中医康复2024，Vol.1Issue(2)：8-13,6.DOI:10.19787/j.issn.2097-3128.2024.02.002
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 of Nestin and GFAP Proteins in Rats with Cerebral Ischemia/Reperfusion Model
摘要
Abstract
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 infarction was calculated by TTC staining method;Western Blot were used to detect the expression of Nestin,GFAP in the infarcted brain tissue and CD63 in the serum.Results:After 7d of intervention,the sham operation group did not show symptoms of neurological deficits,the neurological deficits score of the model group was significantly higher(P＜0.01),the neurological deficits score of the"Buqihuoxue"compounds group was significantly lower than that of the model group(P＜0.05),and the score was significantly higher after the application of GW4869(P＜0.01);TTC staining showed that the brain tissue in the sham operation group was normal red,and no pale infarct foci were seen,while pale infarct foci were seen on the right side of the brain in the model group,"Buqihuoxue"compounds group and"Buqihuoxue"compounds+exosome inhibitor group;the infarct volume of the model group,the"Buqihuoxue"compounds group and"Buqihuoxue"compounds+exosome inhibitor group was significantly increased compared with that of the sham operation group(P＜0.01);and the volume in"Buqihuoxue"compounds group was significantly lower than that in the model group(P＜0.05).Compared with the"Buqihuoxue"compounds group,the volume in the"Buqihuoxue"compounds+exosome inhibitor group was sig-nificantly higher(P＜0.01);HE showed that the sham operation group was uniformly stained,tissue intacted,and the neuronal cells were neatly ar-ranged with conical shape,the nucleus was large in the middle with a pronounced nucleoli.In the model group,the brain nerve cells irregularly ar-ranged,neurons obviously swollen,the nuclei consolidation.Vacuole-like changes of different size existed in the cell space,combined with intersti-tial edema.Compared with the model group,neuronal necrosis reduction and tissue edema were milder in the"Buqihuoxue"compounds and"Buqi-huoxue"compounds+exosome inhibitor group.Compared with the"Buqihuoxue"compounds+exosome inhibitor group,the neuronal necrosis reduc-tion and tissue edema was milder in the"Buqihuoxue"compounds group;Western Blot showed that the expression of GFAP and CD63 was signifi-cantly lower in the model group compared with the sham operation group(P＜0.05),while the expression in"Buqihuoxue"compounds group were si-ginificantly improved(P＜0.05).However,the expression of GFAP and CD63 in"Buqihuoxue"compounds group were decreased after the addition of GW4869.The expression of Nestin was just intended to be similar to GFAP and CD63,but no significant difference was found(P＞0.05).Conclusion:"Buqihuoxue"compounds can reduce the volume of cerebral infarction,increase the expression of GFAP proteins in cerebral infarcted tissues,and improve the degree of neurological deficits after ischemia/reperfusion in rats,which may be related to the promotion of exosome secretion.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/CD63Key words
"Buqihuoxue"compounds/ischemia-reperfusion/neurologic deficits/exosomes/Nestin/GFAP/CD63分类
医药卫生引用本文复制引用
刘通,廖翔宇,陈玉宁,蒋颖,陈琼君,熊亮,邝伟川,文希,刘悦..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[J].中医康复,2024,1(2):8-13,6.基金项目
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 （）
