电针预处理对肢体缺血再灌注大鼠生存、脑损伤及认知功能的影响  

Effects of electroacupuncture pretreatment on survival,brain injury and cognitive function in rats after limb ischemia/reperfusion

作　　者：陈烨[1] 周军[2] 袁婉丽[1] 莫利群[2] 胡节惠[1] 

机构地区：[1]泸州医学院附属医院中医科,四川泸州646000 [2]泸州医学院附属医院麻醉科,四川泸州646000

出　　处：《中国病理生理杂志》2013年第3期469-475,共7页Chinese Journal of Pathophysiology

基　　金：泸州医学院附属医院人才基金资助项目(No.2011-43-43)

摘　　要：目的:观察电针(EA)预处理对肢体缺血再灌注(LI/R)大鼠生存情况、脑损伤及认知功能的影响,探讨相关机制。方法:清洁级、健康成年雄性SD大鼠132只,体重255～300 g,采用随机数字表法,将其随机分为3组:假手术(sham)组、LI/R组和LI/R+EA预处理(LI/R+EA)组。LI/R组采用动脉夹夹闭双后肢股动脉3 h,建立LI/R模型;sham组只暴露股动脉,不夹闭;LI/R+EA组于模型制备前14 d行电针治疗,针刺穴位为"百会"、"足三里"及"血海"。观察各组大鼠7 d生存率;于再灌注48 h,用Morris水迷宫方法测试大鼠认知功能的变化;其余大鼠于再灌注48 h后处死取材,用干湿重法测定脑含水量;免疫组织化学法测定小胶质细胞标志物Iba1;采用Western blotting检测cleaved caspase-3的表达水平;原位末端标记(TUNEL)法检测神经细胞凋亡并计算凋亡指数;光镜下观察海马病理结构变化;化学法测定活性氧(ROS)、丙二醛(MDA)含量及髓过氧化物酶(MPO)、超氧化物歧化酶(SOD)活性。结果:与sham组比较,LI/R组及LI/R+EA组大鼠7 d生存率下降,潜伏期和游泳距离增加,穿越平台次数减少,海马Iba1阳性细胞数增加,cleaved caspase-3蛋白表达量增加,凋亡指数明显增加,神经元减少,ROS、MDA含量及MPO活性增加,SOD活性降低(P<0.05或P<0.01);与LI/R组相比,LI/R+EA组上述指标得到明显改善(P<0.05或P<0.01)。结论:电针预处理能提高LI/R大鼠生存率,减轻脑损伤及改善大鼠认知功能障碍,其机制可能与抑制小胶质细胞激活、减轻氧化应激损伤有关。AIM： To investigate the effects of electroacupuncture （EA） pretreatment on survival, brain injury and cognitive function in rats after limb ischemia/reperfusion （LI/R）. METHODS： One hundred and thirty-two healthy male SD rats weighing 255 300 g were randomly divided into 3 groups （ n =44 each） ： sham operation group （ sham）, LI/ R group and LI/R plus EA pretreatment （IL/R ＋ EA） group. The IZ/R model was made by a method that the bilateral femoral arteries were occluded for 3 h with atraumatic microclips followed by 48 h of reperfusion. In sham group, sham op- eration was performed. The EA pretreatment was conducted twice a day for 14 d prior to the LI/R event. EA pretreatment included the following acupoints： Baihui （GV20）, Zusanli （ST36） and Xuehai （SP10）. The survival rate within 7 d fol- lowing LIZR was calculated. The changes of cognitive function were detected 48 h after reperfusion using Morris water maze test. The cerebral water content was determined by detecting the wet and dry weight. Microglial cells were evaluated follow- ing immunolabeling of Ibal （ a marker of microglia）. The protein level of cleaved caspase-3 in the hippocampus was meas- ured by Western blotting. The neuronal apoptosis was detected using TUNEL method. Meanwhile, the changes of pathologi- cal structure in hippocampus were observed under light microscope. The activity of MPO and SOD, and the content of ROS and MDA were also investigated. RESULTS： Compared with sham group, the Ibal positive cells, the protein level of cleaved caspase-3, the apoptotic index, and the levels of ROS/MDA and MPO activity in LI/R and LI/R ＋ EA groups in- creased significantly. The normal hippocampal neurons reduced, and SOD activity decreased significantly in hippocampus.The survival rates of the rats within 7 d decreased, the latency and swimming distance increased, and the number of cross- ing the platform reduced. Compared with LI/R group, the above indexes in LI/R ＋ EA group were markedly improved. CO

关 键 词：电针 肢体再灌注损伤 认知功能 小神经胶质细胞 氧化性应激 

分 类 号：R363[医药卫生—病理学]