摘要
缺血性脑水肿是脑血管病伴发的最常见的死亡原因,是脑梗死必然且重要的病理过程,对其机制的研究及防治对策,一直是神经科的研究热点之一。其发病机制的主要环节为血脑屏障(BBB)受损。目前对脑水肿的治疗主要有高渗性脱水剂,利尿剂,糖皮质激素以及外科手术减压等,这些治疗虽然能一定程度减轻脑水肿,但都没有解决其发病机制的主要环节即BBB受损的问题。中药及复方对缺血性脑损伤的治疗具有一定的优势,其药理作用的多靶点、多途径性引起了医学界的广泛重视。而醒脑开窍法作为治疗中风的一种重要方法,虽然近年来广泛应用于临床,但一直未能引起足够的重视。
本课题通过文献研究,总结并阐述了缺血性中风急性期使用芳香开窍法的必要性,中医药尤其是麝香和冰片对BBB的作用,脑缺血再灌注后BBB损伤的分子机制。根据导师刘亚敏教授等的工作基础,按线栓法造成大鼠大脑中动脉闭塞,建立局灶性脑缺血再灌注模型,造成缺血性脑水肿,采用电镜、免疫组化、RT-PCR等方法,围绕缺血性脑水肿中血脑屏障(BBB)损伤的主要环节,探讨麝香配伍冰片对脑微血管内皮细胞、基质及胶质细胞足突三个环节的作用,系统地分析麝香配伍冰片对缺血后BBB损伤的影响,阐明其作用的分子机制。为临床合理应用开窍醒脑法治疗急性缺血性中风提供科学的依据,对临床应用也具有重要的指导意义和应用前景。
1文献研究
1.1缺血性中风的病机为脑窍痹阻、神机失用,醒脑开窍是缺血性中风的基本治则。醒脑开窍法是中风急性期的重要方法,应及早使用。使用开窍药不仅可醒脑开窍、恢复神机,且可兼顾化痰、活血、息风止痉、清热等,一举多得。开窍法在治疗偏瘫、失语、痴呆等后遗症方面也有一定的优势。
1.2中医药对BBB的影响目前研究较广泛,芳香开窍药麝香、冰片广泛应用于神经系统疾病,二者透过BBB迅速,麝香可抑制脑缺血后炎症、抗自由基损伤、减轻脑水肿、增加血流量等,对缺血性神经元有保护作用。冰片可改善BBB的通透性,使其他药物能更快更多地进入中枢神经系统发挥作用。
1.3脑缺血再灌注后BBB的超微结构损伤,包括内皮细胞肿胀、吞饮小泡增多、紧密连接开放、基底膜破坏及胶质细胞水肿等。BBB损伤分子机制涉及炎性细胞因子、粘附分子、基质金属蛋白酶、水通道蛋白、自由基、花生四烯酸代谢产物、血管内皮生长因子、纤溶酶、凝血酶等,这些因素相互作用,互为因果。
2实验研究
2.1麝香配伍冰片对缺血再灌注后大鼠病理及BBB超微结构影响的研究
目的:通过观察缺血再灌注后大鼠神经元病理及BBB超微结构变化的影响探讨麝香配伍冰片对BBB的保护作用。方法:建立大鼠局灶性脑缺血再灌注模型,光镜下观察各组动物神经元病理变化,电镜下观察BBB超微结构。结果:麝香配伍冰片可以有效减轻缺血再灌注后神经元坏死、水肿,BBB内皮细胞肿胀、紧密连接疏松、基底膜结构模糊及胶质细胞水肿等。结论:麝香配伍冰片通过上述作用减轻BBB损害、脑水肿,减轻缺血再灌注损伤。
2.2麝香配伍冰片对缺血再灌注后大鼠ICAM-1、LFA-1表达的影响
目的:研究缺血再灌注后BBB内皮细胞ICAM-1、LFA-1表达情况,探讨麝香配伍冰片对BBB内皮细胞的保护作用。方法:RT-PCR方法研究ICAM-1mRNA、LFA-1 mRNA表达,免疫组化方法研究ICAM-1蛋白表达变化。结果:麝香配伍冰片、安宫牛黄丸可显著降低缺血再灌注后脑组织中ICAM-1 mRNA及其配基LFA-1 mRNA的表达(P＜0.01),冰片组各时间段ICAM-1 mRNA和缺血2h再灌注48h时间段的LFA-1mRNA也有明显降低(P＜0.05)。蛋白表达方面,麝香配伍冰片组、安宫牛黄丸组、冰片组ICAM-1蛋白表达量较模型组明显下降(P＜0.05),其中麝香配伍冰片组下降最多。结论:麝香配伍冰片可以降低缺血再灌注后粘附分子ICAM-1及其配基LFA-1的表达,从而减轻炎性反应对BBB的损伤而起脑保护作用。
2.3麝香配伍冰片对缺血再灌注后大鼠MMP-9、TIMP-1表达的影响
目的:研究缺血再灌注后BBB基底膜MMP-9、TIMP-1表达情况,探讨麝香配伍冰片对BBB基底膜的保护作用。方法:RT-PCR方法研究MMP-9 mRNA、TIMP-1 mRNA表达,免疫组化方法研究MMP-9蛋白表达变化。结果:麝香配伍冰片组、安宫牛黄丸组、冰片组各缺血再灌注时间段脑组织MMP-9 mRNA表达均受到明显抑制;与假手术组比较,三组TIMP-1 mRNA表达明显升高,在缺血2h再灌注24h时间段,麝香组也有相同变化趋势。MMP-9蛋白表达方面,麝香配伍冰片组、冰片组、安宫牛黄丸组较模型组表达下降明显(P＜0.05)。结论:麝香配伍冰片可通过抑制MMP-9表达、提升TIMP-1表达而减轻基底膜细胞外基质的降解和BBB通透性。
2.4麝香配伍冰片对缺血再灌注后大鼠AQP4表达的影响
目的:研究麝香配伍冰片对缺血再灌注后BBB胶质细胞AQP-4表达情况,探讨麝香配伍冰片对BBB胶质细胞的保护作用。方法:分别用RT-PCR方法、免疫组化方法研究AQP-4 mRNA、AQP-4蛋白表达的变化。结果:在缺血再灌注各时间段,麝香配伍冰片、安宫牛黄丸、冰片均可明显抑制缺血脑组织中AQP-4 mRNA的表达,其中麝香配伍冰片、安宫牛黄丸组降低幅度最大。蛋白表达方面,麝香配伍冰片组、冰片组、安宫牛黄丸组下降明显(P＜0.05)。结论:麝香配伍冰片通过抑制缺血再灌注后脑组织AQP-4 mRNA和蛋白的表达而降低BBB通透性、减轻脑水肿而起到脑保护作用。
3本研究的创新性
从古至今,绝大多数医家均认为中风急性期出现意识不清才使用“开窍法”,我们以大鼠局灶性脑缺血再灌注模型为基础,研究“芳香开窍”代表药物麝香、冰片药对缺血再灌注后BBB损伤的影响机制,深入研究其作用机理,为临床治疗缺血性中风即使未出现意识不清也可应用芳香开窍法提供依据。
过去对麝香、冰片多进行单味药的药理研究,我们以成分比较清楚的、且能通过BBB的两个单味中药麝香、冰片配伍组合进行系统的研究,有别于传统的大复方研究,不仅有利于药物作用的分析及其应用时增效减毒作用的研究,而且便于新药的开发,具有潜在的应用价值。
探讨麝香配伍冰片抗缺血性脑损伤的作用机理,对揭示该药对配伍规律及其“芳香开窍”作用机理具有理论意义。现有的研究未能系统阐明该药对抗缺血性脑损伤的机制,因而未能较清晰地认识其作用环节,限制了该药对临床的合理使用。本研究从多角度探讨该药对配伍抗脑缺血损伤的作用及机理。这对于深入认识该药对的配伍原理和临床合理应用均具有指导作用。
围绕缺血性脑水肿中BBB损伤的主要环节,从内皮细胞损伤、基质破坏和胶质细胞损伤三个环节动态研究麝香配伍冰片抗缺血性脑水肿的作用和分子机制,从而系统地研究其作用环节。
Ischemic brain edema is the most common cause of death following cerebrovascular disease.It is the necessary and important pathological process.It is one of the focal points in neurological territory.Injuried Blood Brain Barrier(BBB) is the major pathogenesy.Treatments on brain edema currently are hyperisotonic dehydrater,diuretic,glucocorticoid and surgical intervention to reduce intracranial pressure.Although these treatments could relieve brain edema to a certain degree,its major pathogenetic link that BBB is injured is still remain to be resolved.Traditional Chinese drugs and complex prescriptions have certain advantages on treating ischemic brain injuries,it has aroused extensive attention because of the multi-target, multi-channel pharmacological effects.As an important tool of treating stroke, brain waking-up therapy has not been able to arouse enough attention although in recent years is widely Used in clinical.
Through studying literatures,this project summed up and expounded the necessity of using resuscitation-inducing therapy with fragrant drugs during the acute stage of ischmic stroke,traditional Chinese Medicine on the role of BBB,particularly the musk and borneol,the molecular mechanism of BBB injury after cerebral ischemia-reperfusion.According to the basis of the work of Professor Liu Yamin,we established the focal cerebral ischemia-reperfusion model by suture middle cerebral artery occlusion in rats.Then Focusing on ischemic brain edema in the BBB injury of the major aspects,we Explored the influence of Borneol with musk on cerebral microvascular endothelial cells, stromal cells and glial foot using electron microscopy,immunohistochemistry, RT-PCR and other methods.The role of musk with Borneol BBB and its molecular mechanism after ischemic injury was systematicly analyzed.This could provide scientific basis for clinical treating acute ischemic stroke with brain waking-up therapy.
1 Documents study
1.1 Obstruction in the brain orifice and its functions failing to work is the leading pathogenesis for ischemic stroke,so brain waking-up therapy is the basic rule to cure ischemic stroke.It is an an important means of acute phase of stroke and should be used as soon as possible.Resuscitation-inducing drugs could wake up brain to resume its functions,eliminate sputum,promote blood flow,relieve spasm by calming endogenous wind,clear heat and so on. Resuscitation-inducing therapy also has advantages when treating hemiparalysis,anepia,dementia and other residuals.
1.2 The impact of Traditional Chinese Medicine on BBB is researched widely right now.Musk and borneol are widely used in diseases of the nervous system,they could traverse BBB quickly.Musk could restrain inflammation, resist injuries because of free radical,extenuate brain edema and increase blood flow after brain ischemia.Borneol could improve permeability of BBB so as to help other drugs enter central nervous system more and fasterly.
1.3 The ultrastructure injury of BBB after cerebral ischemia-reperfusion includes endothelial cell swelling,increased pinocytotic vesicles,tight junctions open,membrane damage and edema and glial cell swelling and so on. The molecular mechanisms of BBB injury involves inflammatory cytokines, adhesion molecules,matrix metalloproteinases,water channel proteins,free radicals,arachidonic acid metabolites,vascular endothelial growth factor, plasminogen,thrombin,etc.
2 Experimental research
2.1 Studies about the influence of Musk with Borneol on pathology and BBB ultrastructure after ischemia-reperfusion
Objective:Discuss the protection effect of Musk with borneol on BBB by observation of neuron pathology and BBB ultrastructure after ischemia-reperfusion.Methods:Focal cerebral ischemia-reperfusion model(MCAO) rat was set up first of all,then observation of neuron pathology using light microscopy and BBB ultrastructure using electron microscopy was done.Results:Musk with borneol could effectively reduce neuronal necrosis, edema,BBB endothelial cell swelling,loose tight junctions,basement membrane structure and glial cell line-fuzzy edema after ischemia-reperfusion.
Conclusion:Musk with Borneol reduced BBB damage,brain edema and ischemia-reperfusion injury through these mentioned roles above.
2.2 Studies about the influence of Musk with Borneol on the expression of ICAM-1 and LFA-1 after ischemia-reperfusion
Objective:Discuss the protection effect of Musk with borneol on BBB endothelial cell by investigating the expression of ICAM-1 and LFA-1 on the rats with focal cerebral ischemia-reperfusion injury.Methods:ICAM-1 and LFA-1 mRNA expression were measured utilizing real-time PCR,ICAM-1 protein expression was evaluated utilizing immunohistochemistry.Results:ICAM-1 mRNA,LFA-1 mRNA expression in musk with borneol group and Angong Niuhuang Pill group significantly decreased(P＜0.01),ICAM-1 mRNA expression each time and LFA-1 mRNA expression after reperfusion for 48h in borneol group also significantly reduced(P＜0.05).ICAM-1 protein expression in musk with borneol group,Angong Niuhuang Pill group and borneol group decreased significantly compared with model group(P＜0.05),it decreased at most in musk with borneol group.Conclusion:Musk with borneol could protect BBB against inflammatory response through reducing the expression of adhesion molecule ICAM-1 and its ligand LFA-1 after ischemia-reperfusion.
2.3 Studies about the influence of Musk with Borneol on the expression of MMP-9 and TIMP-1 after ischemia-reperfusion
Objective:Discuss the protection effect of Musk with borneol on BBB basement membrane by investigating the expression of MMP-9 and TIMP-1 on the rats with focal cerebral iscbemia-reperfusion injury.Methods:MMP-9 and TIMP-1 mRNA expression were measured utilizing real-time PCR,MMP-9 protein expression was evaluated utilizing immunohistochemistry.Results:MMP-9 mRNA expression in musk with borneol group,borneol group and Angong Niuhuang Pill group were significantly inhibited(P＜0.01).TIMP-1 mRNA expression significantly increased compared with the sham-operated group in those three groups and musk group after 24h reperfusion.MMP-9 protein expression in musk with borneol group,Angong Niuhuang Pill group and borneol group decreased significantly compared with model group(P＜0.05).Conclusion:Musk with borneol could reduce the degradation of extracellular matrix basement membrane and BBB permeability by inhibiting the expression of MMP-9 and elevating the expression of TIMP-1.
2.4 Studies about the influence of Musk with Borneol on the expression of AQP-4 after ischemia-reperfusion
Objective:Discuss the protection effect of Musk with borneol on BBB glial cell by investigating the expression Of AQP-4 on the rats with focal cerebral ischemia-reperfusion injury.Methods:The expression of AQP-4 mRNA were measured utilizing real-time PCR and its protein expression was evaluated utilizing immunohistochemistry.Results:AQP-4 mRNA expression in musk with borneol group,borneol group and Angong Niuhuang Pill group were significantly inhibited,it decreased at most in musk with borneol group and Angongniuhuang Pill group decreased the largest.AQP-4 protein expression in musk with borneol group,Angong Niuhuang Pill group and borneol group decreased significantly compared with model group(P＜0.05).Conclusion:Musk with Borneol could reduce the BBB permeability and brain edema through inhibiting the expression of AQP-4 mRNA and protein after ischemia-reperfusion so as to protect the brain.
3 The contributions of this research are as follows
From ancient times,the vast majority of doctors are of the view that the resuscitation-inducing therapy should be used when conscious disturbance appears at the acute phase of stroke.Musk with borneol is the type of fragnant drugs for inducing resuscitation.We set up the focal cerebral ischemia-reperfusion rat model to study the influence of musk with borneol on BBB injury after ischemia-reperfusion and research deeply its action mechanism.We hope to provide a basis for using resuscitation-inducing therapy with fragrant drugs even if conscious disturbance has not appeared when treating ischemic stroke.
In the past,most researches on musk and borneol were one-flavor drug pharmacological study.Having clear constituents and the ability to traverse BBB,musk with borneol was studied Systematically,which is different from the traditional research on large compound.That has potential application value not only because of the advantages to the analysis of drug effect and researches on synergy and attenuation when applied,but also the facility for the development of new drugs.
Discussion on the mechanism of musk with borneol against ischemic brain damage has theoretical significance to reveal its compatibility law and mechanism of resuscitation-inducing therapy with fragrant drugs.The mechanism against ischemic brain damage of musk with borneol has not been illuminated in existing researches and the link of its role has not been clearly known,so its clinical application has been restrained.This study discussed the mechanism of musk with borneol against ischemic brain damage from different perspectives:It has guiding role to recognize its compatibility law and apply reasonablly.
Around Ischemic brain edema around the BBB injury in the main aspect of BBB injury during ischemic brain edema,the effect against ischemic brain edema and molecular mechanism of musk with borneol was researched dynamically from endothelial cells damage,matrix damage and glial cell injury to study the link of its role systematically.
引文
[1]Kay R,Woo J,Kreel L,et al:Sroke subtypes among chinese living in Hong Kong:the Shatin Stroke Registry.Neurology,1992;4(2):985-987
[2]Ter Minassian A.Treatment of cerebral oedema.Ann Fr Anesth Reanim,2003;Apr,22(4):336-48
[3]刘新峰.急性卒中脱水治疗的争论与实施.国外医学:脑血管疾病论坛,2003;1(1):6-8
[4]Pardridge WM.Non-invasive drug delivery to the human brain using endogenous blood-brain barrier transport systems.Pharmaceutical Science and Technology Today,1999;2:49-59
[5]李瑾翡,王宁生,陆惠文,等.麝香酮药代动力学研究.中药新药与临床药理,2000;11(4):208-210
[6]蒋振亚,李常度,周东,等.麝香对大鼠实验性脑缺血神经元损伤的保护作用.中国中医药科技,2001;8(2):97-98
[7]赵保胜,宓穗卿,张银卿.冰片对大鼠脑微血管ICAM-1表达的影响.中药新药与临床药理,2001;12(2):88
[8]刘亚敏,张赐安,徐秋英,等.全脑缺血再灌注大鼠脑组织水通道蛋白-4 mRNA 的表达及药物对其影响.中国中医急症,2002;12(6):475-477
[9]张赐安,刘亚敏,徐秋英,等.芳香开窍法对全脑缺血再灌注大鼠脑组织NO 含量及eNOS、iNOS表达的影响.广州中医药大学学报,2002;(2):31-35
[10]沈强,刘亚敏,张赐安,等.麝香、冰片对全脑缺血再灌注大鼠脑微血管内皮细胞ICAM-1表达的影响.中西医结合心脑血管病杂志,2003;1(3):136
[11]刘亚敏,张赐安,林培政,等.芳香开窍法治疗急性缺血性中风的临床观察.广州中医药大学学报,2002:19(3):165-168
[12]刘亚敏,夏鑫华,赵光峰,等.麝香配伍冰片对局灶性脑缺血再灌注大鼠脑含水量及血脑屏障通透性的影响.广州中医药大学学报,2007;24(6):498-501
[13]刘亚敏,夏鑫华,赵光峰,等.麝香配伍冰片对局灶性脑缺血再灌注大鼠行为学及脑梗死体积的影响.中西医结合心脑血管病杂志,2006;4(7):590-591
[14]方永奇,魏刚,吴启端,等.石菖蒲醒脑丌窍作用物质基础及其作用机制研究.医学研究杂志,2007;36(9):60
[15]王永炎.中医内科学.第1版.上海:上海科学技术出版社,1997:117-156,344-349
[16]颜冰,黄培堂.血脑屏障与脑药物转运.生物技术通讯,2002;13(3):199-201
[17]唐洪梅,李锐.石菖蒲通过大鼠血脑屏障的化学成分分析.中医药研究,2002;18(1):401
[18]方永奇,魏刚,柯雪红.GC-MS分析石菖蒲挥发油透大鼠血脑屏障的成分研究.中药新药与临床药理,2002;13(3):181-182
[19]陈春富,郭述苏,冯林,等.黄芪对大鼠脑缺血血脑屏障及脑血流的影响.中风与神经疾病杂志,1998;5(3):132-134
[20]刘建辉,冀风云,王婷,等.三七总皂甙对实验性脑缺血脑血流及血脑屏障的影响作用.河北医药,2002;24(14):249-250
[21]罗玉敏,秦震.黄芪甲苷对大鼠局灶脑缺血血脑屏障破坏的影响.中国临床神经学,2000;8(4):280-281
[22]张素平,何锐,王慕真,等.活血化瘀药抗大鼠脑缺血、再灌注想血脑屏障损伤的保护作用.中国医院药学杂志,2003;23(10):579-581
[23]乔晋,杨养贤,陈明霞,等.活血效灵丹对脑出血兔血脑屏障及脑组织超微结构的影响.西安医科大学学报,2002;23(3):302-305
[24]李克玲,黄启福,邹丽琰,等.醒脑健神胶囊对家兔脑血肿及血脑屏障影响的实验研究.中风与神经疾病杂志,1997;14(1):21-23
[25]童建国,王先荣,朱刚,等.活血化瘀中药对脑挫伤的治疗作用及其机制的实验研究.第三军医大学学报,2001;23(7):8211
[26]谭峰,贺石林,虞佩兰,等.补阳还五汤对大白鼠血脑屏障影响的实验研究.中国中医急症,1993;2(5):2231
[27]刘晓霓,李月玲,孙成,等.针刺对实验性脑缺血家兔血脑屏障超微结构及血液流变学的影响.牡丹江医学院学报,2002;23(2):1-4
[28]纪晓军,何宏,温兆霞,等.头穴针刺对大鼠急性脑出血脑屏障影响的实验研究.中国急救医学,2001;21(11):624-625
[29]D,DU,Wu Gc,et al:Effect of electroacupuncture on blood-brain barrier after cerebral ischemia reperfusion in rat.Acupuncture Electro Ther2apeutics Res 2001,26(122):11
[30]莫启忠,宫斌,茅惠明,等.H-麝香酮在体内的吸收分布和排泄:中药麝香芳香开窍机制的初步探讨.中成药研究,1984;(7):1
[31]梁辉,陈虎,高颖,等.麝香酮抗局灶性脑缺血损伤的实验研究.中成药,2003;25(3):226-227
[32]刘亚敏,张赐安,徐秋英,等.芳香开窍法对全脑缺血再灌注大鼠脑水肿及超氧化物歧化酶、丙二醛水平的影响.中国中医药信息杂志,2002;9(7):22-24
[33]刘亚敏,张赐安,徐秋英,等.麝香、冰片对全脑缺血再灌注大鼠脑组织氨基酸类神经递质的影响.中药新药与临床药理,2002;13(4):231-233
[34]沈强,刘亚敏,张赐安等.麝香与冰片对全脑缺血再灌注大鼠脑组织白细胞介素-1β mRNA表达的影响.时珍国医国药,2003:14(7):385-386
[35]李野,解建国.化瘀利水、醒脑开窍法对脑梗塞兔脑细胞线粒体Ca~(2+)、Mg~(2+)的影响.首都医药,2004;(14):31-32
[36]吴寿荣,程刚,冯岩.冰片药理作用的研究进展.中草药,2002;32(12):1143-1144
[37]王宁生,梁美容,刘启德,等.冰片“佐使则有功”之实验研究.中医杂志,1994;35(1):461
[38]梁美容,叶少梅,张银卿,等.冰片对兔、大鼠脑组织伊文氏蓝染色作用的观察.广州中医药大学学报,1993;10(4):2111
[39]徐伟,宗锐.薄荷醇及冰片对磺胺嘧啶和伊文思蓝在脑中分布的影响.中药药理与临床,1995;11(6):311
[40]董先智,汤小爱,高秋华,等.冰片对顺铂透血脑屏障促进作用的研究.中国药学杂志,2002;37(4):751
[41]黄萍,吴清和,荣向路,等.冰片与川芎配伍抗脑缺血再灌注损伤作用机理的研究.广州中医药大学学报,2001;18(4):332
[42]林咸明,陈华德,严建伟,等.穴位注射“冰片液”对血脑屏障通透性的影响.针刺研究,2003;28(2):99-101
[43]李东晓,邓文龙.冰片促透作用研究进展.中药药理与临床,2007;23(1):87-89
[44]刘煜德,孙丹邈,黄小洵,等.冰片对BMSCs透过血脑屏障的影响.亚太传统医药,2007;3(7):46-48
[45]刘洋,张伯礼,胡利民.冰片的药理实验研究概况.天津中医药,2003;20(4):85-87
[46]赵保胜,徐勤,宓穗卿,等.冰片促血脑屏障开放与一氧化氮含量改变的关系初探.脑与神经疾病杂志,2001;9(4):207-209
[47]赵保胜,徐勤,宓穗卿,等.冰片促血脑屏障开放与病理性开放的比较.中药新药与临床药理,2002;13(5):287-288
[48]刘煜德,黄小洵,孙丹邈.开窍药对血脑屏障作用研究进展.亚太传统医药,2007;3(6):32-34
[49]袁卓,张军平,刘养凤,等.P-糖蛋白与冰片促血脑屏障生理性开放的关系探讨.天津中医药,2006;23(3):261-262
[50]陈艳明,王宁生.冰片对P-糖蛋白的影响.中药新药与临床药理,2003;14(2):96-99
[51]蒲传强,匡培根,金泽武道,等.脑缺血后血脑屏障超微结构的研究.中华神经精神科杂志,1994;27(2):67-68
[52]Lacaz-Viera F,Jaeger MM.Protein kinase inhibitors and the dynamics of tight junction opening and closing in A6 cell monolayers.J Membrane Biol 2001;184:185-96
[53]石向群,杨金升,张志琳,等.水通道蛋白4基因干预对大鼠脑缺血再灌注损伤后血脑屏障形态及功能的影响.中国临床康复,2004;8(31):6928-6930
[54]吴松笛,万琪,夏峰,等.大鼠急性脑缺血再灌注后血脑屏障通透性的超微结构与血清S100B蛋白水平的动态变化.第四军医大学学报,2006;27(10):902-904
[55]杨允东,关宁,罗俊生.大鼠脑缺血模型血脑屏障结构和功能状态的研究.锦州医学院学报,2005;26(5):23-26
[56]李玲,黄如训,苏镇培.局部脑梗塞再灌流边缘区血脑屏障超微结构改变.中风与神经疾病杂志,1998:15(1):11-13
[57]李曼霞,董志.脑缺血再灌注时血脑屏障损伤的炎性机制研究进展.解放军药学学报,2007;23(1):60-62
[58]海舰,丁美修.脑缺血再灌注时血脑屏障损伤的炎性机制.国外医学脑血管疾病分册,2001;9(1):15-17
[59]Rostenberg GA,Estrada BS,Dencoff JE.Matrix metalloproteinases and TIMPs are associated with Blood-brain barrier opening after reperfusion in rat brain.Stroke,1998,29(10):2189-2195
[60]Rivera S,Ogier C,Jourquin Jet al.Gelatinase B and TIMP-1 are regulated in a cell-and time-dependent manner in association with neuronal death and glial reactivity after global forebrain ischemia.Eur J Neurosci.2002 Jan;15(1):19-32
[61]Xiao F.Bench to bedside:brain edema and cerebral resuscitation:the present and future.Acad Emerg Med,2002,9(9):933-946
[62]Nicchia GP,Rossi A,Mola MG,et al.Actin cytoskeleton remodeling governs aquaporin-4 localization in astrocytes.Glia,2008,56:1755-1766
[63]KE c,Poon WS,Ng HK,et al.Heterogeneous responses of aquaqorin-4 in edema formation in a replicated severe traumatic brain injury model in rats.Neurosci Lett,2001,301(1):21-24
[64]Kiening KL,van Landeghem FK,Schreiber S,et al.Decreased hemispheric aquaporin-4 is linked to evolving brain edema following controlled cortical impact injury in rats.NeurosciLett,2002,324:105-108
[65]Li C,Hirooka Y,Honda R,et al.Distribution ofaquaporin-9 in the rat:an immunohistochemical study.Int J Tissue React,2005,27(2):51-58
[66]李燕华,罗永坚,李吕力,等.脑缺血后AQP9表达与血脑屏障通透性的关系.中风与神经疾病杂志,2008;25(2):33-35
[67]Prerrerkom T,Wiessner C,Allegrini PR,et al.Plasminogen activator in experimental permanent focal cerbral ischemia.Brain Res,2000,882(1-2):19-25
[68]王剑峰,刘艳丽.脑缺血再灌注后影响血脑屏障通透性的因素.国际脑血管病杂志,2006;14(12):930-932
[69]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats.Stroke,1989;20:84-91.
[70]Tibo Gerriets MD,Erwin Stolz MD,Maureen Walberer,et al.Complications and pitfalls in rat stroke models for middle cerebral artery occlusion:A comparison between the suture and the macrosphere model using magnetic resonance angiography.Stroke,2004;35:2372-2377
[71]Dietrich JB.The adhesion molecule ICAM-1 and its regulation in relation with the blood-brain barrier.J Neuroimmunol.2002 Jul;128(1-2):58-68
[72]Yenari MA,Kunis D,Sun GH,Onley D Hu23F2G,an antibody recognizing the leukocyte CD11/CD18 integrin,reduces injury in a rabbit model of transient focal cerebral ischemia.Exp Neurol.1998 Oct;153(2):223-33
[73]Rothlein R.Overview of leukocyte adhesion.Neurology,1997;49(4):3-4
[74]Zoppo GT,Schinid-Schonbein GW,Mori E,et al.Polymor phonuclear leukocyte occlude capillaries following middle cerebral artery occlusion and reperfusion in baboons.Stroke,1991;22:1276-1283
[75]卫生部疾病预防控制局,中华医学会神经病学分会.中国脑血管病防治指南.第1版.北京:人民卫生出版社,2007:1-2
[76]Harkness KA,Adamson P,Sussman JD,et al.Dexamethasone regulation of matrix metallopro teinase expression in CNS vascular endothelium.Brain,2000,1123:698-709
[77]Yoneda K,Yamamoto N,Asai K,et al.Regulation ofaquaporin-4 expression in astrocytes.Brain Res Mol Brain Res.2001 Apr 18;89(1-2):94-102
[78]李晓宾,董瑞国.AQP4与脑水肿研究进展.神经疾病与精神卫生,2005;5(1):77-79
[79]Manley GT,Fujimura M,Ma T,et al.Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke.Nat Med,2000,6(2):159-163
[80]王志芳.安宫牛黄丸在颅脑损伤及脑卒中病人中应用76例总结.中华临床医学研究杂志,2007;13(22):3193-3194
[81]李净,王键,益气活血法改善气虚血瘀证局灶性脑缺血再灌注模型鼠生物学特征的有效性评价,中国中医基础医学杂志,2003;9(4):22-26
[82]雷奕祺,袁明远,史东葵.醒脑静注射液与安宫牛黄丸治疗脑梗塞急性期临床对照研究.中国中医急症,2002;11(5):359-360
[1]崔高宇.BBB结构研究近况.国外医学脑血管疾病分册,1997;5(5):277-280
[2]邵小鹏,刘勇.川芎嗪对大鼠局部脑缺血星形胶质细胞的影响.交通医学,2001;15(4):362-363
[3]杨堃,蒋先惠,王荣亮等.银杏叶提取物对冷冻所致兔脑水肿的防治作用.福建中医药,2001;32(6):42-43
[4]刘建辉,冀凤云,王婷等.三七总皂甙对实验性脑缺血脑血流及血脑屏障的影响作用.Apoplexy and Nervous Diseases,2002,19(3):164
[5]闫彦芳,张壮,孙塑伦等.三七总皂苷及其主要成分对血管内皮细胞缺氧损伤的保护作用.中国实验方剂学杂志,2002;8(1):34-37
[6]王文安,周永炜,程洁等.三七皂甙对局灶性脑缺血再灌注后MMP-9表达的影响.上海第二医科大学学报,2004;24(9):731-733
[7]何蔚,陈汇,曾繁典.灯盏花素对脑缺血再灌注损伤的保护作用和治疗时间窗.中国临床药理学与治疗学,2003;8(3):278-280
[8]王贤军,夏青,蔡洪信等.白花丹参叶制剂对局灶性脑梗死HSP70及细胞凋亡的影响.中西医结合心脑血管病杂志,2005;3(2):140-141
[9]李克玲,黄启福,朱陵群等.中风脑得平冲剂对实验性兔脑血肿的作用.北京中医药大学学报,1996;19(4):22-24
[10]童建国,王宪荣,朱刚等.活血化瘀中药对脑挫裂伤的治疗作用及其机制的实验研究.第三军医大学学报,2001;23(7):821-823
[11]乔晋,杨养贤,陈明霞等.活血效灵丹对脑出血兔BBB及脑组织超微结构的影响.西安医科大学学报,2002,23(3):302-305
[12]张素平,何锐,王慕真等.活血化瘀药抗大鼠脑缺血-再灌注性血脑屏障损伤的保护作用.中国医院药学杂志,2003;23(10):579-581
[13]邓婉青,何锐,张素平.活血化瘀药对大鼠脑缺血再灌注血管源性脑水肿的影响.中国临床康复,2004;8(10):1914-1915
[14]王慕真,张素平,何锐.活血化瘀汤预处理对大鼠局灶缺血再灌注损伤的保护作用.广东药学院学报,2004;20(3):255-257
[15]崔向宁,尹岭,王玉来.脑疏宁对颅脑创伤后大鼠血脑屏障通透性及MMP-9表达的影响.北京中医药大学学报,2005;28(2):60-62
[16]谭峰,欧正武,李安国等.补阳还五汤对脑水肿的钙拮抗作用.中国危重病急救医学,1995;7(3):136-137
[17]陈春富,郭述苏,冯林等.黄芪对大鼠脑缺血血脑屏障及脑血流的影响.卒中与 神经疾病杂志,1998;5(3):132-134
[18]杨志敏,周文斌,黄培新等.益气活血药对脑缺血再灌注模型脑水肿及超微结构的影响.广州中医药大学学报,1999;16(2):114-118
[19]邱根全,石秦东,蔡云等.益气活血开窍复方对缺血再灌注脑组织兴奋性氨基酸及超微结构的影响.西安交通大学学报(医学版),2002;23(5):489-492
[20]王彦田,王鑫国,张会珍.益气活血复方对缺血再灌注损伤性痴呆大鼠脑微血管内皮细胞一氧化氮合酶表达的影响.中国中医基础医学杂志,2004;10(8):27-29
[21]沈淑萍,朱莉萍,孙佳红等.益气活血中药对大鼠前脑不全缺血再灌注后海马区血管内皮细胞生长因子表达的影响.包头医学院学报,2004;20(4):278-280
[22]张小婷,张少君.益气活血药对脑缺血再灌注脑组织兴奋性氨基酸的实验研究.医学文选,2004;23(5):572-573
[23]俞浩,王钦茂,周世文.参芪提取物对实验性小鼠闭合性脑外伤的影响.中国兽药杂志,2005;39(3):16-18
[24]张树泉,徐西元,王凤霞等.益气祛瘀化痰方对缺血再灌注鼠脑损伤的抗氧化作用研究.中西医结合心脑血管病杂志,2005;3(5):414-415
[25]白海波,万海同,严伟民等.养阴益气活血冲剂对大鼠脑缺血再灌注氧自由基损伤的保护作用.中国中医急症,2001;10(2):96-97
[26]万海同,白海波,杨洁红等.养阴益气活血冲剂对脑缺血再灌注损伤保护作用的实验研究.中国中西医结合急救杂志,2001;8(2):85-87
[27]邓奕辉,陈大舜,李定祥.滋阴益气活血法对糖尿病合并脑缺血大鼠内皮素、一氧化氮及一氧化氮合酶含量的影响.中国中医药信息杂志,2001;8(8):25-26
[28]言惠文,邬力祥,刘发益等.川芎嗪、参麦注射液对缺氧复氧损伤后鼠脑星形胶质细胞释放一氧化氮的影响.湖南中医学院学报,2002;22(2):21-23
[29]李国庆,武继涛,王同聚等.复方仙灵脾注射液对大鼠局灶性脑缺血血流变和自由基的影响.中医药研究,2002;18(2):33-35
[30]武继涛.复方仙灵脾注射液对全脑缺血再灌注大鼠脑组织含水量、血清TNF-α、IL-1、IL-6含量的影响.中医药学刊,2003:21(11):1904,1905,1910
[31]周清安,李建生,王至婉等.补肾活血及益气活血方药对脑缺血老龄大鼠Ca~(2+)和Ca~(2+)-ATPase的影响.中国中医急症,2004;13(11):756-757
[32]周清安,李建生,王至婉等.补肾活血、益气活血方药对脑缺血老龄大鼠脑电图和脑含水量的影响.中国中医急症,2004;13(12):827-828
[33]周清安,李建生,王至婉等.补肾活血与益气活血方药对脑缺血老龄大鼠白介素-6和转化生长因子的影响.中国中医急症,2005;14(6):562-563
[34]黄世敬,黄启福,孙塑伦等.救脑宁注射液对实验性脑出血大鼠脑水肿及脂质过 氧化的影响.北京中医药大学学报,1998;21(2):38-40
[35]王桐生,龙子江,陈文俊.脑创宁对闭合性脑外伤模型小鼠血脑屏障的保护作用.中药药理与临床,2002;18(6):41-42
[36]徐伟,王宗锐.薄荷醇及冰片对磺胺嘧啶和伊文思蓝在脑中分布的影响.中药药理与临床,1995;(6):31-33
[37]陈艳明,王宁生.冰片对血脑屏障体外模型细胞间紧密连接和细胞吞饮囊泡的影响.中国中西医结合杂志,2004;24(7):632-634
[38]赵保胜,徐勤,宓穗卿等.冰片促血脑屏障开放与一氧化氮含量改变的关系初探.脑与神经疾病杂志2001;9(4):207-209
[39]赵保胜,宓穗卿,张银卿.冰片对大鼠脑微血管内皮细胞ICAM-1表达量的影响.中药新药与临床药理,2001;12(2):88-90
[40]赵保胜,刘启德.冰片促血脑屏障开放与病理性开放的比较.中药新药与临床药理,2002;13(5):287-288
[41]李野,解建国.化瘀利水,醒脑开窍法对脑梗塞兔脑细胞线粒体Ca2+,Mg2+的影响.首都医药,2004;(14):31-32
[42]张赐安,刘亚敏,徐秋英等.芳香开窍法对全脑缺血再灌注大鼠脑组织NO含量及NO合酶表达的影响.广州中医药大学学报,2002;19(2):115-118
[43]刘亚敏,张赐安,徐秋英等.芳香开窍法对全脑缺血再灌注大鼠脑水肿及超氧化物歧化酶、丙二醛水平的影响.中国中医药信息杂志,2002;9(7):22-24
[44]刘亚敏,张赐安,徐秋英等.麝香、冰片对全脑缺血再灌注大鼠脑组织氨基酸类神经递质的影响.中药新药与临床药理,2002;13(4):231-233
[45]刘亚敏,张赐安,徐秋英等.全脑缺血再灌注大鼠脑组织水通道蛋白-4 mRNA 的表达及药物对其影响.中国中医急症,2002;11(6):474-475
[46]沈强,刘亚敏,张赐安.麝香、冰片对全脑缺血再灌注大鼠脑微血管内皮细胞ICAM-1表达的影响.中西医结合心脑血管病杂志,2003;1(3):136-137
[47]沈强,刘亚敏,张赐安等.麝香与冰片对全脑缺血再灌注大鼠脑组织白细胞介素-1β mRNA表达的影响.时珍国医国药,2003;14(7):385-386
[48]邓淑琴,梁军红,严红.“利脑灵”对局部冷冻法所致脑水肿作用的研究.天津中医学院学报,1994;(2):35-36
[49]戴春福.清营汤降低家兔脑脊液乳酸脱氢酶的观察.福建中医学院学报,1998;8(4):30-31
[50]郭瑞友,张苏明,方思羽等.海风藤提取物对老龄大鼠缺血性脑损伤保护作用研究.华中医学杂志,2000;24(4):177-178
[51]郭瑞友,于义英,方思羽等.海风藤对局灶性脑缺血治疗作用的实验研究.中西医结合心脑血管病杂志,2003;1(8):461-463
[52]梅建勋,张伯礼,陆融.清脑益智方对谷氨酸损伤神经元保护作用的体外实验观察.中药新药与临床药理,2001;12(4):252-255
[53]刘泰,唐宇平,曾祥发等.健神利水Ⅰ号对血管源性脑水肿影响的实验研究.广西中医药,2002;25(6):50-52
[54]王中琳.养血清脑颗粒对MCAO模型大鼠神经功能保护机制研究.辽宁中医杂志,2002;29(12):704-706
[55]唐涛,罗团连,黎杏群等.中药脑溢安改善脑出血大鼠神经元有氧代谢的药效机制.中国临床康复,2003;7(16):2288-2289
[56]何纲,刘茂才,黄培新等.脑脉Ⅱ号口服液对高血压脑出血大鼠脑损伤的保护作用.中国中西医结合急救杂志,2005;12(1):13-16
[57]王键,郜峦,胡建鹏.3种中医治法对脑缺血再灌注损伤大鼠自由基和神经元凋亡的动态比较研究.中国实验方剂学杂志,2005;(5):49-51