电针调节Th17/Treg细胞平衡对脑梗死大鼠神经炎症的影响

Regulatory effects of electroacupuncture on neuroinflammation in rats with cerebral infarction by regulating the balance of Th17/Treg cells

目的 探究电针对脑梗死大鼠的保护作用及其潜在的抗神经炎症机制。方法 建立大鼠中动脉闭塞/再灌注(MCAO/R)模型,随机分为模型组、电针非穴位组、电针治疗(水沟穴)组,另设假手术组,每组24只大鼠。末次电针治疗后,使用神经行为评估和TTC染色来检查脑损伤情况;流式细胞术检测外周血Th17/Treg细胞水平;HE染色和Nissl染色评估电针对神经元损伤的影响;脑含水量和水通道蛋白4(AQP4)检测用于评估脑水肿;免疫组织化学评估小胶质细胞和星形胶质细胞活化;Western blot评估炎症反应和Th17细胞因子(IL-17)、Treg细胞因子(IL-10)水平。结果 与假手术组比较,模型组大鼠神经学评分和脑梗死体积增高,神经元损伤和脑水肿程度增加,小胶质细胞和星形胶质细胞活化增加,炎症反应、Th17细胞比例和IL-17水平上调,Treg细胞比例和IL-10水平下调(P<0.05)。与模型组比较,电针非穴位组大鼠以上指标变化无统计学意义(P>0.05);与电针非穴位组比较,电针治疗组大鼠可明显改善MCAO/R模型大鼠的以上症状(P<0.05)。结论 电针治疗可调节脑梗死大鼠Th17/Treg细胞平衡,抑制小胶质细胞活化和神经炎症。

To explore the protective effect of electroacupuncture(EA) on rats with cerebral infarction, and to explore its potential anti-neuroinflammatory mechanism, a model of middle artery occlusion/reperfusion(MCAO/R)was established in rat, which were then randomly divided into model group, non-acupoint site electroacupuncture group, electroacupuncture treatment(Shuigou point) group, while another sham operation group was set, with 24animals in each group. After last EA treatment, the neurobehavioral assessment and TTC staining were used to examine brain injury, and flow cytometry was used to detect the level of Th17/Treg cells in peripheral blood. HE staining and Nissl staining were performed to evaluate the effect of EA on neuronal damage;brain water content and aquaporin 4(AQP4) were measured to assess cerebral edema;immunohistochemistry was performed to assess microglia and astrocyte activation;Western blot was performed to assess the inflammatory response and Th17/Treg balance. Compared with the sham operation group, the neurological score and cerebral infarction volume of the rats in the model group increased, the degrees of neuronal damage and brain edema were increased, the activation of microglia and astrocytes were increased, the inflammatory response, Th17 cell proportion and IL-17 level were increased, Treg cell proportion and IL-10 level was up-regulated, and the level of Treg cells was down-regulated(P<0.05). Compared with the model group, the aboveindexes in the EA non-acupoint group were notsignificantly different(P>0.05). While compared with the EA non-acupoint group, the EA treatment group demonstrated great alleviation in the above mentionedsymptoms of the MCAO/R model rats(P<0.05). Taken together, electroacupuncture can regulate the balance of Th17/Treg cells, inhibit microglia activation and neuroinflammation in rats with cerebral infarction.