不同时程电针预处理对脑缺血再灌注大鼠血脑屏障水通道蛋白-4及紧密连接蛋白-5的影响  

The impact of electroacupuncture on the expression of AQP4 and CLN5 in the blood-brain barrier after cerebral ischemia and reperfusion

作　　者：林咸明[1] 李金霞[2] 姚旭[1] 

机构地区：[1]浙江中医药大学第三临床医学院,杭州310053 [2]浙江省湖州市中医院针灸科

出　　处：《中华物理医学与康复杂志》2015年第7期498-502,共5页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：浙江省自然科学基金项目（Y2111260）

摘　　要：目的观察不同时程电针（取穴百会、水沟）预处理对继发脑缺血再灌注大鼠血脑屏障水通道蛋白-4（AQP4）及紧密连接蛋白-5（CLN5）表达的影响。 方法采用随机数字表法将64只雄性SD大鼠分为模型组、假手术组、电针预处理7d组及电针预处理15d组，电针预处理7d组及电针预处理15d组分别给予持续7d、15d的电针（取穴百会、水沟）预处理。待上述预处理结束后，参照Longa改良线栓法将模型组、电针预处理7d组及电针预处理15d组大鼠制成单侧大脑中动脉栓塞（MCAO）模型，假手术组大鼠术中仅分离颈总动脉，不栓塞中动脉血流。各组大鼠于大脑中动脉栓塞90min后进行再灌注。于制模24h后采用免疫组化法、荧光定量PCR法检测各组大鼠血脑屏障AQP4、CLN5阳性细胞及其mRNA表达情况。 结果与假手术组比较，模型组及各电针预处理组大鼠血脑屏障AQP4阳性细胞及其mRNA表达均明显上调（P〈0.05），CLN5阳性细胞及其mRNA表达均明显下调（P〈0.05）；与模型组比较，电针预处理7d组及15d组AQP4阳性细胞及其mRNA表达均明显下调（P〈0.05），CLN5阳性细胞及其mRNA表达均明显上调（P〈0.05）；与电针预处理7d组比较，电针预处理15d组AQP4阳性细胞数［（25.15±0.55）个/每高倍镜视野］及其mRNA表达（1.16±0.04）下调幅度、CLN5阳性细胞数［（62.88±0.61）个/每高倍镜视野］及其mRNA表达（0.80±0.03）上调幅度均更显著（P〈0.05）。 结论不同时程电针（取穴百会、水沟）预处理可抑制脑缺血再灌注大鼠血脑屏障AQP4阳性细胞及其mRNA表达，促进CLN5阳性细胞及其mRNA表达；以电针预处理15d对脑缺血再灌注大鼠血脑屏障的保护作用相对较好；电针预处理诱导大鼠脑缺血耐受可能与调节脑缺血再灌后血脑屏障AQP4、CLN5及其mRNA表达有关。Objective To observe the effect of electroacupuncture （EA） pretreatment of different durations on the expressions of AQP4 and CLN5 in the blood-brain barrier after cerebral isehemia and reperfusion Methods Sixty-four male Sprague-Dawley （SD） rats were randomly divided into a model group, a sham operation group, a 7- day EA-pretreatment group （EAP-7 group） and a 15-day EA-pretreatment group （EAP-15 group） , each of 16. After EA-pretreatment on the baihui and shuigou acupoints for 7 days or 15 days, a model of unilateral middle cerebral artery embolism （MCAO） model was induced in the rats of the model, EAP-7 and EAP-15 group using a modified Longa method. In the sham operation group the carotid artery was separated without middle cerebral artery embolism. The reperfusion was begun 90 min after the MCAO modeling. Immunohistochemical methods and fluorescence quantitative PCR were applied to detect the expression of AQP4, CLN5 and their mRNAs expression in the blood brain barrier （BBB） 24h after the operation. Results Compared with the sham operation group, the expression of AQP4 positive cells and AQP4 mRNA in the BBB in the other three groups had increased significantly, while the expression of CLN5 positive cells and CLN5 mRNA was significantly less. Compared with the model group, the expression of AQP4 positive cells and AQP4 mRNA in the EAP-7 and EAP-15 groups was significantly reduced, while the expression of CLN5 positive cells and CLN5 mRNA was significantly increased. Moreover, the expression of AQP4 positive cells and AQP4 mRNA in the EAP-15 group were significantly higher than in the EAP-7 group, while the expression of CLN5 positive cells and CLN5 mRNA were significantly lower. Conclusions EA-pretreat-ment on the baihui and shuigou acupoints can restrain the expression of AQP4-positive ceils and AQP4 mRNA and promote that of CLN5-positive cells and CLN5 mRNA in the BBB after cerebral ischemia and reperfusion, at least in rats. The BBB protection effect is better when the EA-pretreat

关 键 词：电针预处理 脑缺血再灌注 血脑屏障 水通道蛋白-4 紧密连接蛋白-5 

分 类 号：R245[医药卫生—针灸推拿学]