耳针血清对大鼠脑微血管内皮细胞拟糖尿病损伤的保护作用

Efficacy of auricular acupuncture serum on morphological change of brain microvessel endothelial cells induced by diabetic incubation in vitro

目的从胆碱能抗炎通路探讨耳针血清对大鼠脑微血管内皮细胞拟糖尿病损伤的保护作用。方法耳针穴位刺激2型糖尿病大鼠模型,并取血清,体外培养大鼠脑微血管内皮细胞株,细胞分别培养于不同血清条件的培养液中。采用MTS/PMS比色分析法测定各组细胞的活性,流式细胞仪观察内皮细胞的凋亡情况,透射电镜观察各组细胞的超微结构变化。结果耳针血清能明显增加细胞活性、显著降低糖尿病引起的细胞凋亡(P均<0.01);改善内皮细胞的超微结构受损情况。但迷走神经阻断后针刺大鼠,其血清培养的细胞活性明显低于针刺组(P<0.01);细胞凋亡率显著高于针刺组(P<0.05)。结论耳针血清对血管内皮细胞具有显著的形态学保护作用,其作用机制可能与兴奋迷走神经有关。

Objective To investigate the effects of auricular acupuncture serum on the injury of brain microvessel endothelial cells induced by diabetic incubation in vitro in the light of cholinergic anti-inflammatory pathway. Methods Type-2-model diabetes rats were treated with electro auricular acupuncture and rat serum samples were collected. Brain microvessel endothelial cells were cultured with differ ent incubations in vitro and treated with different conditions of culture medium. Cell viabilities in each group were assayed with MTS/PMS. Apoptosis of ceils in each group were detected using flow cytometry. The ultrastructural alterations of microvessel endothelial cells were ob served with transmission electron microscope. Results Auricular acupuncture serum significantly （P 〈 0. 01 ） increased cell viabilities of vascular endothelial ceils and remarkably （P 〈0. 01 ） inhibited the apoptosis of vascular endothelial ceils induced by diabetic incubation in vitro, and improved the damage of ultrastructures of microvessel endothelial cells. However, the cell viabilities in the group of blocked vagus nerve were significantly lower than the acupuncture group （P 〈0. 01 ） and the apoptosis rate was significantly higher than those in the acu- puncture group （P 〈 0. 05）. Conclusions Auricular acupuncture serum can significantly protect against vascular endothelial cells injury in duced by diabetic incubation, and the effect may be mediated by stimulation of vagas nerve.