青少年特发性脊柱侧凸患者骨髓间充质干细胞褪黑素受体表达的研究

目的:比较体外培养的青少年特发性脊柱侧凸(AIS)患者组和正常对照组骨髓间充质干细胞(BMSCs)褪黑素受体表达的差异性,并探讨其意义。方法:分别从患者髂前上棘处穿刺抽取10ml骨髓,肝素抗凝。采用密度梯度离心法分离BMSCs,体外培养并传至P3代,进行表型鉴定。抽提细胞的总RNA,采用RT-PCR方法检测2组褪黑素受体MT1、MT2 mRNA的表达水平。结果:分离的单个核细胞经培养至P3代,使用流式细胞仪进行细胞表型鉴定,体外培养细胞的表型与BMSCs的表面标志相符。AIS组MT1、MT2 mRNA的表达水平与正常对照组相比无统计学差异(P>0.05)。结论:AIS患者BMSCs褪黑素受体MT1、MT2的表达无异常。在干细胞水平,褪黑素受体与AIS的发病可能没有明显的相关性。

Studies of the expression of melatonin receptors(MT1,MT2) in BMSCs from adolescent idiopathic scoliosis patients

Objective:To investigate the expression of melatonin receptors(MT1,MT2) in BMSCs from adolescent idiopathic scoliosis patients.Methods:Twenty-eight volunteers were divided into two groups:AIS group consisted of 18 patients and control group consisted of 10 subjects.The human bone marrow anticoagulated by heparin was obtained from anterior superior iliac spine,and the BMSCs were isolated by density gradient centrifuge from the mononuclear cells,and then were cultivated and serial subcultivated in vitro.P3 cultures were analyzed by the flow cytometry to determine the surface antigens,and expressive intensity of melatonin receptors(MT1,MT2)was detected by RT-PCR.Results:Mononuclear cells were cultivated and subcultivated to P3 culture in vitro,which were analyzed by the flow cytometry,and demonstrated that the expanded mononuclear cells expressed mesenchymal cell markers.There was no statistical difference of melatonin receptors expression in BMSCs between AIS group and control(P > 0.05).Conclusion:The expression of melatonin receptors in BMSCs were comparable between AIS patients and controls,which indicated that melatonin receptors would not involved in the pathogenesis of AIS in the stem cell level.