电针预处理对脑缺血再灌注损伤大鼠皮质神经细胞线粒体融合蛋白2表达的影响  

EFFECT OF ELECTROACUPUNCTURE PRECONDITIONING ON THE EXPRESSION OF MITOCHONDRIAL FUSION PROTEIN 2 IN CORTICAL NERVE CELLS OF RATS WITH CEREBRAL ISCHEMIA/REPERFUSION INJURY

作　　者：丁娜[1,2] 袁阳 王明山 张高峰[1] 陈怀龙 时飞[1] 孙晓鹏[1] DING Na;YUAN Yang;WANG Mingshan;ZHANG Gaofeng;CHEN Huailong;SHI Fei;SUN Xiaopeng(Department of Anesthesiology,Affiliated Qingdao Municipal Hospital of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属青岛市市立医院麻醉科,山东青岛266071 [2]青岛市西海岸新区区立医院麻醉科

出　　处：《精准医学杂志》2022年第6期550-553,共4页Journal of Precision Medicine

基　　金：青岛市科技民生项目(19-6-1-50-nsh)。

摘　　要：目的探讨电针预处理对脑缺血再灌注(I/R)损伤大鼠皮质神经细胞线粒体融合蛋白2(Mfn2)表达的影响。方法健康雄性SD大鼠93只,随机分为假手术组(S组)、I/R组、电针预处理组(E组)(n=31)。S组仅分离大鼠颈部血管,不进行阻闭;I/R组采用线栓法制备大脑中动脉I/R损伤模型;E组于造模前连续5 d给予电针刺激百会穴,最后1次针刺结束24 h后同I/R组处理。于再灌注后6、24、48 h按Longa评分法对各组大鼠进行神经功能缺损评分,取大鼠大脑皮质缺血半暗区组织,采用Tunel法测定神经细胞凋亡率,电镜下观察神经细胞线粒体超微结构,分离神经细胞线粒体和胞浆后采用Western blot法检测线粒体上Mfn2及胞浆中Cyt C蛋白的表达水平。结果分组对神经功能缺损评分有明显影响(F=235.53,P<0.05)。时间、分组、时间与分组交互作用对神经细胞凋亡率、线粒体上Mfn2及胞浆中Cyt C表达水平均有明显影响(F=13.71~2019.71,P<0.05);与S组比较,I/R组和E组再灌注后6、24、48 h神经功能缺损评分、神经细胞凋亡率及胞浆中Cyt C水平均显著升高,线粒体上Mfn2水平显著下降(P<0.01);与S组相比,I/R组和E组再灌注后24 h线粒体分裂,膜间隙肿胀,线粒体嵴模糊或消失;与I/R组比较,E组再灌注后6、24、48 h神经功能缺损评分、神经细胞凋亡率及胞浆中Cyt C水平显著降低,线粒体上Mfn2水平显著升高(P<0.01),再灌注后24 h线粒体超微结构改变减轻。结论电针预处理可减轻大鼠脑I/R损伤,其机制可能与上调大脑皮质缺血半暗区线粒体上Mfn2表达,增强线粒体融合有关。Objective To investigate the effect of electroacupuncture(EA) preconditioning on the expression of mitochondrial fusion protein 2(Mfn2) in cortical nerve cell of rats with cerebral ischemia-reperfusion(I/R) injury. Methods A total of 93 healthy male Sprague-Dawley rats were randomly divided into sham-operation group(S group), I/R group, and EA preconditioning group(E group), with 31 rats in each group. For the S group, cervical vessels were separated without occlusion;for the I/R group, the suture method was used to establish a model of middle cerebral artery I/R injury;for the E group, the rats were given EA at Baihui acupoint for 5 consecutive days before modeling, followed by the same treatment as the I/R group at 24 h after the last EA stimulation. At 6, 24, and 48 h after reperfusion, the Longa scoring method was used to determine the neurological deficit score of rats in each group;after tissue samples were collected from the ischemic penumbra of the cerebral cortex, the Tunel method was used to measure the apoptosis rate of nerve cells, and mitochondrial ultrastructure in nerve cells was observed under an electron microscope;after isolation of mitochondria and cytoplasm in nerve cells, Western blot was used to measure the expression levels of mitochondrial Mfn2 and cytoplasmic Cyt C. Results Group had a significant effect on neurological deficit score(F=235.53,P<0.05), and time, group, and time-group interaction had a significant effect on the apoptosis rate of nerve cells and the expression levels of mitochondrial Mfn2 and cytoplasmic Cyt C(F=13.71-2019.71,P<0.05). Compared with the S group, the I/R group and the E group had significant increase in neurological deficit score, the apoptosis rate of nerve cells, and the level of cytoplasmic Cyt C and a significant reduction in the level of mitochondrial Mfn2 at 6, 24, and 48 h after reperfusion(P<0.01). Compared with the S group, the I/R group and the E group had mitochondrial fission, intermembrane swelling, and vague or disappeared mitochondrial cristae.

关 键 词：电刺激疗法 再灌注损伤 大脑皮质 疾病模型 动物 线粒体蛋白质类 细胞色素C类 脑保护 

分 类 号：R454.1[医药卫生—治疗学] R743.31[医药卫生—临床医学]