摘要
该研究旨在探究脑心通胶囊是否通过促进血管新生改善多发性脑梗死损伤及并发心肌损伤,发挥脑心同治作用。将SD雄性大鼠随机分为6组:假手术组,模型组,脑心通胶囊高、中、低剂量(440、220、110 mg·kg^(-1))组及尼莫地平(10.8 mg·kg^(-1))组,通过注射自体血栓块制备多发性脑梗死大鼠模型,造模7 d后进行取材和指标检测,包括多发性脑梗死大鼠模型评价,神经功能评分、抓握力测试、转棒测试检测神经运动功能,苏木素-伊红(HE)染色、尼氏染色、Masson染色观察脑、心肌组织形态结构,网络药理学筛选脑心通胶囊改善多发性脑梗死和心肌损伤机制,透射电镜观察脑组织神经元、心肌组织心肌细胞超微结构,原位末端标记法(TUNEL)染色检测脑神经元细胞凋亡率,检测心肌细胞ROS含量,免疫荧光检测脑组织和心肌组织血小板-内皮细胞黏附分子(CD31)和细胞核增殖抗原67(Ki67)、造血干细胞抗原(CD34)、缺氧诱导因子-1α(HIF-1α)表达,蛋白免疫印迹(WB)法、实时荧光定量逆转录聚合酶链反应(RT-qPCR)检测HIF-1α、血管内皮生长因子(VEGF)、血管内皮细胞生长因子受体2(VEGFR2)、酪氨酸蛋白激酶(Src)、碱性成纤维细胞生长因子(bFGF)、血管生成素-1(Ang-1)、血管生成素受体酪氨酸激酶-2(Tie-2)表达。相比较模型组而言,脑心通胶囊中剂量组神经功能评分明显降低,抓握力增大和在棒时间显著延长,脑、心肌组织病理染色损伤减轻;神经元和心肌细胞内线粒体增多,形态和排列较为正常;脑神经元细胞凋亡减轻,心肌细胞ROS含量下降;脑组织和心脏的微血管密度增加,新生血管内皮细胞增多,CD31和Ki67的表达重叠区域增多;脑组织和心肌组织的HIF-1α、VEGF、VEGFR2、Src、Ang-1、Tie-2和bFGF蛋白及mRNA相对表达量均升高。脑心通胶囊可能通过介导HIF-1α/VEGF表达促进血管新生改善多发性脑梗死损伤及并发心肌损伤。
This study aims to explore whether Naoxintong Capsules improve multiple cerebral infarctions and myocardial injury via promoting angiogenesis,thereby exerting a simultaneous treatment effect on both the brain and heart.Male SD rats were randomly divided into six groups:sham-operated group,model group,high-dose,medium-dose,and low-dose groups of Naoxintong Capsules(440,220,and 110 mg·kg^(-1)),and nimodipine group(10.8 mg·kg^(-1)).Rat models of multiple cerebral infarctions were established by injecting autologous thrombus,and samples were collected and tested seven days after modeling.Evaluations included multiple cerebral infarction model assessments,neurological function scores,grip strength tests,and rotarod tests,so as to evaluate neuromotor functions.Morphological structures of brain and heart tissue were observed using hematoxylin-eosin(HE)staining,Nissl staining,and Masson staining.Network pharmacology was employed to screen the mechanisms of Naoxintong Capsules in improving multiple cerebral infarctions and myocardial injury.Neuronal and myocardial cell ultrastructures were observed using transmission electron microscopy.Apoptosis rate in brain neuronal cells was detected by TdT-mediated dUTP nick end labeling(TUNEL)staining,and reactive oxygen species(ROS)levels in myocardial cells were measured.Immunofluorescence was used to detect the expression of platelet endothelial cell adhesion molecule-1(CD31),antigen identified by monoclonal antibody Ki67(Ki67),hematopoietic progenitor cell antigen CD34(CD34),and hypoxia inducible factor-1α(HIF-1α)in brain and myocardial tissue.Western blot,and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the expression of HIF-1α,vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 2(VEGFR2),sarcoma(Src),basic fibroblast growth factor(bFGF),angiopoietin-1(Ang-1),and TEK receptor tyrosine kinase(Tie-2).Compared with the model group,the medium-dose group of Naoxintong Capsules showed significantly lower neurological function scores,increased grip strength,and prolonged time on the rotarod.Pathological damage in brain and heart tissue was reduced,with increased and more orderly arranged mitochondria in neurons and cardiomyocytes.Apoptosis in brain neuronal cells was decreased,and ROS levels in cardiomyocytes were reduced.The microvascular density and endothelial cells of new blood vessels in brain and heart tissue increased,with increased overlapping regions of CD31 and Ki67 expression.The relative protein and mRNA expression levels of HIF-1α,VEGF,VEGFR2,Src,Ang-1,Tie-2,and bFGF were elevated in brain tissue and myocardial tissue.Naoxintong Capsules may improve multiple cerebral infarctions and myocardial injury by mediating HIF-1α/VEGF expression to promote angiogenesis.
作者
张晓璐
尚津锋
文胤琏
黄贵金凤
王伯洪
韦婉婷
陈文彬
刘欣
ZHANG Xiao-lu;SHANG Jin-feng;WEN Yin-lian;HUANG Gui-jin-feng;WANG Bo-hong;WEI Wan-ting;CHEN Wen-bin;LIU Xin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Bejing 102488,China)
出处
《中国中药杂志》
北大核心
2025年第7期1889-1899,共11页
China Journal of Chinese Materia Medica
基金
北京中医药大学基本科研业务费项目(揭榜挂帅项目)(2022-JYB-JBZR-005)。
关键词
多发性脑梗死
脑心通胶囊
脑心同治
血管生成
低氧诱导因子-1Α
multiple cerebral infarction
Naoxintong Capsules
simultaneous treatment of heart and brain
angiogenesis
hypoxiainducible factor-1α
