电针干预对脑梗死大鼠脑组织Wnt信号通路的影响  

Effect of electroacupuncture on neurological function and Wnt signaling pathway in ischemic brain tissue of cerebral infarction rats

作　　者：张晶晶 杜元灏 李晶[2] 杨丽红 陈林玲 查庆平 ZHANG Jing-jing;DU Yuan-hao;LI Jing;YANG Li-hong;CHEN Lin-ling;ZHA Qing-ping(Department of Acupuncture and Moxibustion,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300112,China;Institute of Acupuncture and Moxibustion,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300112,China)

机构地区：[1]天津中医药大学第一附属医院针灸科,天津300112 [2]天津中医药大学第一附属医院针灸研究所,天津300112

出　　处：《针刺研究》2020年第3期202-208,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81473765);天津市教委科研计划项目(自然科学,No.2018KJ025)。

摘　　要：目的:观察电针干预对脑梗死(CI)大鼠脑组织Wnt信号蛋白中7a(Wnt7a)、淋巴增强因子1(LEF1)、糖原合成酶激酶3β(GSK-3β)、Wnt信号通路内源性抑制剂Dickkopf-1(DKK1)mRNA及蛋白表达的影响,探讨电针治疗CI的作用机制。方法:将280只健康雄性Wistar大鼠随机分为电针组、模型组、假手术组和空白组,前3组每组90只,并按照术后1 h、3 h、6 h、9 h、12 h、24 h、3 d、7 d、12 d分为9个时相组,每组10只,空白组10只。采用改良的Longa法建立永久性大脑中动脉闭塞(MCAO)大鼠模型。电针组予电针刺激"水沟"穴,疏密波,2 Hz/15 Hz,2 mA,留针20 min,1、3、6、9、12、24 h时相组大鼠电针治疗1次,其余时相每日电针1次。采用神经损伤严重程度(NSS)评分于术前、术后及取材前对各组大鼠进行评价。采用实时荧光定量PCR法及Western blot法检测各组大鼠右侧缺血区脑组织Wnt7a、LEF1、GSK-3β及DKK1 mRNA和蛋白表达水平。结果:MCAO后,模型组及电针组大鼠NSS评分随缺血时间延长呈逐渐下降趋势;取材前,与假手术组比较,模型组大鼠各时相NSS评分明显升高(P<0.01);与模型组比较,电针组大鼠NSS评分在3、7、12 d时明显降低(P<0.05,P<0.01)。与假手术组比较,模型组大鼠缺血区脑组织Wnt7a和LEF1 mRNA表达水平在3 h^12 d时均明显升高(P<0.01,P<0.05),GSK-3β和DKK1 mRNA表达水平分别在9~24 h和24 h^3 d时明显降低(P<0.05,P<0.01);模型组Wnt7a和LEF1蛋白表达水平在6 h^12 d均明显升高(P<0.05,P<0.01),GSK-3β和DKK1蛋白表达水平分别在24 h^3 d和3 d时明显降低(P<0.01,P<0.05)。与模型组比较,电针组大鼠缺血区脑组织Wnt7a和LEF1 mRNA表达水平分别在12 h^3 d和24 h^12 d时明显升高(P<0.01,P<0.05),GSK-3β和DKK1 mRNA表达水平分别在9 h、3~12 d和12 h^3 d时明显降低(P<0.01,P<0.05);电针组Wnt7a和LEF1蛋白表达水平分别在24 h^12 d和3~12 d明显升高(P<0.05,P<0.01),GSK-3β和DKK1蛋白表达水平分别在12 h、7~12 d和24 h^12 dObjective To explore the mechanism of electroacupuncture(EA) underlying improvement of cerebral infarction(CI) by investigating its influence on expression of cerebral Wnt7 a, lymphoid enhancer factor-1(LEF1), glycogen synthase kinase 3β(GSK-3β) and Dickkopf-1(DKK1) mRNA and proteins in CI rats. Methods A total of 280 male Wistar rats were randomly divided into blank control(n=10), sham-operation, model and EA groups,and 90 rats of the last 3 groups were further divided into 1, 3, 6, 9, 12 and 24 h, and 3, 7 and 12 d subgroups with 10 rats in each subgroup. The CI model was established by occlusion of the middle cerebral artery(MCAO). The sham-operation group received the same surgical operation but without thread embolus insertion. EA(2 Hz/15 Hz, 2 mA) was applied to "Shuigou"(GV26) for 20 min, once a day for 1, 3, 7 and 12 d, respectively. The neurological deficit was evaluated by using Neurological Severity Scores(NSS). The expression levels of Wnt7 a,LEF1, GSK-3β and DKK1 mRNAs and proteins in the right ischemic brain tissues were detected by Quantative real-time PCR and Western blot, respectively. Results After MCAO, the NSS score was significantly increased in the model and EA groups relevant to the blank control and sham-operation groups(P<0.01) and gradually decreased with the prolongation of ischemia time. After EA, the NSS scores were notably decreased on day 3, 7 and 12 in the EA group compared with the model group(P<0.05, P<0.01). After modeling, the expression levels of Wnt7 a and LEF1 mRNAs from 3 h to 12 d, Wnt7 a and LEF1 proteins from 6 h to 12 d were considerably increased(P<0.01, P<0.05), while those of GSK-3β mRNA at 9, 12 and 24 h, GSK-3β protein at 24 h and 3 d, and DKK1 mRNA at 24 h and 3 d and DKK1 protein at 3 d were obviously decreased in the model group relevant to the sham-operation group(P<0.05, P<0.01). After the intervention, the expression levels of Wnt7 a mRNA at 12 h to 3 d, Wnt7α protein from 24 h to 12 d, LEF1 mRNA from 24 h to 12 d, and LEF1 protein from 3 d to 12 d were

关 键 词：脑梗死 电针 水沟 神经功能 WNT信号通路 

分 类 号：R245.97[医药卫生—针灸推拿学]