电针通过抑制AQP4表达减轻局灶脑缺血/再灌注大鼠脑水肿损伤及星形胶质细胞激活  

Electroacupuncture attenuate cerebral edema injury and astrocyte activation by suppressing the expression of aquaporin AQP4 in focal cerebral ischemia/reperfusion rat

作　　者：卞炜 秦文熠[2] BIAN Wei;QIN Wenyi(Department of Rehabilitation,Chongqing Yongchuan TCM Hospital 402160,China;Department of Integrated Traditional Chinese and Western Medicine,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆市永川区中医院康复科,402160 [2]重庆医科大学附属第一医院中西医结合科,重庆400016

出　　处：《现代医药卫生》2018年第5期665-668,共4页Journal of Modern Medicine & Health

基　　金：国家自然科学基金资助项目(81403243)

摘　　要：目的观察电针对局灶脑缺血/再灌注大鼠大脑缺血皮质区水通道蛋白4(AQP4)表达的影响,探讨电针减轻脑水肿和星形胶质细胞激活的可能机制。方法将60只SD大鼠分为正常组(NC组)、模型组(t MCAO组)、模型+电针治疗组(EA组)、模型+电针+AQP4特异性拮抗剂TGN-02组(TGN-02组),每组15只。采用线栓法制备大鼠局灶脑缺血/再灌注模型,取大鼠"百会"穴及左侧"四关"穴为电针穴位。对除NC组外的其余各组建立局灶脑缺血/再灌注模型。再灌注后24 h行脑组织含水量检测,Western blotting检测AQP4蛋白表达,实时定量聚合酶链反应(RT-q PCR)检测AQP4、神经胶质纤维酸性蛋白(GFAP)mRNA表达情况,免疫荧光共聚焦观察GFAP标记的星形胶质细胞。结果再灌注后24 h,EA组脑水肿较模型组减轻,TGN-02组脑水肿较EA组明显加重,差异均有统计学意义(P<0.05)。EA组大鼠大脑缺血皮质区AQP4蛋白及mRNA表达较t MCAO组显著降低,TGN-02组GFAP mRNA表达较EA组明显降低,差异均有统计学意义(P<0.05)。荧光共聚焦结果显示,t MCAO组GFAP标记的星形胶质细胞形态增大、肿胀,突起增多、增粗。EA组大脑缺血皮质区星形胶质细胞数量较t MCAO组明显降低,而AQP4作用被阻断后,电针治疗未能减少星形胶质细胞数量。结论电针可通过下调局灶脑缺血/再灌注大鼠大脑缺血皮质区AQP4表达,削弱脑内星形胶质细胞激活,减轻脑水肿损伤。Objective To explore the possible mechanism of electroacupuncture in reducing cerebral edema and astrocyte activation by observing the expression of AQP4in the cerebral ischemic cortex in focal cerebral ischemia/reperfusion rats.Methods Sixty SD rats were randomly divided into normal group(NC group),model group(tMCAO group),model+elec?troacupuncture treatment group(EA group),model+electroacupuncture+AQP4specific antagonist TGN?02group(TGN?02group),15rats in each group.The rat of focal cerebral ischemia/reperfusion model was prepared by suture method.The“Bai?hui”and the“Siguan”were chosen as electroacupuncture points.tMCAO model was established for groups except the NC group.Water content of brain tissue was detected24h after reperfusion,the expression of AQP4protein was detected by Western blot-ting,the expression of AQP4mRNA and astrocyte marker GFAP mRNA were detected by RT?qPCR,GFAP?labeled astrocytes was observed by immunofluorescence confocal microscopy.Results24h after reperfusion,the cerebral edema in the EA group was significantly lower than that in the tMCAO group,the cerebral edema in the TGN?02group was significantly worse than that in the EA group,the differences were statistically significant(P<0.05).The expression of AQP4protein and mRNA in the cerebral ischemic cortex of EA group were significantly lower than those in tMCAO group,the expression of GFAP mRNAin the TGN?02group was significantly lower than that in the EA group,the differences were statistically significant(P<0.05).Fluorescence confocal microscopy showed that the number of GFAP?labeled astrocytes increased in the tMCAO group,andtheir protrusions increased.The number of astrocytes in the cerebral ischemic cortex of EA group was significantly lower than that of tMCAO group,and decreased after blocked by TGN?02.Conclusion Electroacupuncture can reduce the activation of astrocytes and reduce the brain edema by reducing the expression of AQP4in the cerebral ischemic cortex in focal cerebral ischemia/reperfusion rats.

关 键 词：脑缺血 再灌注 电针 脑水肿 水孔蛋白质类 星形细胞 大鼠 sprague-dawley 

分 类 号：R245[医药卫生—针灸推拿学]