红景天苷对脑缺血再灌注模型大鼠内皮细胞屏障的保护作用及机制研究  

Protection of salidroside on endothelial cell barrier in cerebral ischemia-reperfusion model rats

作　　者：宋文婷[1] 曹慧 张业昊 郑晓宇 刘建勋[1] SONG Wen-ting;CAO Hui;ZHANG Ye-hao;ZHENG Xiao-yu;LIU Jian-xun(Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,北京市中药药理重点实验室,北京100091 [2]北京中医药大学,北京100029

出　　处：《中国中药杂志》2022年第19期5284-5291,共8页China Journal of Chinese Materia Medica

基　　金：中国中医科学院科技创新工程项目(CI2021A04614)。

摘　　要：该文旨在观察红景天苷对脑缺血再灌注大鼠的治疗作用,探究红景天苷对脑缺血再灌注模型大鼠内皮细胞屏障的保护作用及其机制.实验中将SD大鼠随机分为假手术组,模型组,红景天苷高、中、低剂量(10、5、2.5 mg·kg^(-1))组,应用线栓法建立大鼠脑缺血再灌注模型,使用脑梗死面积、神经行为学评价、脑含水量作为药效指标.机制探讨实验部分使用红景天苷高、低剂量组为给药组.使用HE染色观察病理学形态,透射电镜观察血管内皮细胞超微结构;检测血清中一氧化氮(nitric oxide,NO)含量、凝血四项值及血浆中血管性血友病因子(von willebrand factor,vWF)含量;再利用免疫印迹(Western blot,WB)、免疫荧光(immunofluorescence,IF)检测大鼠皮层组织中紧密连接蛋白(ZO-1、occludin、claudin-1)和MMP-9表达.结果显示,与假手术组比较,模型组大鼠出现明显的神经功能缺损,右侧脑组织出现明显梗死灶,脑含水量明显升高;与模型组比较,红景天苷高、低剂量组大鼠的神经行为学评分有不同程度的降低,红景天苷高、中、低剂量组大鼠脑梗死面积明显减小,大鼠脑组织含水量也明显降低.HE染色及透射电镜结果显示,与假手术组比较,大鼠脑缺血再灌注后,皮层组织中出现神经元坏死,内皮细胞原有生理结构被破坏,内皮细胞间紧密连接被瓦解;与模型组比较,红景天苷高、低剂量组皮层神经元损伤减轻,内皮细胞生理结构相对完整.与假手术组比较,模型组大鼠血清中NO水平显著降低且血浆中vWF、纤维蛋白原(fibrino-gen,FIB)表达水平也明显升高,凝血酶时间(thrombin time,TT)、凝血酶原时间(prothrombin time,PT)明显缩短.与模型组比较,红景天苷高、低剂量组血清中NO含量升高,血浆中FIB、vWF水平降低,TT、PT也明显延长.WB、IF结果显示,与假手术组比较,模型组大鼠皮层脑组织中内皮细胞间的ZO-1、occludin、claudin-1表达水平显This study aims to observe the therapeutic effect of salidroside on cerebral ischemia-reperfusion(I/R) model rats, and to specifically explore the protection of salidroside on endothelial cell barrier after I/R and the mechanism. In the experiment, SD rats were randomized into sham group, model group, and high-, medium-, and low-dose(10, 5, and 2.5 mg·kg) salidroside groups. The suture method was used to induce I/R in rats. The infarct area, neurobehavioral evaluation, and brain water content were used to evaluate the efficacy of salidroside. As for the experiment on the mechanism, high-dose and low-dose salidroside groups were designed. The pathological morphology was observed based on hematoxylin and eosin(HE) staining, and ultrastructure of vascular endothelial cells based on transmission electron microscopy. The content of nitric oxide(NO) in serum, four indexes of blood coagulation, and the content of von Willebrand factor(vWF) in plasma were measured. Western blot(WB) and immunofluorescence(IF) were employed to determine the expression of tight junction proteins(ZO-1, occluding, and claudin-1) and matrix metalloproteinase 9(MMP-9) in the cortex. The results showed that the model group had obvious neurological deficit, obvious infarct in the right brain tissue, and significant increase in water content in brain tissue compared with the sham group. Compared with the model group, high-dose and low-dose salidroside groups showed decrease in neurobehavioral score, and the high-, medium-, and low-dose salidroside groups demonstrated obviously small infarct area and significant decrease in water content in brain tissue. The results of HE staining and transmission electron microscopy showed that rats had necrosis of neurons, damage of original physiological structure of endothelial cells, and disintegration of the tight junction between endothelial cells after I/R compared with the sham group. Compared with the model group, the high-dose and low-dose salidroside groups showed alleviation of neuron injury and intac

关 键 词：红景天苷 脑缺血再灌注 内皮细胞 内皮屏障 

分 类 号：R285.5[医药卫生—中药学]