Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats
Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats
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DOI：
10.1016/s0006-8993(96)00815-3
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发表时间：
1996-11-11
期刊：
影响因子：
2.9
通讯作者：
Ginsberg, MD
中科院分区：
文献类型：
--
作者：
Belayev, L;Busto, R;Ginsberg, MD
采用灵敏定量荧光法观察脑中动脉闭塞（MCAo）后血脑屏障（BBB）开放的时间和区域模式。雄性Sprague-Dawley大鼠用氟烷麻醉后，逆行插入涂有聚l -赖氨酸的腔内尼龙缝合线，经颈外动脉进入颈内动脉和MCA，进行2小时的临时MCAo。在MCAo发作后2、3.24或48小时，通过Evans蓝（EB）染料外渗来判断血脑屏障的损伤。1或2 h后对6个脑区进行EB荧光定量。72h时用组织病理学材料定量测定脑梗死体积。在MCAo 3小时后，同侧尾核和新皮层的EB外渗首先明显可见，24-26小时不明显，48-50小时最大。荧光定量证实，在2-3小时没有血脑屏障开口，但在以后的所有时间都存在。在MCAo的同侧半球，与假手术大鼠相比，4小时时EB外渗平均增加179%，5小时时增加407%，26小时时增加311%，50小时时增加264%（每种情况下，与假手术相比P < 0.05）。72h梗死组织体积为163+/-7.7 mm(3)。我们的研究结果表明，初始的急性血脑屏障破坏发生在MCAo后3 - 5小时，随后，更广泛的区域血脑屏障通透性增加出现在48小时。Evans Blue外渗的区域测量提供了定量局灶性脑缺血血脑屏障破坏的精确方法；该方法在评估药物对血脑屏障的保护特性方面具有相当大的实用价值。
A sensitive quantitative fluorescence method was used to explore the time course and regional pattern of blood-brain barrier (BBB) opening after transient middle cerebral artery occlusion (MCAo). Male Sprague-Dawley rats were anesthetized with halothane and subjected to 2 h of temporary MCAo by retrograde insertion of an intraluminal nylon suture, coated with poly-L-lysine, through the external carotid artery into the internal carotid artery and MCA. Damage to the BBB was judged by extravasation of Evans Blue (EB) dye, which was administered either 2, 3.24 or 48 h after onset of MCAo. Fluorometric quantitation of EB was performed 1 or 2 h later in six brain regions. Cerebral infarction volumes were quantitated from histopathological material at 72 h. EB extravasation first became grossly visible in the ipsilateral caudoputamen and neocortex following 3 h of MCAo, was grossly unapparent at 24-26 h, and was maximal at 48-50 h. Fluorescence quantitation confirmed that BBB opening was absent at 2-3 h but present at all later times. In the hemisphere ipsilateral to MCAo, a 179% mean increase in extravasation of EB (compared to sham rats) was measured at 4 h, 407% at 5 h, 311% at 26 h adn 264% at 50 h. (in each case, P < 0.05 vs. sham). The volume of infarcted tissue at 72 h in this model was 163+/-7.7 mm(3). Our results indicate that an initial, acute disruption of the BBB occurs between 3 an 5 h following MCAo, and that a later, more widespread increase in regional BBB permeability is present at 48 h. Regional measurement of Evans Blue extravasation offers a precise means of quantitating BBB disruption in focal cerebral ischemia; this method will be of considerable utility is assessing the BBB-protective properties of pharmacological agents.