[1]乔建新,刘熙鹏,刘　明,等.加贝酯对脑缺血再灌注大鼠脑组织病理学改变的影响[J].新乡医学院学报,2021,38(3):226-232.[doi:10.7683/xxyxyxb.2021.03.005]
　QIAO Jianxin,LIU Xipeng,LIU Ming,et al.Effect of gabexate mesilate on pathological changes of brain tissue of rats with cerebral ischemia reperfusion[J].Journal of Xinxiang Medical University,2021,38(3):226-232.[doi:10.7683/xxyxyxb.2021.03.005]
点击复制

加贝酯对脑缺血再灌注大鼠脑组织病理学改变的影响
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年3
页码:
226-232
栏目:
基础研究
出版日期:
2021-03-05

文章信息/Info

Title:
Effect of gabexate mesilate on pathological changes of brain tissue of rats with cerebral ischemia reperfusion
作者:
乔建新; 刘熙鹏; 刘　明; 曹　兵
（河北北方学院附属第一医院神经外科，河北　张家口　075000）
Author(s):
QIAO Jianxin; LIU Xipeng; LIU Ming; CAO Bing
(Department of Neurosurgery，the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province，China)
关键词:
加贝酯; 脑缺血再灌注; 血脑屏障; 氧化应激; 细胞凋亡
Keywords:
gabexate mesilatecerebral ischemia-reperfusionblood-brain barrieroxidative stressapoptosis
分类号:
R289.11
DOI:
10.7683/xxyxyxb.2021.03.005
文献标志码:
A
摘要:
目的　研究加贝酯（GM）对脑缺血再灌注（IR）大鼠脑组织病理学改变的影响及其机制。方法　将240只成年雄性Sprague Dawley大鼠随机分为假手术组、IR组、尼莫地平（NMP）组和5、10、20 mg·kg-1·d-1 GM组，每组40只。除假手术组外，其余各组大鼠采用线栓法制备脑IR模型；假手术组大鼠除不在颈外动脉切口和插入线栓外，其余操作同模型组。造模完成后，5、10、20 mg·kg-1·d-1 GM组大鼠分别经尾静脉注射5、10、20 g·L-1的GM溶液，NMP组大鼠经尾静脉注射2 g·L-1的NMP溶液，假手术组和IR组大鼠经尾静脉注射生理盐水，各组注射剂量均为1 mL·kg-1，每日1次，连续7 d。末次给药2 h后，采用Zausinger 评分法评估各组大鼠神经功能，伊文思蓝渗透法测定各组大鼠血脑屏障（BBB）通透性，干湿比重法测定各组大鼠脑组织含水量，苏木精伊红染色法、脱氧核糖核苷酸末端转移酶介导的缺口末端标记法观察各组大鼠脑组织病理学变化和细胞凋亡情况；透射电子显微镜观察各组大鼠神经细胞超微结构变化；生物化学分析法测定各组大鼠脑组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性和丙二醛（MDA）水平；Western blot法检测各组大鼠脑组织中核因子-κB（NF-κB）蛋白相对表达量。结果　假手术组大鼠脑组织形态结构及神经细胞超微结构均正常；IR组大鼠脑组织可见神经细胞数量减少、间隙增大、排列无序，胞体皱缩呈空泡状变性、细胞核固缩深染，细胞膜破裂、细胞质溶解，核仁边界不清，线粒体肿胀、嵴断裂溶解，粗面内质网扩张、颗粒脱落等；与IR组比较，各干预组大鼠脑组织形态结构病变、细胞损伤及神经细胞超微结构病变均明显改善。与假手术组比较，IR组大鼠神经功能缺损评分降低，BBB通透性和脑组织含水量升高，神经细胞凋亡指数升高，脑组织中SOD、GSH-Px活性降低，MDA水平升高，NF-κB蛋白相对表达量上调（P<0.05）。与ir组比较，5、10、20 mg·kg-1·d-1 GM组大鼠神经功能评分升高，BBB通透性和脑组织含水量下降，神经细胞凋亡指数降低，脑组织中SOD、GSH-Px活性升高，MDA水平降低，NF-κB蛋白相对表达量下调，且均呈剂量依赖性趋势。20 mg·kg-1·d-1 GM组大鼠除脑组织含水量外以上各指标的变化均优于NMP组。结论　GM对脑IR大鼠脑组织病理学改变、神经细胞超微结构病变具有抑制作用，对BBB通透性具有保护作用，其机制可能与降低氧化应激、下调NF-κB蛋白表达以及抑制细胞凋亡有关；较高剂量GM（20 mg·kg-1·d-1 ）对脑IR大鼠脑组织的保护作用优于NMP。undefined/html>
Abstract:
Objective　To study the effect of gabexate mesilate (GM) on pathological changes of brain tissue of the rats with cerebral ischemia-reperfusion (IR) and explore its possible mechanism.Methods　　Two hundred and forty adult male Sprague Dawley rats were randomly divided into sham operation group,IR group,nimodipine (NMP) group and 5,10,20 mg·kg-1·d-1 GM group,with 40 rats in each group.Except for the rats in the sham operation group,the rats in the other groups were made the brain IR model by the suture method;the rats in the sham operation group did not make the external carotid artery incision and insert the suture,the other operations were same as the model group.After modeling,the rats in the 5,10,20 mg·kg-1·d-1 GM groups were injected with 5,10,20 g·L-1 GM solution via the tail vein respectively;the rats in the NMP group were injected with 2 g·L-1 NMP solution via the tail vein;the rats in sham operation group and IR group were injected with normal saline via the tail vein;the injection dose in each group was 1 mL·kg-1,once a day for 7 days.Two hours after the last administration,the neurological function of rats in each group was evaluated by Zausinger score method;the permeability of blood brain barrier (BBB) of rats in each group was measured by Evans blue permeation method and the water content of brain tissue was measured by dry wet specific gravity method;the pathological change of brain tissue of rats in each group was observed by hematoxylin eosin staining and the apoptosis was observed by terminal-deoxynucleotidyl transferase mediated nick end labeling method;the ultrastructure of nerve cells in brain tissues of rats in each group were observed by transmission electron microscope;the superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA) content in brain tissue of rats in each group were determined by biochemical analysis;the relative expression level of nuclear factor-κB (NF-κB) protein in brain tissue of rats in each group was determined by Western blot.Results　In the sham operation group,the morphology of brain tissue and ultrastructure of nerve cells were normal.In the IR group,the number of nerve cells decreased,the gap of nerve cells increased,the arrangement of nerve cells was disordered;the cell body shrank and showed vacuolar degeneration,and the nucleus pyknosis and deep stained;the cell membrane of nerve cell ruptured,the cytoplasmic was dissolved;and the unclear nucleolus boundary,mitochondrial swelling,mitochondrial crista fracture and dissolution,rough endoplasmic reticulum expansion,rough endoplasmic reticulum granules desquamation could be seen in the nerve cells.Compared with the IR group,the morphological changes of brain tissue,cell damage and ultrastructural changes of nerve cells in each intervention group were significantly improved.Compared with the sham operation group,the neurological deficit scores of rats were lower;the BBB permeability,the water content in brain tissues,apoptosis index were higher;the SOD,GSH-Px activities in brain tissue were lower,and the MDA content in brain tissue was higher;the expression of NF-κB protein was increased in the IR group(P<0.05).Compared with IR group,the neurological function scores of rats increased,BBB permeability and water content of brain tissue decreased,apoptosis index of nerve cells decreased,SOD and GSH-Px activities in brain tissue increased,MDA content decreased,and the expression of NF-κB protein in brain tissue decreased in a dose-dependent manner in the 5,10,20 mg·kg-1·d-1 GM groups.The changes of the above indexes except for the brain water content in the 20 mg·kg-1·d-1 GM group were better than those in the NMP group.Conclusion　GM can inhibit the brain histopathological changes,ultrastructural changes of nerve cells and protect the BBB permeability of the IR rat.The mechanism may be related to the reduction of oxidative stress,down-regulating of NF-κB protein expression,and the inhibition of apoptosis.The protective effect of higher dose of GM (20 mg·kg-1·d-1)on the brain tissue of IR rats is better than that of NMP.

参考文献/References:

［1］　PETROVIC-DJERGOVIC D,GOONEWARDENA S N,PINSKY D.Inflammatory disequilibrium in stroke ［J］.Circ Res,2016,119(1):142-158.
［2］　HUANG L,CHEN C,ZHANG X,et al.Neuroprotective effect of curcumin against cerebral ischemia-reperfusion via mediating autophagy and inflammation［J］.J Mol Neurosci,2018,64(1):129-139.
［3］　谭诚,韩亮,李朝晖,等.毛蕊花糖苷对脑缺血再灌注损伤大鼠的治疗作用［J］.中华实验外科杂志,2019,36(8):1368-1370.
［4］　张楠,祁晓媛,潘思培,等.香叶木素对脑缺血再灌注损伤大鼠的保护作用研究［J］.中国免疫学杂志,2019,35(24):2996-3000,3007.
［5］　刘景诗,徐军美,刘永平,等.NF-κB在加贝酯对大鼠肝缺血再灌注损伤过程中的活化及意义［J］.中国现代医学杂志,2010,20(6):821-823.
［6］　王健,刘振芬.甲磺酸加贝酯在小肠缺血-再灌注损伤中的预防作用［J］.中国药业,2013,22(15):33-34.
［7］　潘洲佳,顾佳楠,谢竹青,等.脑缺血再灌注模型制作方法的研究进展［J］.医学综述,2019,25(14):2801-2805.
［8］　ZAUSINGER S,WESTERAIER T,PLESNILA N,et al.Neuroprotection in trasient focal cerebral ischemia by combination drug therapy andmild hypothermia:comparison with customary therapeutic regimen［J］.Stroke,2003,34(6):1526-1532.
［9］　邱珂,邓淑文,刘慧,等.丁苯酞对SD大鼠脑出血后血脑屏障的影响及其机制［J］.中南大学学报(医学版),2019,44(12):1321-1328.
［10］　马历历,李浩,孙立明,等.黄芪总黄酮对脑缺血再灌注损伤大鼠氧化应激、炎症、凋亡的影响［J］.中成药,2019,41(8):1811-1815.
［11］　任俊伟,杨琴.白藜芦醇对脑缺血/再灌注氧化应激损伤保护机制的研究进展［J］.中国药理学通报,2011,27(4):448-451.
［12］　刘慧芳,姜敏,马红霞.哮喘大鼠肺组织线粒体功能变化及其对氧化应激水平的影响［J］.新乡医学院学报,2018,35(8):662-665.
［13］　ZHENG J Q,ZHANG G R,LI J,et al.Neutrophil elastase inhibitor suppresses oxidative stress in obese asthmatic rats by activating Keap1/Nrf2 signaling pathway［J］.Eur Rev Med Pharmacol Sci,2019,23(1):361-369.
［14］　KHAN A M,KORZENIOWSKA B,GORSHKOV V,et al.Silver nanoparticle-induced expression of proteins related to oxidative stress and neurodegeneration in an in vitro human blood-brain barrier model［J］.Nanotoxicology,2019,13(2):221-239.
［15］　刘倩,虎亚光,柴尔青.阿司匹林与尼莫地平预处理可改善脑缺血再灌注损伤［J］.解剖学报,2020,51(1):21-25.
［16］　KUMAR M,DADHWAL P,ATREYA H S,et al.Backbone resonance assignments of the dimeric domain of the p50 NF-kappaB subunit［J］.Biomol NMR Assign,2020,14(1):9-11.
［17］　GU X L.MicroRNA-124 prevents H2O2-induced apoptosis and oxidative stress in human lens epithelial cells via inhibition of the NF-κB signaling pathway［J］.Pharmacology,2018,102(3-4):213-222.
［18］　SHI X M,XU G M,ZHANG G J,et al.Low-temperature plasma promotes fibroblast proliferation in wound healing by ROS-activated NF-κB signaling pathway［J］.Curr Med Sci,2018,38(1):107-114.

相似文献/References:

[1]赵红岗 李东亮,王东霞 祈英杰,周素凤,等.黄体酮对大鼠脑缺血区水和钠钾钙含量的影响[J].新乡医学院学报,2000,17(02):077.
[2]张璐璐,朱欣茹,李文强,等.核转录因子 E2 相关因子 2对脑缺血再灌注大鼠脑组织中炎症因子表达和神经胶质细胞活化的影响[J].新乡医学院学报,2020,37(4):301.[doi:10.7683/xxyxyxb.2020.04.001]
　ZHANG Lulu,ZHU Xinru,LI Wenqiang,et al.Expression of nuclear factor erythroid 2-related factor in the brain of ischemia-reperfusion rats and the effect of it′s over expression on the activation of inflammatory factors and neuroglial cells[J].Journal of Xinxiang Medical University,2020,37(3):301.[doi:10.7683/xxyxyxb.2020.04.001]
[3]李　青,白宏英,徐志秀,等.乌司他丁对脑缺血再灌注大鼠脑组织中基质金属蛋白酶-9活性和细胞外基质表达的影响[J].新乡医学院学报,2020,37(10):912.[doi:10.7683/xxyxyxb.2020.10.003]
　LI Qing,BAI Hongying,XU Zhixiu,et al.Effect of ulinastatin on the activity of matrix metalloproteinase-9 and the expression of extracellular matrix in brain tissues of rats with cerebral ischemia-reperfusion[J].Journal of Xinxiang Medical University,2020,37(3):912.[doi:10.7683/xxyxyxb.2020.10.003]
[4]赵　欣,裴科阳,谭　军.腺苷预处理对大鼠局灶性脑缺血再灌注后线粒体活性氧含量及线粒体呼吸链复合体Ⅰ、Ⅲ活性和脑组织总抗氧化能力的影响[J].新乡医学院学报,2019,36(4):305.[doi:10.7683/xxyxyxb.2019.04.002]
　ZHAO Xin,PEI Ke-yang,TAN Jun.Effect of adenosine preconditioning on reactive oxygen species content，activity of respiratory chain complex Ⅰ and Ⅲ of mitochondria and the total antioxidant capacity of brain tissueafter focal cerebral ischemia-reperfusion in rats[J].Journal of Xinxiang Medical University,2019,36(3):305.[doi:10.7683/xxyxyxb.2019.04.002]
[5]王文杰,左　霞,郭俐宏,等.大株红景天注射液对大鼠脑缺血再灌注后血管内皮损伤的影响[J].新乡医学院学报,2016,33(1):026.[doi:10.7683/xxyxyxb.2016.01.008]
　WANG Wen-Jie,ZUO Xia,GUO Li-hong,et al.Effect of rhodiola kirilowii Regel injection on rats with vascular endothelial induced by cerebral ischemia reperfusion[J].Journal of Xinxiang Medical University,2016,33(3):026.[doi:10.7683/xxyxyxb.2016.01.008]

更新日期/Last Update: 2021-03-05