耳穴经皮电刺激对颞叶癫痫大鼠癫痫发作频率与海马区胶质细胞活性及炎性因子的影响  

Transcutaneous Electrostimulation of Auricular Otopoints Reduces Epileptic Attack Possibly by Suppressing Hippocampal Gliocyte Proliferation and Regulating IL-6 and IL-10 Expression in Chronic Temporal Lobe Epilepsy Rats

作　　者：乔丽娜[1] 杨海龙[1,2] 谭连红[1] 杨娇娇[1,3] 高巧玲[1,3] 周胜红[4] 张永臣[2] 杨永升[1] 

机构地区：[1]中国中医科学院针灸研究所生物化学与分子生物学研究室,北京100700 [2]山东中医药大学针灸推拿学院,济南250014 [3]长春中医药大学针灸推拿学院,长春130117 [4]山东省中医药研究院,济南250014

出　　处：《针刺研究》2017年第3期189-196,共8页Acupuncture Research

基　　金：国家自然科学基金面上项目(No.81273833);国家自然科学基金青年基金项目(No.81303055);国家重点基础研究发展计划"973"项目(No.2014CB 543103;2013CB 531904)

摘　　要：目的:观察经皮电刺激耳迷走神经支配的耳穴对颞叶癫痫大鼠自发癫痫发作频率、海马区胶质细胞活性及促炎细胞因子白细胞介素(IL)-6、抗炎细胞因子IL-10表达的影响,探讨耳穴刺激对颞叶癫痫的缓解效应及其可能的细胞与分子机制。方法:SD大鼠随机分为正常组、模型组、治疗组,每组12只。采用氯化锂-匹罗卡品多次小剂量重复注射制备慢性颞叶癫痫大鼠模型。对治疗组耳甲腔内耳穴"心""肺""皮质下"进行经皮电刺激干预,每日1次,每次20min,治疗6周。用长时程视频系统监测大鼠的癫痫发作情况,治疗后用免疫荧光与Real-time PCR方法观察癫痫大鼠海马区小胶质细胞标记物Iba 1、星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)及IL-6、IL-10的表达情况。结果:模型组大鼠癫痫自发频率随时间延长显著增加,治疗组大鼠的自发癫痫频率较模型组显著降低(P<0.05)。与正常组比较,模型组大鼠海马CA 1、CA 3区Iba 1、GFAP、IL-6免疫阳性细胞数明显增多(P<0.05),IL-6mRNA表达水平显著上调(P<0.05)。与模型组比较,治疗组大鼠海马CA 1、CA 3区Iba 1、GFAP、IL-6免疫阳性细胞数明显减少(P<0.05),IL-6mRNA表达水平显著下调(P<0.05),IL-10阳性细胞数及mRNA表达水平显著上调(P<0.05)。结论:耳迷走神经支配的耳穴经皮电刺激干预能显著降低慢性颞叶癫痫大鼠的癫痫发作频率,此效应可能与下调海马区胶质细胞活性,下调海马内促炎细胞因子IL-6,上调抗炎细胞因子IL-10的蛋白及基因表达水平密切相关。Objective To observe the effect of transcutaneous otopoint electrostimulaiton（TCOES）on seizure frequency,immunoreactivity of hippocampal gliocytes and expression of proinflammatory cytokine interleukin-6（IL-6）and anti-inflammatory cytokine IL-10 in chronic temporal lobe epilepsy（CTLE）rats,so as to investigate its antiepileptic mechanism.Methods Thirtysix SD rats were randomly divided into control,model and TCOES groups（n=12in each group）.The CTLE model was established by intraperitoneal injection（i.p.i.）of lithium chloride（127.2mg/kg）,scopolamine（1mg/kg,20 hafter the 1st injection）and pilocarpine（10mg/kg,30 min after scopolamine injection）.Rats of the control group were treated by i.p.i.of normal saline.TCOES（1mA,20Hz）was applied to bilateral otopoint＂Heart＂-＂Lung＂-＂Subcortex＂region for 20 min,once daily for 6weeks.The epileptic attack was observed by a video monitoring system.The numbers of ionized calcium-binding adapter molecule-1（Iba 1）-labeled microgliacytes and glial fibrillary acidic protein（GFAP）-labeled astrocytes in the CA 1and CA 3regions of hippocampus were counted under light microscope after immunostaining,and the expression levels of hippocampal IL-6and IL-10 proteins and genes were determined by immunofluorescence and quantitative real-time PCR,respectively.Results After TCOES intervention,the seizure frequency was significantly decreased in comparison with pre-treatment（P〈0.05）,modeling-induced dramatic increase of the numbers of microgliacytes and astrocytes,IL-6immunoactivity in the hippocampal CA 1and CA 3regions,and IL-6mRNA expression in the hippocampus were significantly suppressed（P〈0.05）,and hippo-campal IL-10 immunoactivity and mRNA expression were considerably up-regulated in comparison with the model group（P〈0.05）.Conclusion TCOES intervention has an antiepileptic effect in CTLE rats,which may be associated with its effects in suppressing gliocyte proliferation,suppressing the expression of proinflammatory cytoki

关 键 词：颞叶癫痫 经皮耳穴电刺激 海马 胶质细胞 炎性因子 

分 类 号：R245.97[医药卫生—针灸推拿学]