长春西汀注射液在小鼠永久性脑缺血小胶质细胞表型转化中的作用研究  

Vinpocetine Injection Reduces Cerebral Ischemic Injury in Mice by Regulating Microglia Polarization

作　　者：陈青芳 赵顺英 温少红 董雯 刘文倩 龚婷 陈文涛 刘向荣 王拥军 CHEN Qing-Fang;ZHAO Shun-Ying;WEN Shao-Hong;DONG Wen;LIU Wen-Qian;GONG Ting;CHEN Wen-Tao;LIU Xiang-Rong;WANG Yong-Jun(China National Clinical Research Center for Neurological Diseases,Beijing 100070,China;Department of Neurology,Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China;Beijing Institute for Brain Disorders,Beijing 100070,China;Beijing Key Laboratory of Translational Medicine for Cerebrovascular Disease,Beijing 100070,China)

机构地区：[1]国家神经系统疾病临床医学研究中心,北京100070 [2]北京首都医科大学附属北京天坛医院神经病学中心 [3]北京脑重大疾病研究院卒中研究所 [4]北京市脑血管病转化医学重点实验室

出　　处：《中国卒中杂志》2021年第11期1156-1163,共8页Chinese Journal of Stroke

基　　金：国家自然科学基金项目(81901176,81871021)。

摘　　要：目的研究长春西汀注射液对小鼠永久性大脑中动脉远端缺血后神经功能恢复及小胶质细胞表型转化的影响。方法36只雄性C57BL/6J小鼠随机分为永久性脑缺血组6只、生理盐水组12只、长春西汀组12只和假手术组6只,前三组用高频电刀凝断小鼠右侧大脑中动脉远端,制作永久脑缺血模型,假手术组仅暴露大脑中动脉远端,不凝断血管。模型成功后,生理盐水组尾静脉注射生理盐水,每次150μL,每天1次,持续14 d;长春西汀组尾静脉注射长春西汀注射液,每次150μL(4.55 mg/kg),每天1次,持续14 d。模型成功后3 d、5 d、7 d、9 d、11 d和14 d进行改良加西亚评分和转棒测试评价小鼠感觉和运动神经功能;模型成功后14 d用免疫荧光标记神经元,评价各组神经元损伤情况,免疫荧光染色梗死周围小胶质细胞表型标志物Iba1、CD16/32和CD206的表达,评价M1型(Iba1及CD16/32阳性)和M2型(Iba1及CD206阳性)小胶质细胞表型转化情况。结果长春西汀组小鼠模型成功后11 d和14 d的改良加西亚评分和14 d的转棒测试中的时间及速度测试结果均优于永久性脑缺血组,差异均有统计学意义。长春西汀组14 d时神经元损伤较永久性脑缺血组(P=0.008)和生理盐水组(P=0.037)减轻。永久缺血组(P<0.001)和生理盐水组(P=0.005)M1型小胶质细胞表达高于假手术组;长春西汀组M1型小胶质细胞表达低于永久缺血组(P<0.001)和生理盐水组(P=0.038)。长春西汀组M2型小胶质细胞表达高于假手术组、永久性脑缺血组和生理盐水组(均P<0.001)。结论长春西汀注射液可能通过促进小胶质细胞表型由促炎向抗炎转变,减少神经元损伤,从而在小鼠永久性脑缺血后发挥神经保护和促进功能恢复的作用。Objective To study the effect of vinpocetine injection on the neurological function recovery and microglia polarization after permanent ischemia of distal middle cerebral artery(MCA)in mice.Methods Thirty six male C57BL/6J mice were randomly divided into permanent cerebral ischemia group(n=6),normal saline group(n=12),vinpocetine group(n=12)and sham group(n=6).The distal right MCA occlusion models were established by electrocoagulation in the first three groups.In the sham group,the distal of the MCA was exposed without electrocoagulation.Vinpocetine injection(4.55 mg/kg,150μL)or equal volume normal saline was injected via the caudal vein 20 minutes after ischemia for 5 minutes,and was administered continuously once a day for 14days.Modified Garcia scores and rotarod test were performed to evaluate the sensory and motor neurological function of mice at 3,7,9,11 and 14 days after the surgery.Neuron injury and Iba1 with CD16/32(M1 microglia,pro-inflammatory)or CD206(M2 microglia,anti-inflammatory)expression in cortex were evaluated by immunofluorescence staining at 14 days after ischemia.Results Vinpocetine injection improved modified Garcia scores at 11 and 14 days,quickened the running time and increased the speed of rotarod test at 14 days,compared with permanent cerebral ischemia group,with all statistical differences.Vinpocetine injection alleviate neuron injury compared to the permanent cerebral ischemia group(P=0.008)and normal saline treatment group(P=0.039).Vinpocetine injection decreased the expression of M1 microglia compared to the permanent cerebral ischemia group(P<0.001)and normal saline treatment group(P=0.038),and increased the expression of M2 microglia,compared to the other three groups(all P<0.001).Conclusions Vinpocetine injection may have protective effect against cerebral ischemia injury after permanent cerebral ischemia in mice by regulating microglia M1/M2 polarization from proinflammatory to anti-inflammatiory response.

关 键 词：脑缺血 长春西汀 大脑中动脉远端栓塞 小胶质细胞极化 小鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]