摘要
旨在探讨《古今录验》续命汤对急性脑梗死大鼠脑缺血性损伤及血管新生的影响。以SD大鼠为研究对象,随机分为6组:假手术组,模型组,续命汤低、中、高剂量组,丁苯酞组。大鼠大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型成功建立后,各组大鼠每天分别灌胃给予蒸馏水,续命汤5.13、10.26、20.52 g·kg~(-1)药液、丁苯酞混悬液(0.06 g·kg~(-1))。连续干预7 d后,进行末次神经功能评分,处死所有大鼠,采集脑组织样本。通过神经功能评分和2,3,5-氯化三苯基四氮染色评估脑缺血损伤程度;苏木素-伊红染色观察脑组织病理学形态改变;透射电子显微镜观察缺血半暗区皮层脑组织神经元和微血管内皮细胞(endothelial cells,ECs)超微结构;免疫荧光检测缺血半暗区皮层脑组织血管性血友病因子(von Willebrand factor,vWF)和造血祖细胞抗原CD34(hematopoietic progenitor cell antigen CD34,CD34)的蛋白表达情况;实时荧光定量PCR和免疫印迹法检测缺血半暗区皮层脑组织Runt相关转录因子1(Runt-related transcription factor 1,RUNX1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、血管生成素-1(angiopoietin-1,Ang-1)、血管生成素-2(angiopoietin-2,Ang-2)、VEGF受体2(VEGF receptor 2,VEGFR2)mRNA和蛋白的表达情况。结果表明,与假手术组相比,模型组大鼠神经功能评分、脑梗死面积显著升高(P<0.01);缺血半暗区皮层脑组织病理学形态结构、神经元与微血管ECs超微结构受损;RUNX1、VEGF、Ang-1、Ang-2、VEGFR2 mRNA表达均显著升高(P<0.01);vWF、CD34、RUNX1、VEGF、Ang-1、Ang-2、VEGFR2蛋白表达均显著升高(P<0.05或P<0.01)。与模型组相比,续命汤高剂量组、丁苯酞组神经功能缺损评分和脑梗死面积显著降低(P<0.01);缺血半暗区皮层脑组织病理学形态结构、神经元与微血管ECs超微结构损伤情况得到改善;RUNX1、VEGF、Ang-1、Ang-2、VEGFR2 mRNA表达显著升高(P<0.01);vW
This paper aims to explore the effect of Xuming Decoction in the Records of Proved Prescriptions,Ancient and Modern on cerebral ischemic injury and angiogenesis in the rat model of acute cerebral infarction.SD rats were randomized into 6 groups:sham group,model group,low-,medium-,and high-dose(5.13,10.26,and 20.52 g·kg-1,respectively)Xuming Decoction groups,and butylphthalide(0.06 g·kg-1)group.After the successful establishment of the rat model by middle cerebral artery occlusion(MCAO),rats in the sham and model groups were administrated with distilled water and those in other groups with corresponding drugs for 7 consecutive days.After the neurological function was scored,all the rats were sacrificed,and the brain tissue samples were collected.The degree of cerebral ischemic injury was assessed by the neurological deficit score and staining with 2,3,5-triphenyltetrazolium chloride.Hematoxylin-eosin staining was performed to observe the pathological changes in the brain.Transmission electron microscopy was employed to observe the ultrastructures of neurons and microvascular endothelial cells(ECs)on the ischemic side of the brain tissue.Immunofluorescence assay was employed to detect the expression of von Willebrand factor(vWF)and hematopoietic progenitor cell antigen CD34(CD34)in the ischemic brain tissue.Real-time PCR and Western blot were employed to determine the mRNA and protein levels,respectively,of Runt-related transcription factor 1(RUNX1),vascular endothelial growth factor(VEGF),angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),and VEGF receptor 2(VEGFR2)in the ischemic brain tissue.The results showed that compared with the sham group,the model group showed increased neurological deficit score and cerebral infarction area(P<0.01),pathological changes,and damaged ultrastructure of neurons and microvascular ECs in the ischemic brain tissue.Furthermore,the modeling up-regulated the mRNA levels of RUNX1,VEGF,Ang-1,Ang-2,and VEGFR2(P<0.01)and the protein levels of vWF,CD34,RUNX1,VEGF,Ang-1,Ang-2,and VEGFR2(P<0.05
作者
罗政
李定祥
蔡昱哲
刘艺璇
阳晶晶
吴琴
张亚男
陈静
邓奕辉
LUO Zheng;LI Ding-xiang;CAI Yu-zhe;LIU Yi-xuan;YANG Jing-jing;WU Qin;ZHANG Ya-nan;CHEN Jing;DENG Yi-hui(School of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases,Hunan University of Chinese Medicine,Changsha 410208,China;School of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第8期2178-2187,共10页
China Journal of Chinese Materia Medica
基金
湖南省科技创新团队项目(2020RC4050)
湖南省教育厅科学研究项目(21A0228)
湖南省中医药科研计划项目(E2022010)。
关键词
急性脑梗死
《古今录验》续命汤
血管新生
RUNX1
VEGF
acute cerebral infarction
Xuming Decoction in the Records of Proved Prescriptions,Ancient and Modern
angiogenesis
RUNX1
VEGF
