“通督调神”电针预处理大鼠缺血半暗带区miR-124和皮质区蛋白的表达  

Expression of microRNA-124 and cortical proteins in ischemic penumbra of rats pretreated with“Tongdu Tiaoshen”electroacupuncture

作　　者：张君宇[1] 张利达 胡滟琦 金子开 罗佛赐 吴晓晴 童婷婷 宋小鸽 韩为[1] Zhang Junyu;Zhang Lida;Hu Yanqi;Jin Zikai;Luo Fuci;Wu Xiaoqing;Tong Tingting;Song Xiaoge;Han Wei(The Second Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230001,Anhui Province,China;Department of Traditional Chinese Medicine,Affiliated Brain Hospital of Guangzhou Medical University,Guangzhou 510370,Guangdong Province,China;College of Acupuncture,Moxibustion and Tuina,Anhui University of Traditional Chinese Medicine,Hefei 230031,Anhui Province,China;Wangjing Hospital,Chinese Academy of Traditional Chinese Medicine,Beijing 100102,China;The Second Hospital of Traditional Chinese Medicine of Jiangsu Province,Nanjing 210017,Jiangsu Province,China;Graduate School of Anhui University of Traditional Chinese Medicine,Hefei 230022,Anhui Province,China;Anhui Acupuncture and Meridian Research Institute,Hefei 230031,Anhui Province,China)

机构地区：[1]安徽中医药大学第二附属医院,安徽省合肥市230001 [2]广州医科大学附属脑科医院中医科,广东省广州市510370 [3]安徽中医药大学针灸推拿学院,安徽省合肥市230031 [4]中国中医科学院望京医院,北京市100102 [5]江苏省第二中医院,江苏省南京市210017 [6]安徽中医药大学研究生院,安徽省合肥市230022 [7]安徽省针灸经络研究所,安徽省合肥市230031

出　　处：《中国组织工程研究》2023年第26期4139-4146,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:“通督调神”电针预处理法是结合传统针刺和现代电刺激的新兴物理治疗方法,既往文献报道其对缺血性脑卒中具有显著治疗作用,但其具体机制尚不明确。目的:探讨“通督调神”电针预处理作用于细胞凋亡,对大鼠缺血半暗带区miR-124和皮质区Notch-1、p-JNK和半胱氨酸天冬氨酸特异性蛋白酶3蛋白相对表达量的影响及其可能的作用机制。方法:将75只雄性SD大鼠随机分为模型组、假手术组、电针预处理组、miR-124抑制剂预处理组和电针+miR-124激动剂预处理组,每组15只。7 d干预期内,电针预处理组取“风府”“百会”“大椎”穴行电针,1次/d,20 min/次;miR-124抑制剂预处理组第7天注射miR-124 antagomir至大鼠侧脑室内;电针+miR-124激动剂预处理组第7天在电针基础上注射miR-124 agomir;模型组及假手术组不予治疗。干预结束后造模,假手术组只做鼠板上固定和钝性分离血管,其余组均采用大脑中动脉闭塞方法并参照Zea Longa线栓法建立大鼠右侧脑缺血再灌注损伤模型。造模完成后24 h应用改良神经损伤程度评分定损,评分结束后使用TUNEL染色法测细胞凋亡,电镜观察大鼠皮质脑区超微结构,Western Blot法检测皮质区蛋白相对表达量,实时荧光定量PCR检测mRNA相对表达量。结果与结论:①与假手术组比较,其余各组大鼠改良神经损伤程度评分均升高(均P<0.01);与模型组比较,miR-124抑制剂预处理组和电针预处理组改良神经损伤程度评分显著降低(均P<0.05);②与假手术组比较,模型组大鼠凋亡细胞数量显著升高(P<0.01);与模型组比较,各预处理组大鼠凋亡细胞数量显著降低(均P<0.01);③模型组细胞结构及内容物缺损较重;电针预处理组细胞结构及内容物轻度缺损;miR-124抑制剂预处理组细胞较完整;电针+miR-124激动剂预处理组细胞结构及内容物存在损伤;④与模型组比较,miR-124抑制剂预处理组和电针BACKG ROUND:"Tongdu Tiooshen" electroacupuncture pretreatment is an eme rging physiotherapy combining traditional acupuncture with modern electrical stimulation,which has been reported to have significant therapeutic effects on ischemic stroke,but its specific mechanism is still unclear.OBJECTIVE:To investigate the effects of Tongdu Tiaoshen" electroacupuncture pretreatment on cell apoptosis,the relative expression of microRNA-124(miR-124) in the ischemic penumbra and Notch-1,p-JNK and cysteinyl-aspartate specific protease-3(caspase-3) proteins in the co rtex of rats and their possible mechanisms of action.METHODS:Seventy-five male Sprague-Dawley rats were randomly divided into five groups(n=15 per group):model group,sham-operated group,electroacupuncture pretreatment group,miR-124 antagomir pretreatment group and electroacupuncture+miR-124 agomir pretreatment group.In the electroacupuncture pretreatment group,20-minute electroacupuncture at Fengchi,Baihui,and Dazhui was performed once a day,for 7 continuous days.In the miR-124 antagomir pretreatment group,miR-124 antagomir was injected into the lateral ventricle of rats on day 7.In the electroacupuncture+miR-124 agomir pretreatment group,miR-124 agomir was injected based on electroacupuncture pretreatment.Model and sham-operated groups were not treated.After the intervention,animal models were created.In the sham-operated group,the rats were only fixed on the operating board and the vessels were blunt-se pa rated.In the other groups,the right cerebral ischemia-repe rfusion injury model was established by the middle cerebral artery occlusion method and the Zea Longa suture method.Twenty-four hours after the modeling,the modified neurological severity scoring was performed to determine the damage.TUN EL staining method was then used to measure cell apoptosis in the rat cerebral cortex.Ultrastructure of the rat cerebral co rtex was observed under electron microscopy.RESULTS AND CONCLUSION:Compared with the sham-operated group,the modified neurological severity sco

关 键 词：脑缺血再灌注 细胞凋亡 microRNA-124 通督调神 NOTCH通路 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R255.2