Abnormal osteogenic and chondrogenic differentiation of human mesenchymal stem cells from patients with adolescent idiopathic scoliosis in response to melatonin.
Abnormal osteogenic and chondrogenic differentiation of human mesenchymal stem cells from patients with adolescent idiopathic scoliosis in response to melatonin.
复制标题
青少年特发性脊柱侧凸患者的人间充质干细胞对褪黑素的反应异常成骨和软骨分化
DOI：
10.3892/mmr.2016.5384
复制
发表时间：
2016-08
影响因子：
3.4
通讯作者：
Su P
中科院分区：
文献类型：
--
作者：
Chen C;Xu C;Zhou T;Gao B;Zhou H;Chen C;Zhang C;Huang D;Su P
青少年特发性脊柱侧凸（AIS）患者的膜性和软骨内成骨异常尚不完全清楚。为了研究AIS中褪黑激素信号通路的异常和细胞对褪黑激素的反应，使用人间充质干细胞（hMSCs）进行了成骨和软骨分化的病例对照研究。通过体格检查和影像学检查诊断AIS。分别从AIS患者和对照组（n=12）骨髓中分离hMSCs，采用密度梯度离心纯化。western blotting检测褪黑激素受体（MTs） 1和2的表达水平。通过在含有载体或50 nM褪黑素的成骨和软骨培养基中培养hMSCs诱导成骨和软骨分化。进行碱性磷酸酶（ALP）活性测定、糖胺聚糖（GAG）定量分析、逆转录-定量聚合酶链反应分析。与对照组相比，AIS组MT2表达较低。褪黑素增加了正常对照hMSCs中ALP活性、GAG合成，上调了成骨和软骨分化相关基因的表达，包括ALP、骨桥蛋白、骨钙素、矮子相关转录因子2、II型胶原、X型胶原、聚集蛋白和性别决定区Y-box 9，但对AIS组没有影响。因此，AIS hMSCs在成骨和软骨分化过程中表现出对褪黑激素的异常细胞反应，这可能与异常的膜性和软骨内成骨以及骨骼生长有关。这些结果表明褪黑素通过MT2受体对AIS患者异常成骨和软骨分化具有潜在的调节作用。
Abnormalities of membranous and endochondral ossification in patients with adolescent idiopathic scoliosis (AIS) remain incompletely understood. To investigate abnormalities in the melatonin signaling pathway and cellular response to melatonin in AIS, a case-control study of osteogenic and chondrogenic differentiation was performed using human mesenchymal stem cells (hMSCs). AIS was diagnosed by physical and radiographic examination. hMSCs were isolated from the bone marrow of patients with AIS and control subjects (n=12 each), and purified by density gradient centrifugation. The expression levels of melatonin receptors (MTs) 1 and 2 were detected by western blotting. Osteogenic and chondrogenic differentiation was induced by culturing hMSCs in osteogenic and chondrogenic media containing vehicle or 50 nM melatonin. Alkaline phosphatase (ALP) activity assays, quantitative glycosaminoglycan (GAG) analysis, and reverse transcription-quantitative polymerase chain reaction analysis were performed. Compared with controls, MT2 demonstrated low expression in the AIS group. Melatonin increased ALP activity, GAG synthesis and upregulated the expression of genes involved in osteogenic and chondrogenic differentiation including, ALP, osteopontin, osteocalcin, runt-related transcription factor 2, collagen type II, collagen type X, aggrecan and sex-determining region Y-box 9 in the normal control hMSCs, but did not affect the AIS groups. Thus, AIS hMSCs exhibit abnormal cellular responses to melatonin during osteogenic and chondrogenic differentiation, which may be associated with abnormal membranous and endochondral ossification, and skeletal growth. These results indicate a potential modulating role of melatonin via the MT2 receptor on abnormal osteogenic and chondrogenic differentiaation in patients with AIS.