补气活血合剂对脑缺血再灌注模型大鼠血清外泌体及Nestin、GFAP蛋白表达的影响  

Effects of "Buqihuoxue" Compounds on Serum Exosomes and the Expression ofNestin and GFAP Proteins in Rats with Cerebral Ischemia/Reperfusion Model

作　　者：刘通 廖翔宇 陈玉宁 蒋颖 陈琼君 熊亮 邝伟川 文希[1,2] 刘悦 LIU Tong;LIAO Xiang-yu;CHEN Yu-ning;JIANG Ying;CHEN Qiong-jun;XIONG Liang;KUANG Wei-chuan;WEN Xi;LIU Yue(The Fifth Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou,Guangdong 510095;Guangdong Second Traditional Chinese Medicine Hospital(Guangdong Research Institute of Traditional Chinese Medicine Manuracturing Tech‐nology),Guangzhou,Guangdong 510095)

机构地区：[1]广州中医药大学第五临床医学院,广东广州510095 [2]广东省第二中医院(广东省中医药工程技术研究院),广东广州510095

出　　处：《中医康复》2024年第2期8-13,共6页Traditional Chinese Medicine Rehabilitation

基　　金：广东省中医药局专项研究项目,编号:20203001;国家自然科学基金,编号:82174482;广东省自然科学基金,编号:2022A1515011676。

摘　　要：目的:观察补气活血合剂对脑缺血再灌注模型大鼠血清外泌体及巢蛋白(Nestin)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)表达的影响。方法:将40只雄性SD大鼠随机分为假手术组、模型组、补气活血合剂组及补气活血合剂+外泌体抑制剂GW4869组(后简称合剂+GW4869组)各10只,模型组、补气活血合剂组及合剂+GW4869组均采用大脑中动脉线栓法复制大鼠大脑中动脉梗塞模型,假手术组仅给予颈内动脉、颈外动脉及颈总动脉的解剖分离而不进行线栓插入,缺血2h后拔除线栓进行再灌注;补气活血合剂及合剂+GW4869组分别在再灌注2h后开始灌胃补气活血合剂(10mL/kg,3次/日),合剂+GW4869组在每次灌胃前给予外泌体抑制剂GW4869腹腔注射(2.5mg/kg·d),假手术组及模型组给予灌胃等量生理盐水,四组均连续干预7天。7天后,分别记录各组大鼠干预前后的神经功能缺损评分,TTC染色法计算干预后各组大鼠脑梗死体积,HE染色法评估各组大鼠神经组织形态学变化,Western Blot法检测大鼠梗死区组织Nestin、GFAP蛋白及血清外泌体CD63蛋白的表达水平。结果:干预7天后,假手术组未表现出神经功能缺损症状,模型组神经功能缺损评分显著升高(P<0.01),补气活血合剂组神经功能缺损评分较模型组显著降低(P<0.05),应用GW4869后,神经功能缺损评分较补气活血合剂组显著升高(P<0.01)。TTC染色结果显示:假手术组大鼠脑组织成正常红色,未见肉眼苍白梗死灶,模型组、补气活血合剂组、合剂+GW4869组均可见右侧大脑苍白梗死灶;与假手术组相比,模型组、补气活血合剂组、合剂+GW4869组大鼠的脑梗死体积比均显著增加(P<0.01);与模型组相比,补气活血合剂组的大鼠脑梗死体积比显著降低(P<0.05);与补气活血合剂组相比,合剂+GW4869组大鼠的脑梗死体积比均显著增加(P<0.01)。HE结果显示:假手术组染色均匀,组织完整,神经细胞形Objective:To observe the effect of"Buqihuoxue"compounds on serum exosomes and the expression of Nestin and GFAP Proteins in rats with cerebral ischemia/reperfusion model.Methods:40 male SD rats were randomly divided into the sham operation group,model group,"Buqi‐huoxue"compounds group and"Buqihuoxue"compounds+exosome inhibitor group,with 10 rats in each group.In the model group,the"Buqi‐huoxue"compounds group and the"Buqihuoxue"compounds+exosome inhibitor group,the rat model of middle cerebral artery occlusion was made by suturing of the middle cerebral artery,while rats in the sham operation group was just given the dissection and isolation of the internal carotid ar‐tery,external carotid artery and common carotid artery.After 2 hours of cerebral ischemia,the middle artery suture was removed for reperfusion.Rats in"Buqihuoxue"compounds group and"Buqihuoxue"compounds+exosome inhibitor group were given"Buqihuoxue"compounds after 2 hours of reperfusion(10mL/kg,3 times/day),the exosome inhibitor GW4869 was given intraperitoneally before each gavage in the"Buqihuoxue"com‐pounds+exosome inhibitor group(2.5mg/kg·d).The sham operation group and the model group were given an equal amount of saline by gavage,and all four groups were intervened for 7 consecutive days.The neurological deficit scores of the rats in each group were recorded before and after the in‐tervention, respectively;HE staining method was used to assess the morphology changes of the neurological tissues;the volume of cerebral infarctionwas calculated by TTC staining method;Western Blot were used to detect the expression of Nestin, GFAP in the infarcted brain tissue and CD63 inthe serum. Results: After 7d of intervention, the sham operation group did not show symptoms of neurological deficits, the neurological deficits scoreof the model group was significantly higher (P<0.01), the neurological deficits score of the "Buqihuoxue" compounds group was significantly lowerthan that of the model group (P<0.05), and the score was significantly higher

关 键 词：补气活血合剂 缺血再灌注 神经功能缺损 外泌体 NESTIN GFAP CD63 

分 类 号：R285.5[医药卫生—中药学]