针刺血清对体外培养神经细胞内钙离子浓度的影响

目的:通过观察针刺大鼠经穴后收集的血清对体外培养的神经细胞内Ca2 浓度的影响,为探讨针灸作用的机制中是否存在着体液因素提供直接的证据.方法:取新生大鼠大脑皮层细胞进行原代培养.7～10 d后应用荧光分子探针Fluo-3AM染色,激光共焦扫描显微镜动态扫描观察加入正常大鼠血清和电针刺激百会、足三里、曲池、三阴交2周后的血清对神经细胞内Ca2 浓度的影响.结果:加入正常血清后,细胞内的Ca2 浓度会有一定的升高,经过一段时间后升高的Ca2 浓度会趋向平稳.而再加入针刺血清后,在一定程度上可以降低神经细胞Ca2 的浓度.结论:针刺腧穴后,除了神经的调节机制外,针刺血清中存在某种活性物质,可以明显地使神经细胞内Ca2 浓度降低,为针灸体液因素的存在提供了直接的证据.

Effect of acupuncture serum on intracellular Ca2+ concentration in cultured neurons

OBJECTIVE: To study the humoral factor of acupuncture in improving functional activities of the organism. METHODS: Neurons of the cerebral cortex from the new born rats were taken to be cultured in a strictly controlled medium. Seven to ten days later, fluorescein-molecular probe Fluo-3AM was used for staining intracellular Ca(2+). Ca(2+) concentration and its changes after application of acupuncture serum were measured by using a laser confocal microscope. Normal serum and acupuncture serum were derived from the normal control rats and the rats undergoing electro-acupuncture of "Baihui (GV 20)", "Zusanli (ST 36)", "Quchi (LI 11)" and "Sanyinjiao (SP 6)" for 2 weeks. RESULTS: After application of normal serum, the intracellular Ca(2+) concentration increased strikingly immediately at the beginning, and gradually got stable afterwards. When acupuncture serum was added to the culture fluid, Ca(2+) concentration lowered significantly. CONCLUSION: These results suggest that bioactive substances in serum collected from rats undergoing acupuncture can affect the intracellular Ca(2+) concentration of the cerebral neurons, and the humoral factor may participate in the effect of acupuncture in regulating functional activities of the organism.