Melatonin enhances chondrogenic differentiation of human mesenchymal stem cells
Melatonin enhances chondrogenic differentiation of human mesenchymal stem cells
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褪黑素增强人间充质干细胞的软骨分化
DOI：
10.1111/jpi.12098
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发表时间：
2014-01-01
影响因子：
10.3
通讯作者：
Su, Peiqiang
中科院分区：
文献类型：
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作者：
Gao, Wenjie;Lin, Mianlong;Su, Peiqiang
膜内骨化和软骨内骨化是骨形成和骨折愈合的两种途径。越来越多的证据表明，褪黑素影响成骨细胞分化，但对褪黑素在软骨细胞分化过程中的作用知之甚少。本研究探讨褪黑素对人骨髓间充质干细胞(MSCs)向软骨细胞分化的影响。在含有载体或50 nm褪黑素的软骨细胞培养液中进行高密度微团培养，诱导细胞向软骨方向分化。组织学观察和糖胺多聚糖(GAG)定量分析显示，褪黑素组软骨组织体积较大，GAG、II型和X型胶原含量均高于未处理组。实时定量RT-PCR分析表明，褪黑素处理显著上调了与软骨分化相关的基因的表达，包括聚集素(Acan)、II型胶原(COL2A1)、X型胶原(COL10A1)、性别决定区SRY-box 9(SOX9)、矮小相关转录因子2(RUNX2)以及软骨分化的有效诱导剂骨形态发生蛋白2(BMP2)。免疫荧光染色检测软骨诱导MSCs中褪黑素膜受体(MT)MT1和MT2的表达。褪黑素受体拮抗剂Luzindole可部分阻断褪黑素增加软骨组织大小和GAG合成的能力，并在分化7d后完全逆转褪黑素对Acan、COL2A1、COL10A1、SOX9和BMP2基因表达的影响。这些发现表明，褪黑素至少部分地通过褪黑素受体促进了人MSCs的软骨分化。
Intramembranous ossification and endochondral ossification are two ways through which bone formation and fracture healing occur. Accumulating amounts of evidence suggests that melatonin affects osteoblast differentiation, but little is known about the effects of melatonin on the process of chondrogenic differentiation. In this study, the effects of melatonin on human mesenchymal stem cells (MSCs) undergoing chondrogenic differentiation were investigated. Cells were induced along chondrogenic differentiation via high‐density micromass culture in chondrogenic medium containing vehicle or 50 nm melatonin. Histological study and quantitative analysis of glycosaminoglycan (GAG) showed induced cartilage tissues to be larger and richer in GAG, collagen type II and collagen type X in the melatonin group than in the untreated controls. Real‐time RT‐PCR analysis demonstrated that melatonin treatment significantly up‐regulated the expression of the genes involved in chondrogenic differentiation, including aggrecan (ACAN), collagen type II (COL2A1), collagen type X (COL10A1), SRY (sex‐determining region Y)‐box 9 (SOX9), runt‐related transcription factor 2 (RUNX2) and the potent inducer of chondrogenic differentiation, bone morphogenetic protein 2 (BMP2). And the expression of melatonin membrane receptors (MT) MT1 and MT2 were detected in the chondrogenic‐induced‐MSCs by immunofluorescence staining. Luzindole, a melatonin receptor antagonist, was found to partially block the ability of melatonin to increase the size and GAG synthesis of the induced cartilage tissues, as well as to completely reverse the effect of melatonin on the gene expression of ACAN, COL2A1, COL10A1, SOX9 and BMP2 after 7 days of differentiation. These findings demonstrate that melatonin enhances chondrogenic differentiation of human MSCs at least partially through melatonin receptors.