摘要目的 研究青少年特发性脊柱侧凸(Ms)患者骨髓间充质干细胞(BMSCs)褪黑素信号通路是否存在异常.方法 24例12～18岁志愿者,AIS患者15例,正常对照组9例.分别从患者髂前上棘处穿刺抽取10 ml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.取P3代BMSCs,先采用福斯可林刺激BMSCs内环磷酸腺苷(cAMP)升高,然后用不同浓度褪黑素刺激细胞,检测细胞内cAMP的水平.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组和正常对照组BMSCs的cAMP的基础水平都很低,在给予福斯可林刺激以后,cAMP的水平明显升高,此后再给予生理剂量的褪黑素刺激后,两组cAMP水平的抑制程度差异无统计学意义(P>0.05);给予药理剂量的褪黑素刺激后,两组cAMP水平的抑制程度差异同样无统计学意义(P>0.05).结论 MS患者BMSCs褪黑素信号通路可能不存在异常.

abstractsObjective To investigate the melatonin signaling transduction pathway in BMSCs from adolescent idiopathic scoiiosis patients. Methods Twenty-four volunteers aged 12-18 years were divided into two groups; AIS group was 15 and control group was 9. 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. P3 BMSCs were used to detect the melatonin signaling transduction pathway. The cellular cAMP was elevated using forskolin, and then the BMSCs were treated with melatonin to inhibit the cellular cAMP levels. 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. The basal cAMP levels of the two groups were very low, after the stimulation of forskolin, cellular cAMP levels increased rapidly in all the patients, but after the stimulation of melatonin at physiological dose or even at pharmacological dose, there was no statistical difference of the inhibition of cAMP between AIS group and control (P > 0. 05). Conclusion Melatonin signaling transduction pathway may be normal in BMSCs from AIS patients.