姚小强;李兴兰;杜小正;王金海;袁博;张婷卓;张枫帆;乔翔;王一心;
YAO Xiao-qiang;LI Xing-lan;DU Xiao-zheng;WANG Jin-hai;YUAN Bo;ZHANG Ting-zhuo;ZHANG Feng-fan;QIAO Xiang;WANG Yi-xin;Acupuncture Clinical Center,Affiliated Hospital of Gansu University of Chinese Medicine;College of Acupuncture-moxibustion and Tuina,Gansu University of Chinese Medicine;Department of Traditional Chinese Medicine,the Second Hospital of Lanzhou University;

• 1:甘肃中医药大学附属医院针灸中心
• 2:甘肃中医药大学针灸推拿学院
• 3:兰州大学第二医院中医科

摘要(Abstract)：

目的:观察头穴透刺对局灶性脑缺血大鼠纹状体区正五聚环蛋白3(PTX3)、白细胞介素(IL)-1β、闭锁小环蛋白-1(ZO-1)mRNA及咬合蛋白(Occludin)mRNA表达的影响,探讨头穴透刺维护血脑屏障完整性的机制。方法:SD大鼠随机分为正常组、模型组、IL-1受体拮抗剂(IL-1Ra)组、IL-1Ra+头针组,每组12只。采用大脑中动脉线栓法建立大脑中动脉栓塞(MCAO)大鼠模型。IL-1Ra组在造模成功后1 h予腹腔注射IL-1Ra(0. 05 mg/kg),1次/d,共6 d。IL-1Ra+头针组在造模成功后1 h予腹腔注射IL-1Ra(0. 05 mg/kg),并予双侧顶颞前斜线快速捻转透刺治疗,1次/d,共6 d。干预前后进行神经功能缺损评分;免疫组织化学法测定纹状体区PTX3、IL-1β的表达;荧光定量PCR法检测纹状体区ZO-1、Occludin mRNA的表达;伊文思蓝(EB)示踪法监测血脑屏障损伤程度。结果:与正常组比较,模型组大鼠神经功能缺损评分、纹状体区IL-1β和PTX3的表达、脑内EB含量明显增高(P<0. 05),纹状体区ZO-1、Occludin mRNA的表达明显降低(P<0. 05)。与模型组比较,两个干预组大鼠神经功能缺损评分、脑内EB含量明显降低(P<0. 05),纹状体区ZO-1、Occludin mRNA的表达明显升高(P<0. 05);IL-1Ra组大鼠纹状体区PTX3表达明显降低(P<0. 05);IL-1Ra+头针组大鼠纹状体区IL-1β表达明显降低(P<0. 05),PTX3的表达明显升高(P<0. 05)。与IL-1Ra组比较,IL-1Ra+头针组神经功能缺损评分、脑内EB含量均明显降低(P<0. 05),大鼠纹状体区IL-1β的表达明显降低(P<0. 05),PTX3的表达明显升高(P<0. 05),ZO-1、Occludin mRNA的表达明显升高(P<0. 05)。结论:上调缺血后纹状体区PTX3的表达,促进ZO-1、Occludin mRNA的表达,减轻血脑屏障损伤程度,可能是头穴透刺改善缺血性脑损伤的作用机制之一。
Objective To observe the influence of scalp-acupuncture on the expression of pentraxin 3(PTX3),Interleukin(IL)-1β,zonula occludens-1(ZO-1)mRNA and Occludin mRNA in striatum in acute ischemic cerebrovascular disease(AICD)rats,so as to investigate its mechanisms underlying improvement of AICD.MethodsForty-eight male SD rats were randomly allocated to control,model,IL-1 Ra and IL-1 Ra+scalp-acupuncture groups(n=12 rats in each group). The AICD model was established by occlusion of the middle cerebral artery(MCAO). Rats of the IL-1 Ra group and IL-1 Ra+scalp-acupuncture group received intraperitoneal injection of IL-1 Ra(0. 05 mg·kg^(-1)·d(-1)·d^(-1)),once daily for 6 days. Scalp acupuncture stimulation was applied to bilateral"Dingnieqianxiexian"(MS6)once daily for 6 days for rats in IL-1 Ra+scalp-acupuncture group. Before and after intervention,the neurologic deficit score(NDS)was evaluated according to Longa's method. The expression of striatum PTX3 and IL-1β was detected by immunohistochemistry,and ZO-1 mRNA and Occludin mRNA in the striatum tissue were detected by fluorescence quantitative real-time PCR.The Evans Blue(EB)tracer method was used to monitor the degree of blood-brain barrier(BBB)damage.ResultsFollowing modeling,the NDS,EB content and the expression of PTX3 and IL-1β in the striatum tissue were significantly increased,and the ZO-1 mRNA and Occludin mRNA expression was considerably decreased in the model group compared with the control group(P < 0. 05). After the interventions and compared with the model group,the NDS,EB content in both IL-1 Ra and IL-1 Ra+scalp acupuncture groups,and PTX3 in the IL-1 Ra group were significantly downregulated(P<0. 05),while the striatum ZO-1 mRNA and Occludin mRNA expression in both IL-1 Ra and IL-1 Ra+scalp acupuncture groups,and PTX3 in the IL-1 Ra+scalp acupuncture group were obviously up-regulated(P<0. 05),and the expression of IL-1β was obviously down-regulated in the IL-1 Ra+scalp acupuncture group(P<0. 05)rather than in the IL-1 Ra group(P>0. 05). The effects of scalp acupuncture combined with IL-1 Ra were obviously superior to that of IL-1 Ra in down-regulating NDS,EB content and IL-1β expression level,and in up-regulating PTX3,ZO-1 mRNA and Occludin mRNA expression levels(P<0. 05).ConclusionScalp acupuncture can improve neurological function and reduce the degree of BBB injury in AICD rats,which may be associated with its function in up-regulating the expression of PTX3 and in promoting the expression of ZO-1 mRNA and Occludin mRNA.

关键词(KeyWords)： 头穴透刺;急性缺血性中风;正五聚环蛋白3;白细胞介素-1β;闭锁小环蛋白-1;咬合蛋白;白细胞介素1受体拮抗剂
Scalp acupuncture;Acute ischemic stroke;Pentraxin 3;Interleukin-1β;Zonula occludens-1;Occludin;Interleukin-1Ra

Abstract:

Keywords:

基金项目(Foundation): 甘肃省科技厅创新基地和人才计划项目(No.18JR3RA196)

作者(Authors): 姚小强;李兴兰;杜小正;王金海;袁博;张婷卓;张枫帆;乔翔;王一心;
YAO Xiao-qiang;LI Xing-lan;DU Xiao-zheng;WANG Jin-hai;YUAN Bo;ZHANG Ting-zhuo;ZHANG Feng-fan;QIAO Xiang;WANG Yi-xin;Acupuncture Clinical Center,Affiliated Hospital of Gansu University of Chinese Medicine;College of Acupuncture-moxibustion and Tuina,Gansu University of Chinese Medicine;Department of Traditional Chinese Medicine,the Second Hospital of Lanzhou University;

DOI: 10.13702/j.1000-0607.180899

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