电针血清对体外培养小胶质细胞活性的影响

Effect of electroacupuncture on microglia activation in vitro

目的:观察电针血清对体外培养的小胶质细胞活性和一氧化氮释放量的影响,探讨电针治疗帕金森病的机制。方法:采用高频电针针刺正常大鼠,2周后制备电针血清;体外培养的小胶质细胞分别加入正常血清(正常组)、正常血清加入LPS(对照组)和电针血清加入LPS(电针组)培养;通过免疫组化、MTT和Griess法分别观测8～48h不同时间小胶质细胞形态、活性及NO释放量。结果:1μg/ml LPS可激活小胶质细胞,表现为细胞体积增大,OX-42表达上调,电针组12h小胶质细胞的活性低于对照组,并且12～48 h NO释放量均显著低于对照组,增加电针血清浓度可显著减少NO的释放量。结论:电针血清可能通过抑制激活的小胶质细胞释放NO,从而保护多巴胺神经元。

Objective： To explore the mechanism of electroacupuncture treatment on Parkinson disease, and investigate the effect of electroacupuncture on microglia activation in vitro and NO release. Methods： 100 Hz high frequency electroacupuncture was used to stimulate Baihui, Fengfu acupuncture points in rats. After 2 weeks, the serum was observed. The microglia was divided into three groups： normal group, added with normal rat serum; control group, added with normal rat serum and microglia activator LPS （1 μg/ml） . and experimental group, added with the serum of the electroacupuncture treated rats and LPS （1 μg/ml）. 8 h, 12 h, 24 h and 48 h after treatment, the morphology, viability and NO release of microglia were respectively detected by immunostaining, MTT and Griess methods. Results： 1 μg/ml LPS could activate microglia, indicated by the increased soma, up-regulation of OX-42 and increased NO release in comparison with the control group. No significant increase, however, was found in the number of cells. 12 h after treatment, the viability of microglia in the experimental group was less than that in the control group. The NO release in the experimental group was significantly less than that in the control group at 12 h, 24 h and 48 h （P〈0. 05）, and this effect was dose dependent. Conclusion： Electroacupuncture may inhibit NO release from activated microglia to protect do-paminergic neuron.