中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2010年
2期
124-127
,共4页
王斌%李海波%邱勇%邱旭升%朱锋%孙光权%王渭君
王斌%李海波%邱勇%邱旭升%硃鋒%孫光權%王渭君
왕빈%리해파%구용%구욱승%주봉%손광권%왕위군
褪黑激素%信号转导%间充质干细胞%骨髓%青少年特发性脊柱侧凸
褪黑激素%信號轉導%間充質榦細胞%骨髓%青少年特髮性脊柱側凸
퇴흑격소%신호전도%간충질간세포%골수%청소년특발성척주측철
Melatonin%Signal transduction%Mesenchymal stem cells%Bone marrow%Adolescent idiopathic scoliosis
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.
目的 研究青少年特髮性脊柱側凸(Ms)患者骨髓間充質榦細胞(BMSCs)褪黑素信號通路是否存在異常.方法 24例12～18歲誌願者,AIS患者15例,正常對照組9例.分彆從患者髂前上棘處穿刺抽取10 ml骨髓,肝素抗凝.採用密度梯度離心法分離BMSCs,體外培養併傳至P3代,進行錶型鑒定.取P3代BMSCs,先採用福斯可林刺激BMSCs內環燐痠腺苷(cAMP)升高,然後用不同濃度褪黑素刺激細胞,檢測細胞內cAMP的水平.結果 分離的單箇覈細胞經培養至P3代,使用流式細胞儀進行細胞錶型鑒定,體外培養細胞的錶型與BMSCs的錶麵標誌相符.AIS組和正常對照組BMSCs的cAMP的基礎水平都很低,在給予福斯可林刺激以後,cAMP的水平明顯升高,此後再給予生理劑量的褪黑素刺激後,兩組cAMP水平的抑製程度差異無統計學意義(P>0.05);給予藥理劑量的褪黑素刺激後,兩組cAMP水平的抑製程度差異同樣無統計學意義(P>0.05).結論 MS患者BMSCs褪黑素信號通路可能不存在異常.
목적 연구청소년특발성척주측철(Ms)환자골수간충질간세포(BMSCs)퇴흑소신호통로시부존재이상.방법 24례12～18세지원자,AIS환자15례,정상대조조9례.분별종환자가전상극처천자추취10 ml골수,간소항응.채용밀도제도리심법분리BMSCs,체외배양병전지P3대,진행표형감정.취P3대BMSCs,선채용복사가림자격BMSCs내배린산선감(cAMP)승고,연후용불동농도퇴흑소자격세포,검측세포내cAMP적수평.결과 분리적단개핵세포경배양지P3대,사용류식세포의진행세포표형감정,체외배양세포적표형여BMSCs적표면표지상부.AIS조화정상대조조BMSCs적cAMP적기출수평도흔저,재급여복사가림자격이후,cAMP적수평명현승고,차후재급여생리제량적퇴흑소자격후,량조cAMP수평적억제정도차이무통계학의의(P>0.05);급여약리제량적퇴흑소자격후,량조cAMP수평적억제정도차이동양무통계학의의(P>0.05).결론 MS환자BMSCs퇴흑소신호통로가능불존재이상.
Objective 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.
