电针“内关”和“足三里”对脑梗死大鼠神经损伤修复及Orexin-A、炎性因子表达的影响

Effect of electroacupuncture at“Neiguan”and“Zusanli”on nerve-injury repair and the expressions of serum Orexin-A and inflammatory factors in rats with cerebral infarction

目的:观察电针“内关”和“足三里”对脑梗死大鼠神经损伤修复及食欲素A(Orexin-A)、血清炎性因子表达的影响。方法:54只成年雄性SPF级大鼠随机分为假手术组、模型组和电针组,电针组和模型组制备大脑中动脉闭塞(MCAO)模型,24 h后电针组大鼠取双侧“内关”和“足三里”电针治疗14 d。假手术组仅做颈部皮肤切开缝合处理。各组做神经功能缺损评分、称重,取材后行TTC、HE和尼氏染色,蛋白印记和荧光定量PCR检测Orexin-A表达。采用酶联免疫吸附试验(ELISA)法检测血清Orexin-A、血清炎性因子(TNF-α、IL-6、IL-1β)等。结果:与假手术组相比,模型组神经功能缺损评分和脑梗死体积升高,体重降低(P<0.05),模型组神经元数量、血清Orexin-A表达和脑组织Orexin-A蛋白及mRNA表达量明显降低,血清炎性因子水平明显升高(P<0.01);与模型组相比,电针组神经功能缺损评分和脑梗死体积降低,体重升高(P<0.05),电针组神经元数量、血清Orexin-A表达和脑组织Orexin-A蛋白及mRNA表达量明显升高,血清炎症因子水平明显降低(P<0.01)。模型组大鼠脑组织完整神经细胞数量减少,细胞胞核分离,电针组神经细胞形态相对完整,趋于正常。结论:电针疗法可以改善脑梗死大鼠的神经功能缺损情况,减小脑梗死体积,改善体重情况,其机制可能与调节Orexin-A水平,抑制血清炎性因子产生,促进神经损伤修复有关。

Objective:To observe the effect of electroacupuncture(EA)at“Neiguan”and“Zusanli”on nerve-injury repeir and the expressions of Orexin-A and serum inflammatory factors in rats with cerebral infarction.Methods:Fifty-four adult male SPF rats were randomly divided into sham operation group,model group and EA group.Middle cerebral artery occlusion(MCAO)model was established in the EA group and model group.Twenty-four hours later,EA was applied to the rats in EA group at bilateral“Neiguan”and“Zusanli”for 14 days.The sham operation group was only treated with neck skin incision and suture.After sampling,TTC,HE and Nissl staining were performed.Western blotting and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)were used to detect Orexin-A.ELISA was used to detect serum Orexin-A and serum inflammatory factors(TNF-α,IL-6,IL-1β).Results:Compared with the sham operation group,the neurological deficit score and cerebral infarction volume increased,and the body weight decreased in the model group(P<0.05);the number of neurons,serum Orexin-A expression and the expressions of Orexin-A protein and mRNA in brain tissue significantly decreased,and the serum inflammatory factors significantly increased in the model group(P<0.01).Compared with the model group,the neurological deficit score and cerebral infarction volume decreased,and the body weight increased in the EA group(P<0.05);the number of neurons,serum Orexin-A level and the expression of Orexin protein and mRNA expression in brain tissue significantly increased,and the serum inflammatory factors significantly decreased in the EA group(P<0.01).In the model group,the number of intact nerve cells in brain tissue was reduced,and the cell nuclei were separated,while the morphology of nerve cells in the EA group was relatively complete and tended to be normal.Conclusion:EA therapy can improve the neurological deficit of rats with cerebral infarction,reduce the cerebral infarction volume and improve the body weight.Its mechanism may be related to the regulation of Orexin-A level,inhibition of serum inflammatory factors and promotion of nerve repair.