推拿法样刺激对人脐静脉内皮细胞内钙离子浓度的影响  

Effects of stimulus like rolling manipulation on the intracellular calcium concentration in human umbilical vein endothelial cells

作　　者：马惠昇[1] 张宏[2] 严隽陶[2] 赵谦[2] 牛坤[2] 

机构地区：[1]宁夏医科大学中医学院,宁夏回族自治区银川市750004 [2]上海中医药大学附属岳阳中西医结合医院,上海市200437

出　　处：《中国组织工程研究与临床康复》2010年第37期6887-6890,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然基金资助项目(30300462)"推拿法操作动力学参数优化及行气活血效应机制研究";上海市教育委员会科研创新项目(09YZ138)"不同人群推拿法操作优化及舒筋活络效应机理研究";上海市针灸推拿学重点学科建设项目(T0302-A24)"推拿法舒筋活络效应的操作规范化及作用机理研究~~

摘　　要：背景:迄今对推推拿法作用机制的研究多着眼于软组织、骨关节、神经、血管等宏观层面,还需要从微观细胞学水平对手法力学刺激钙离子通道引发的效应机制进行研究。目的:观察推拿法样刺激对人脐静脉内皮细胞内钙离子浓度的影响。方法:将正常培养和缺氧培养的人脐静脉内皮细胞均分为对照组、推拿组、维拉帕米组。对照组细胞不做任何处理,推拿组细胞用Fluo-3/AM标记后给予手法样压力刺激,压力大小为0.4kg,频率120次/min,时间1min;维拉帕米组细胞用Fluo-3/AM标记后,先加入终浓度为10-5mol/L的维拉帕米,然后给与同样的法样刺激。各组细胞在处理前后分别采用共聚焦激光扫描显微镜观察并测定细胞荧光强度进行比较。结果与结论:手法样刺激和缺氧环境均可引起人脐静脉内皮细胞内游离钙离子浓度升高,但这两种促进人脐静脉内皮细胞内游离钙离子浓度升高的作用均能够被钙离子拮抗剂维拉帕米所抑制。由此推测,推拿法的作用机制是通过机械刺激激活钙离子通道,引发细胞钙离子内流,促使细胞内游离钙离子浓度升高,进而启动并产生后续的生物学治疗效应。BACKGROUND：More researches of rolling manipulation in traditional Chinese Medicine focused on the macroscopic mechanism of action on soft tissues,joints,nerves,and blood vessels.But reports about its effect on cells such as the calcium ionchannels are few.OBJECTIVE：To observe the effect of rolling manipulation on the intracellular calcium concentration of human umbilical vein endothelial cells（HUVEC）.METHODS：The normal and hypoxic HUVEC were divided into the control,massage therapy,massage ＋ verapamil group.The control group had no interference.Cells in the massage therapy group were marked with Fluo-3/AM,and then were stimulated with 0.4 kg in pressure,120 times per minutes in frequency,and maintained for 1 minute.The massage ＋ verapamil group cells were marked with Fluo-3/AM,and the final concentration of 10-5 mol/L of the verapamil were added in their culture fluid.They were stimulated as the massage therapy group cells.Each group cells were observed and measured the fluorescence intensity by the confocal laser scanning microscope before and after treatment.RESULTS AND CONCLUSION：Both stimulus like rolling manipulation and hypoxic environment could increase the intracellular calcium ion concentration,but they can be prevented by verapamil.Accordingly,it is presume that the mechanism of action of rolling manipulation is activating Ca2＋ channels,causing Ca2＋ internal flow and then increasing the intracellular Ca2＋,which primings its therapeutic effects.

关 键 词：推拿法 钙离子 人脐静脉内皮细胞 信号转导 组织工程 

分 类 号：R318[医药卫生—生物医学工程]