王金海;杜小正;张婷卓;李兴兰;赵敏;鲍英存;鄢琦;张振昶;姜华;田亮;袁博;
WANG Jin-hai;DU Xiao-zheng;ZHANG Ting-zhuo;LI Xing-lan;ZHAO Min;BAO Ying-cun;YAN Qi;ZHANG Zhen-chang;JIANG Hua;TIAN Liang;YUAN Bo;Department of Traditional Chinese Medicine,Second Hospital of Lanzhou University;School of Acupuncture-moxibustion and Massage,Gansu University of Traditional Chinese Medicine;Department of Neurology,Second Hospital of Lanzhou University;

• 1:兰州大学第二医院中医科
• 2:甘肃中医药大学针灸推拿学院
• 3:兰州大学第二医院神经内科

摘要(Abstract)：

目的:观察头针对局灶性脑缺血大鼠海马旁回白介素(IL)-10mRNA、IL-6mRNA及肿瘤坏死因子(TNF)-α表达的影响,探讨头针治疗急性缺血性脑血管病的分子生物学机制。方法:SD大鼠随机分为正常组、模型组、药物组及头针组,每组16只。采用线栓法建立急性局灶性脑缺血大鼠模型。药物组予二硫代氨基甲酸吡咯烷100mg·kg^(-1)·d(-1)·d^(-1)腹腔注射,1次/d,共7d;头针组于双侧顶颞前斜线行快速捻转透刺治疗,留针20min,1次/d,共7d。干预前后对各组大鼠进行神经功能缺损评分(NDS)和神经行为学评分(NS)。荧光定量PCR法检测大鼠海马旁回IL-10mRNA、IL-6mRNA的表达,免疫组化法检测海马旁回TNF-α的表达。结果:与正常组比较,模型组大鼠NDS和NS明显升高(P<0.01),海马旁回IL-10mRNA、IL-6mRNA、TNF-α表达明显增高(P<0.01)。与本组治疗前比较,头针组和药物组大鼠NDS和NS明显降低(P<0.01)。治疗后与模型组比较,头针组和药物组大鼠NDS和NS明显降低(P<0.05),海马旁回IL-10mRNA表达明显增高(P<0.05),IL-6mRNA、TNF-α表达明显降低(P<0.05)。结论:促进缺血后海马旁回IL-10的表达,从而抑制IL-6、TNF-α等促炎细胞因子的表达是头针治疗急性缺血性脑血管病的分子机制之一。
Objective To observe the effect of scalp-acupuncture intervention on the expression of Interleukin(IL)-10 mRNA,IL-6 mRNA and tumor necrosis factor(TNF)-αin the parahippocampal gyrus of cerebral ischemia(CI)rats,so as to explore its molecular mechanisms underlying improvement of CI.Methods A total of 64 male SD rats were randomized into normal control,model,medication and scalp-acupuncture groups(n=16 rats in each group).The focal CI model was established by middle cerebral artery occlusion.Intraperitoneal injection of Ammonium Pyrrolidine Dithiocarbamate(100 mg·kg(-1)腹腔注射,1次/d,共7d;头针组于双侧顶颞前斜线行快速捻转透刺治疗,留针20min,1次/d,共7d。干预前后对各组大鼠进行神经功能缺损评分(NDS)和神经行为学评分(NS)。荧光定量PCR法检测大鼠海马旁回IL-10mRNA、IL-6mRNA的表达,免疫组化法检测海马旁回TNF-α的表达。结果:与正常组比较,模型组大鼠NDS和NS明显升高(P<0.01),海马旁回IL-10mRNA、IL-6mRNA、TNF-α表达明显增高(P<0.01)。与本组治疗前比较,头针组和药物组大鼠NDS和NS明显降低(P<0.01)。治疗后与模型组比较,头针组和药物组大鼠NDS和NS明显降低(P<0.05),海马旁回IL-10mRNA表达明显增高(P<0.05),IL-6mRNA、TNF-α表达明显降低(P<0.05)。结论:促进缺血后海马旁回IL-10的表达,从而抑制IL-6、TNF-α等促炎细胞因子的表达是头针治疗急性缺血性脑血管病的分子机制之一。
Objective To observe the effect of scalp-acupuncture intervention on the expression of Interleukin(IL)-10 mRNA,IL-6 mRNA and tumor necrosis factor(TNF)-αin the parahippocampal gyrus of cerebral ischemia(CI)rats,so as to explore its molecular mechanisms underlying improvement of CI.Methods A total of 64 male SD rats were randomized into normal control,model,medication and scalp-acupuncture groups(n=16 rats in each group).The focal CI model was established by middle cerebral artery occlusion.Intraperitoneal injection of Ammonium Pyrrolidine Dithiocarbamate(100 mg·kg^(-1)·d(-1)·d^(-1))was administrated for rats in the medication group,once a day for 7 days.For rats of the scalp-acupuncture group,the acupuncture needles were rapidly inserted into bilateral Dingnieqianxiexian(MS6),followed by twirling the needles at 100 cycles/min for 1 min,once again every 10 min during 20 min'needle retention.The treatment was conducted once a day for 7 days.The neurologic deficit score(0-4 points,impaired consciousness,death,etc.)and neurological function score(motor,sensory and sensory tests,0-10 points)were assessed according to Longa's(1989)and Schabitz's(2004)methods,respectively.The expression levels of IL-10 mRNA and IL-6 mRNA were detected by fluorescence quantitative PCR,and the expression of TNF-αwas detected by immunohistochemistry.Results After modeling,the neurologic deficit and neurological function scores and the expression levels of IL-10 mRNA,IL-6 mRNA and TNF-αprotein in the parahippocampus were significantly increased in the model group than in the normal control group(P<0.01).Following the intervention,the neurologic deficit and neurological function scores as well as IL-6 mRNA and TNF-αprotein expression were significantly down-regulated in both scalp-acupuncture and medication groups(P<0.05),and the expression of IL-10 mRNA was obviously increased(P<0.05)relevant to the model group.Conclusion Scalp-acupuncture can improve neurologic function in CI rats,which is related to its effects in up-regulating the expression of IL-10,then down-regulating the expression of IL-6 and TNF-α(reducing inflammatory response)in the parahippocampal gyrus.

关键词(KeyWords)： 头针;急性缺血性脑血管病;海马旁回;白介素-10mRNA;白介素-6mRNA;肿瘤坏死因子-α
Scalp-acupuncture;Acute cerebral ischemia;Parahippocampus;IL-10mRNA;IL-6mRNA;Tumor necrosis factor-α(TNF-α)

Abstract:

Keywords:

基金项目(Foundation): 兰州大学第二医院院内博士科研基金(No.2015-1-26)

作者(Authors): 王金海;杜小正;张婷卓;李兴兰;赵敏;鲍英存;鄢琦;张振昶;姜华;田亮;袁博;
WANG Jin-hai;DU Xiao-zheng;ZHANG Ting-zhuo;LI Xing-lan;ZHAO Min;BAO Ying-cun;YAN Qi;ZHANG Zhen-chang;JIANG Hua;TIAN Liang;YUAN Bo;Department of Traditional Chinese Medicine,Second Hospital of Lanzhou University;School of Acupuncture-moxibustion and Massage,Gansu University of Traditional Chinese Medicine;Department of Neurology,Second Hospital of Lanzhou University;

DOI: 10.13702/j.1000-0607.180046

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