II alpha 1 chain，COL2A1）、SRY-box转录因子9（SRY-box transcription factor 9，SOX9）、聚集蛋白聚糖（Aggrecan，ACAN）、基质金属蛋白酶13（Matrix metallopeptidase 13，MMP13）、RUNX家族转录因子2（RUNX family transcription factor 2，RUNX2）表达水平显著提高（P<0.05）；糖酵解限速酶基因己糖激酶1（Hexokinase 1，HK1）、丙酮酸激酶L/R（Pyruvate kinase L/R，PKLR）和乳酸转运关键基因溶质载体家族16成员1（Solute carrier family 16 member 1，SLC16A1）表达水平显著提高（P<0.05），葡萄糖转运关键基因溶质载体家族2成员4（Solute carrier family 2 member 4，SLC2A4）表达水平显著降低（P<0.05）；2）随着成软骨分化时间增加，胞内ATP含量和葡萄糖含量显著降低（P<0.05），胞内乳酸、乳酸和葡萄糖比率显著增加（P<0.05）。3）成软骨分化关键基因与能量代谢产物的相关性分析表明，PBMSCs成软骨过程与糖酵解代谢显著相关（P<0.05）。综上，本研究成功建立了一种诱导PBMSCs分化成软骨细胞的方法，并发现在间充质干细胞诱导成软骨细胞过程中，细胞氧化磷酸化水平逐渐降低，糖酵解代谢逐渐增强，为后续深入解析猪体长发育机制提供新视角。 |
| 关键词: 猪 骨髓间充质干细胞 细胞分化 软骨细胞 能量代谢 |
| DOI：10.11841/j.issn.1007-4333.2024.02.14 |
| 投稿时间：2023-06-16 |
| 基金项目:三亚崖州湾科技城管理局2020年度科技计划项目（SKJC-2020-02-007）；国家自然科学基金项目（32072700；U22A20508）；国家重点研发计划（2022YFF1003401）；山东省重点研发计划农业良种工程（2022LZGQY007）；财政部和农业农村部国家现代农业产业技术体系（CARS-35） |
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| Analysis of dynamic changes in energy metabolism during chondrogenic differentiation of porcine bone marrow mesenchymal stem cells |
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XUE Mingming1, LI Fengmiao1, LUO Yabiao1, YANG Xiaoyang1, XIE Fuyin1, CHAN Shuheng1, TANG Qiguo1, FANG Meiying1,2*
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| (1.College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;2.Sanya Research Institute, China Agricultural University, Sanya 572025, China) |
| Abstract: |
| In order to explore the dynamic changes in energy metabolism during the chondrogenic differentiation of porcine bone marrow mesenchymal stem cells （PBMSCs） and its relationship with cartilage formation， PBMSCs were isolated and identified， and porcine chondrocytes were obtained though an induction differentiation culture system . Analyses of the expression patterns of cartilage marker genes and energy metabolism-related genes were performed， as well as measurements of energy metabolites content， in PBMSCs induced to differentiate into chondrocytes for 1， 6， 12， and 18 days. Pearson correlation analyses between cartilage marker genes and energy metabolism products were conducted. The results showed that： 1） During the chondrogenic differentiation of PBMSCs， the expression levels of cartilage marker genes， including COL2A1， SOX9， ACAN， MMP13， and RUNX2， were significantly increased （P<0.05）. The expression levels of the glycolytic rate-limiting enzyme genes HK1， PKLR， and the lactate transporter key gene SLC16A1 were significantly upregulated （P<0.05）， while the expression level of the glucose transport critical gene， SLC2A4， was significantly reduced （P<0.05）. 2） With the increase of chondrogenic differentiation time， intracellular levels of ATP and glucose were significantly decreased （P<0.05）， while the intracellular ratio of lactate to glucose was significantly increased （P<0.05）. 3） The correlation analysis of key chondrogenic genes and energy metabolic products suggested a significant association between the chondrogenesis process of PBMSCs and glycolytic metabolism （P<0.05）. In summary， this study successfully developed a method of inducing PBMSCs into chondrocyte， and revealed oxidative phosphorylation levels gradually decreased， and glycolytic metabolism gradually increased during the PBMSC-induced chondrogenic differentiation process. These findings provide a new perspective for further analysis of pig body length development. |
| Key words: pig bone marrow mesenchymal stem cells cell differentiation chondrocyte energy metabolism |
