文献类型：期刊文献

中文题名：温通针法抑制神经元钙超载治疗血管性痴呆的机制探讨

英文题名：Preventing role of warmth-promotion needling in reducing intracellular Ca^2+ overload of in vitro hippocampal neuronal cells of vascular dementia rats

作者：杨晓波[1,2];王金海[3];安惠琴[1];刘丁龙[1];关姝明[1];崔道宁[1];赵彬元[1];张小丽[1];舒畅[4]

第一作者：杨晓波

机构：[1]甘肃中医药大学针灸推拿学院,兰州730000;[2]中国医科大学航空总医院中医科,北京100021;[3]兰州大学第二医院中医科,兰州730030;[4]甘肃中医药大学科研实验中心,兰州730000

第一机构：甘肃中医药大学针灸推拿学院

年份：2020

卷号：45

期号：6

起止页码：454

中文期刊名：针刺研究

外文期刊名：Acupuncture Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；PubMed;

基金：国家自然科学基金项目(No.81360563)。

语种：中文

中文关键词：血管性痴呆;针刺;温通针法;L-型钙通道电流;细胞内Ca2+含量;全细胞膜片钳技术;海马CA1区

外文关键词：Vascular dementia;Acupuncture;Warmth-promotion needling;L-type calcium channel;Intracellular calcium content;Whole cell patch clamp recording;Hippocampal CA1 region

摘要：目的:观察温通针法对血管性痴呆(VD)大鼠海马神经元L-型钙通道电流(Ica.L)、神经元内Ca^2+含量及脑组织形态学的影响,探讨温通针法治疗VD的机制。方法:SD大鼠按随机数字表法分为正常组、模型组、药物组、温通针法组、平补平泻组和捻转针法组,每组18只。采用反复夹闭双侧颈总动脉-再灌注法复制VD模型。药物组给予尼莫地平溶液灌胃,每日2次,共14 d;温通针法组、平补平泻组和捻转针法组于"大椎""百会""水沟"3穴分别施以温通针法、平补平泻法和捻转针法针刺,每次留针30 min,每日1次,共治疗14 d。采用全细胞膜片钳技术观察各组大鼠海马神经元Ica.L的变化,激光共聚焦扫描显微镜观察海马CA1区神经元内Ca^2+含量的变化,HE染色法观察大鼠海马CA1区组织形态学改变。结果:与正常组比较,模型组大鼠海马神经元Ica.L的电流密度明显增加(P<0.05),Ica.L的I-V曲线明显下移,神经元内Ca^2+含量明显升高(P<0.05),海马CA1区神经元数量明显减少,细胞间隙增宽,胞体肿胀、变性,核固缩,排列紊乱,胶质细胞数量增多。与模型组比较,药物组、温通针法组、平补平泻组、捻转针法组海马神经元Ica.L的电流密度明显降低(P<0.05),Ica.L的I-V曲线明显上移,神经元内Ca^2+含量明显降低(P<0.05),且温通针法组变化比平补平泻组和捻转针法组更明显(P<0.05);海马CA1区神经元数量明显增多,核固缩、变性减轻,胶质细胞增生不明显,形态较模型组明显改善。结论:温通针法可显著降低VD大鼠海马CA1区神经元Ica.L的电流密度及神经元内Ca^2+含量,抑制神经元钙超载,拮抗脑缺血再灌注后继发神经元的损伤,温通针法治疗VD具有针刺效应相对特异性。
Objective To investigate the underlying mechanisms of warming-promotion needling(WPN)for vascular dementia(VD)by observing its effect on the expression of L-type calcium channel current(Ica.L),changes of intracellular[Ca2+]i in neurons and pathomorphological changes of brain in VD rats.Methods One hundred and eight male SD rats were randomized into normal control,model,medication,WPN,uniform reinforcing-reducing needling(URRN),and needle-twirling groups(n=18 in each group).The VD model was established by repeated occlusion and reperfusion of the bilateral common carotid arteries.Rats of the medication group were treated by gavage of Nimodipine twice a day for 14 days.For rats of the 3 acupuncture groups,WPN or URRN or needle-twirling was applied to"Dazhui"(GV14),"Baihui"(GV20)and"Shuigou"(GV26)for 30 min,once a day for 14 days.Changes of Ica.L in hippocampal neurons were observed by whole-cell patch clamp technique,and levels of[Ca2+]i in neuronal cells of living brain slice of hippocampal CA1 area were observed by laser confocal microscope.Histopathological changes of hippocampal CA1 area were observed under light microscope after H.E.stain.Results Compared with the normal control group,the current density of Ica.L,and the intracellular[Ca2+]i levels were significantly increased(P<0.05),and the I-V curves of Ica.L was apparently shifted down in the model group.In comparison with the model group,the current density of Ica.L was obviously reduced(P<0.05),the I-V curves of Ica.L shifted up and the intracellular Ca2+content notably decreased in the medication,WPN,URRN and needle-twirling groups(P<0.05).The effect of WPN was significantly superior to those of URRN and needle-twiiling(P<0.05).H.E.stain showed a reduction in the number of hippocampal neurons,dilation of intercellular space,swelling of cell body,karyopyknosis,disordered arrangement and increased number of gliacytes in the model group,which was apparently milder in the medication and 3 acupuncture groups(the WPN group in particular).Conclusion The WPN can reduce Ica.L current density and[Ca2+]i content of hippocampal neurons in brain slices of VD rats,which may contribute to its effect in relieving VD by suppressing calcium overload.