补气活血合剂干预脑缺血再灌注模型大鼠相关因子及自噬蛋白的表达  

Buqi Huoxue Compounds intervene with the expression of related factors and autophagy related proteins in a rat model of cerebral ischemia/reperfusion

作　　者：陈玉宁 蒋颖 廖翔宇 陈琼君 熊亮 刘悦 刘通[1,2] Chen Yuning;Jiang Ying;Liao Xiangyu;Chen Qiongjun;Xiong Liang;Liu Yue;Liu Tong(The Fifth Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510095,Guangdong Province,China;Department of Acupuncture and Rehabilitation,Guangdong Second Hospital of Traditional Chinese Medicine,Guangzhou 510095,Guangdong Province,China)

机构地区：[1]广州中医药大学第五临床医学院,广东省广州市510095 [2]广东省第二中医院针灸康复科,广东省广州市510095

出　　处：《中国组织工程研究》2025年第6期1152-1158,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省中医药局专项研究项目(20203001),项目负责人:刘悦;广东省自然科学基金面上项目(2022A1515011676),项目负责人:刘悦;广东省自然科学基金省企联合基金(2022A1515220012),项目负责人:刘通;国家自然科学基金面上项目(82174482),项目负责人:刘通;中央财政转移支付地方项目(602023057),项目参与人:刘悦,刘通;2024广东省刘悦名中医工作室建设项目(粤中医办函[2023]108号),项目负责人:刘悦。

摘　　要：背景:补气活血合剂治疗气虚痰瘀型缺血性脑卒中有显著的临床疗效,然而其具体起效机制尚不明确。目的:观察补气活血合剂对脑缺血再灌注模型大鼠血管内皮细胞生长因子、碱性成纤维细胞生长因子、脑源性神经营养因子及自噬蛋白Beclin1、p62表达的影响。方法:取40只雄性SD大鼠,采用随机数字表法分为假手术组、模型组、补气活血合剂组与自噬抑制剂组,每组10只。模型组、补气活血合剂组与自噬抑制剂组建立大脑缺血再灌注损伤模型,补气活血合剂组大鼠再灌注2 h后灌胃给予补气活血合剂(6.49 g/kg,每天分3次给药),自噬抑制剂组大鼠再灌注2 h后灌胃给予补气活血合剂(6.49 g/kg,每天分3次给药)+腹腔注射自噬抑制剂3-甲基腺嘌呤(灌胃前2 h注射,灌胃第1-3天注射),假手术组、模型组灌胃给予等量生理盐水,连续灌胃给药7 d。末次给药24 h后,进行大鼠神经功能、脑梗死体积、脑组织形态及大脑缺血皮质区血管内皮细胞生长因子、碱性成纤维细胞生长因子、脑源性神经营养因子及自噬蛋白Beclin1、p62表达检测。结果与结论:①Zea-Longa评分结果显示,造模后大鼠神经功能受到严重损伤,补气活血合剂组大鼠神经功能损伤轻于模型组、自噬抑制剂组(P<0.05);②TTC染色结果显示,模型组、补气活血合剂组与自噬抑制剂组大鼠均可见大脑梗死灶,补气活血合剂组大鼠脑梗死体积低于模型组、自噬抑制剂组(P<0.05);③缺血侧脑组织苏木精-伊红染色结果显示,模型组神经细胞间有较大的空隙且排列不整齐,神经细胞出现胞质凝集、核固缩现象;补气活血合剂组和自噬抑制剂组神经细胞间隙减小且排列较规律,存在部分细胞出现胞质凝集、核固缩表现;④免疫组化染色结果显示,血管内皮生长因子多表达在神经元细胞、神经胶质细胞及毛细血管内皮;碱性成纤维细胞生长因子和脑源性BACKGROUND:Buqi Huoxue Compounds have significant clinical efficacy in treating ischemic stroke with Qi deficiency and phlegm stasis;however,the exact mechanism of action is not clear.OBJECTIVE:To observe the effect of Buqi Huoxue Compounds on the expression of vascular endothelial growth factor,basic fibroblast growth factor,brainderived neurotrophic factor and autophagy related protein Beclin1 and p62 in a rat model of cerebral ischemia/reperfusion.METHODS:Forty male Sprague-Dawley rats were randomly divided into sham operation group,model group,Buqi Huoxue Compounds group and autophagy inhibitor group,with 10 rats in each group.In the latter three groups,a rat model of cerebral ischemia/reperfusion injury was established.The Buqi Huoxue Compounds group was intragastrically given Buqi Huoxue Compounds(6.49 g/kg,administered three times a day)2 hours after reperfusion;the autophagy inhibitor group was intragastrically given Buqi Huoxue Compounds(6.49 g/kg,administered three times a day)2 hours after reperfusion and intraperitoneally given 3-methyladenine 2 hours before gavage and at days 1-3 of gavage.The sham operation group and model group were given equal amounts of saline by gavage for 7 consecutive days.Neurological function,cerebral infarct volume,brain tissue morphology and expression of vascular endothelial growth factor,basic fibroblast growth factor,brain-derived neurotrophic factor and autophagy-related proteins Beclin1 and p62 in the ischemic cortical region of rats were detected at 24 hours after the final administration.RESULTS AND CONCLUSION:Zea-Longa scoring results showed that the neurological function of rats was severely damaged after modeling and neurological deficit of rats in the Buqi Huoxue Compounds group was less than that in the model group and the autophagy inhibitor group(P<0.05).TTC staining showed that cerebral infarct foci were observed in the model group,Buqi Huoxue Compounds group,and autophagy inhibitor group,and the cerebral infarct volume in the Buqi Huoxue Compounds group was

关 键 词：补气活血合剂 缺血再灌注 神经功能缺损 自噬 血管内皮生长因子 碱性成纤维细胞生长因子 脑源性神经营养因子 BECLIN1 P62 

分 类 号：R453.9[医药卫生—治疗学] R363[医药卫生—临床医学] R741.02