电针“阳陵泉”“曲池”对脑卒中肢体痉挛大鼠大脑皮质γ-氨基丁酸能系统的影响

Effects of Electroacupuncture on“Yanglingquan”and“Quchu”for GABAergic System in Cerebral Cortex of Rats with Post-stroke Spasticity

目的观察电针“阳陵泉”“曲池”对脑卒中肢体痉挛(PSS)大鼠大脑皮质γ-氨基丁酸(GABA)能系统相关因子的影响,探讨其治疗PSS的可能机制。方法采用右侧大脑中动脉线栓+内囊注射N-甲基-D-天冬氨酸(NMDA)受体法制备PSS模型。将大鼠随机分为假手术组、模型组和电针组,每组10只。电针组选取患侧“阳陵泉”“曲池”电针干预,30 min/次,1次/d,连续7 d。假手术组和模型组仅抓取固定、不进行干预。对大鼠进行神经功能缺损评分和肌张力评分,电生理描记法测定大鼠肌张力,Western blot和RT-qPCR检测缺血侧大脑皮质GABA能系统合成因子谷氨酸脱羧酶65(GAD65)、重摄因子GABA转运蛋白-3(GAT-3)、传导因子钠-钾-氯协同转运蛋白1(NKCC1)蛋白及mRNA表达。结果与假手术组比较,模型组大鼠神经功能缺损评分、肌张力评分明显升高(P<0.01),肌张力明显增加(P<0.01),缺血侧大脑皮质GAD65、GAT-3蛋白及mRNA表达明显降低(P<0.01),NKCC1蛋白及mRNA表达明显升高(P<0.01);与模型组比较,电针组大鼠神经功能缺损评分、肌张力评分明显降低(P<0.05),肌张力明显减小(P<0.01),缺血侧大脑皮质GAD65、GAT-3蛋白及mRNA表达明显升高(P<0.01,P<0.05),NKCC1蛋白及mRNA表达明显降低(P<0.05,P<0.01)。结论电针“阳陵泉”“曲池”能改善PSS大鼠神经功能缺损症状,降低肌张力,缓解卒中后肢体痉挛。其机制可能是通过促进模型大鼠大脑皮质GABA合成酶GAD65、转运体GAT-3表达,抑制转运蛋白NKCC1表达,从而维持GABA能系统稳态。

Objective To observe the effects of electroacupuncture on“Yanglingquan”and“Quchi”for the GABAergic system pathway in the cerebral cortex of rats with post-stroke spasticity(PSS);To explore the possible mechanism of electroacupuncture in the treatment of PSS.Methods The rat model of PSS was prepared by using right middle cerebral artery thread thrombus method+internal capsule injection of NMDA receptor.The rats were randomly divided into sham-operation group,model group and electroacupuncture group,with 10 rats in each group.The electroacupuncture group chose“Yanglingquan”and“Quchi”on the affected side for treatment,and the treatment was carried out once a day for 30 min each time for 7 consecutive days.The sham operation group and the model group were only grasped and fixed without other interventions.Neurological deficit and muscle tone scores were performed on rats.Muscle tone in rats was detected using electrophysiological recording method.Western blot and RT-qPCR were used to detect GABAergic system composition factor glutamate decarboxylase 65(GAD65),reabsorption factor GABA transporter-3(GAT-3),conduction factor sodium potassium chloride cotransporter-1(NKCC1)protein and mRNA expression in the ischemic cerebral cortex.Results Compared with the sham-operation group,the neurological deficit score and muscle tone score of the model group rats significantly increased(P<0.01),while muscle tone was significantly increased(P<0.01),the expressions of GAD65,GAT-3 protein and mRNA in the ischemic cerebral cortex were significantly decreased(P<0.01),the expression of NKCC1 protein and mRNA significantly increased(P<0.01).Compared with the model group,the neurological deficit score and muscle tension score of the electroacupuncture group were significantly decreased(P<0.05),while muscle tone was significantly decreased(P<0.01),the expressions of GAD65,GAT-3 protein and mRNA in the ischemic cerebral cortex significantly increased(P<0.01,P<0.05),the expression of NKCC1 protein and mRNA significantly decreased(P<0.05,P<0.01).Conclusion Electroacupuncture on“Yanglingquan”and“Quchi”can effectively improve the symptoms of neurological deficits in PSS rats,decrease muscle tone,and alleviate spasticity after stroke.The mechanism may be to promote the expression of GABA synthase GAD65 and transporter GAT-3 in the cerebral cortex of model rats,inhibit the expression of transport protein NKCC1,and maintain the homeostasis of the GABAergic system.