田亮;王金海;赵敏;鲍英存;商俊芳;鄢琦;张振昶;杜小正;姜华;孙润洁;袁博;张星华;张婷卓;李兴兰;
TIAN Liang;WANG Jin-hai;ZHAO Min;BAO Ying-cun;SHANG Jun-fang;YAN Qi;ZHANG Zhen-chang;DU Xiao-zheng;JIANG Hua;SUN Run-jie;YUAN Bo;ZHANG Xing-hua;ZHANG Ting-zhuo;LI Xing-lan;School of Acu-moxibustion and Massage,Gansu University of Traditional Chinese Medicine;Department of Traditional Chinese Medicine,the Second Hospital of Lanzhou University;Department of Neurology,the Second Hospital of Lanzhou University;School of Medicine and Technology,Gansu University of Traditional Chinese Medicine;

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

摘要(Abstract)：

目的:观察头穴透刺对局灶性脑缺血大鼠海马CA 1区神经细胞膜酸感受离子通道(ASIC)1a、2b表达的影响,探讨头针治疗缺血性中风病的作用机制。方法:SD大鼠随机分为正常组、模型组、头针组和阿米洛利组,每组8只。采用大脑中动脉线栓法复制局灶性脑缺血模型。头针组于双侧顶颞前斜线和顶颞后斜线行快速捻转透刺治疗,1次/d,共7d;阿米洛利组以阿米洛利溶液(5mL/kg,0.045mg/mL)灌胃,2次/d,共7d。于造模成功后1h及干预结束后对各组大鼠进行神经功能缺损评分。干预结束后处死大鼠,快速分离海马组织,免疫组化法检测海马CA 1区神经细胞膜ASIC 1a、ASIC 2b的表达,流式细胞仪检测海马神经元细胞内Ca2+浓度。结果:与正常组比较,模型组神经功能缺损评分显著增高(P<0.01),海马CA 1区神经细胞膜ASIC 1a和ASIC 2b表达显著增高(P<0.01),神经细胞内Ca2+浓度显著增高(P<0.01)。与模型组比较,头针组和阿米洛利组神经功能缺损评分明显降低(P<0.01,P<0.05),且头针组优于阿米洛利组(P<0.05);头针组和阿米洛利组大鼠海马CA 1区神经细胞膜ASIC 1a和ASIC 2b表达均明显降低(P<0.05,P<0.01),细胞内Ca2+浓度明显降低(P<0.01)。结论:下调神经元细胞膜ASIC1a、ASIC 2b表达,进而降低大鼠海马神经细胞内Ca2+浓度,抑制神经细胞凋亡可能是头针治疗缺血性中风病的作用机制之一。
Objective To observe the influence of scalp-acupuncture on the expression of acid-sensing ion channels(ASICs)1aand 2bof hippocampal CA 1region in cerebral ischemia(CI)rats,so as to investigate its mechanism underlying improvement of ischemic stroke.Methods Thirty-two male SD rats were randomly allocated to normal control,model,scalp-acupuncture and Amiloride group(n=8in each group).The model of focal CI was established by middle cerebral artery occlusion(MCAO).Scalp acupuncture stimulation was applied to bilateral Dingnieqianxiexian(MS 6)and Dingniehouxiexian(MS 7),once daily for 7days.Rats of the Amiloride group were fed with Amiloride solution,twice a day for 7days,and those of the normal control and model groups were grabbled and fixed in the same way with the acupuncture and Amiloride groups.The neurological deficit score was given according to Longa's method.The expression of hippocampal ASIC 1aand ASIC 2bwas detected by immunohistochemistry,and the Ca2+concentration in the hippocampal tissue assayed using flowing cytometry.Results After the intervention,the neurological deficit score of both the scalp-acupuncture and Amiloride groups were significantly decreased in comparison with pre-treatment(P<0.01),and the effect of scalp-acupuncture was markedly superior to that of Amiloride in reducing neurological deficit score(P<0.05).The expression of ASIC 1aand ASIC 2bin the hippocampal CA 1region and hippocampal Ca2+concentration were significantly up-regulated in the model group compared with the normal control group(P<0.01),and obviously down-regulated in both scalp-acupuncture and Amiloride groups(P<0.01,P<0.05),without significant differences between the two treatment groups in the ASIC 1aand ASIC 2bexpression and Ca2+concentration(P>0.05).Conclusion Scalp-acupuncture stimulation can improve neurological function in CI rats,which may be related to its effects in suppressing the increased expression of hippocampal ASIC 1aand ASIC 2bproteins and in reducing calcium overload in hippocampal neurocytes.

关键词(KeyWords)： 头针;局灶性脑缺血;海马CA 1区;酸感受离子通道1a;酸感受离子通道2b;钙离子浓度
Scalp acupuncture;Focal cerebral ischemia;Hippocampal CA 1region;Acid-sensing ion channels(ASIC)1a;ASIC 2b;Calcium ion concentration

Abstract:

Keywords:

基金项目(Foundation): 甘肃省中医药管理局资助项目(No.GZK-2015-54);; 兰州大学第二医院院内中医药课题(No.YJzy2013-39);; 2015年甘肃中医药大学研究生创新基金资助项目(No.2016CX 27)

作者(Authors): 田亮;王金海;赵敏;鲍英存;商俊芳;鄢琦;张振昶;杜小正;姜华;孙润洁;袁博;张星华;张婷卓;李兴兰;
TIAN Liang;WANG Jin-hai;ZHAO Min;BAO Ying-cun;SHANG Jun-fang;YAN Qi;ZHANG Zhen-chang;DU Xiao-zheng;JIANG Hua;SUN Run-jie;YUAN Bo;ZHANG Xing-hua;ZHANG Ting-zhuo;LI Xing-lan;School of Acu-moxibustion and Massage,Gansu University of Traditional Chinese Medicine;Department of Traditional Chinese Medicine,the Second Hospital of Lanzhou University;Department of Neurology,the Second Hospital of Lanzhou University;School of Medicine and Technology,Gansu University of Traditional Chinese Medicine;

DOI: 10.13702/j.1000-0607.2016.05.010

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