这项工作得到了国家自然科学基金（82173781和82373835）、博士后科研项目（BKS212055）、佛山市科学技术局科技创新项目（2320001007331）、广东省基础与应用基础研究基金（2019A1515010806）、广东省通用高校重点领域项目（智能制造）（2020ZDZX2057）和通用科研项目（特色创新）的支持。广东省大学排名 （2019KTSCX195）.

实验方案是按照成都中医药大学实验动物使用和机构动物护理和使用委员会指南（记录编号：2019-DL-002）进行的。所有动物研究数据均按照 ARRIVE（动物研究 ：体内实验 报告）指南进行记录。本研究使用体重 250 g ± 20 g 且年龄为 6-8 周龄的雄性 Sprague Dawley （SD） 大鼠。有关所用动物、试剂和设备的详细信息列在 材料表中。
1. 动物准备
<ol><li>使用 Zoletil 50 （50mg/kg， IM） 和盐酸甲苯噻嗪 （40mg/kg， IP） 诱导并维持大鼠的深度麻醉。确保在整个手术过程中使用连接到加热垫的直肠探针将体温保持在 37 ± 0.5 °C。在大鼠的眼睛上涂抹兽药膏，以防止它们干燥。</li>
	<li>剃掉大鼠头部和颈部的毛发。使用脱毛膏去除头部和颈部的皮毛，然后用生理盐水洗掉乳霜。使用无菌棉球涂抹乙醇和聚维酮碘 3 次，对手术部位的皮肤进行消毒。</li>
	<li>用手术刀沿矢状缝合线的方向在大鼠头部中间切开 2 厘米，小心地去除覆盖颅骨的肌肉。</li>
	<li>使用颅骨钻头将大鼠缺血侧的颅骨变薄。在研磨过程中使用生理盐水冷却并清除碎屑。使用激光散斑对比成像 （LSCI） 记录大鼠的基线血流。</li>
	<li>使用无菌棉球将乙醇和聚维酮碘涂抹在颈部皮肤上 3 次。使用手术刀片沿颈部中线切开 2 厘米。使用牵开器横向拉回皮肤和唾液腺，露出胸锁乳突肌和颈椎肌肉。</li>
	<li>将胸锁乳突肌和颈椎肌分开，露出颈动脉区域。确定颈总动脉 （CCA）、颈内动脉 （ICA） 和颈外动脉 （ECA） 的血管解剖结构。根据血管解剖结构，将 CCA 以及 CCA 衍生的 ECA 和 ICA15 分开。 注：大鼠的解剖结构表明 CCA 区域主要被胸锁乳突肌和颈肌覆盖，其中 ICA 和 ECA 来源于 CCA。分离后露出 CCA，沿着 CCA 可以看到一个 Y 形叉子，代表 ICA 和 ECA。小心不要损伤与 CCA 平行的迷走神经。</li>
</ol>2. MCA 的闭塞
<ol><li>使用 3-0 丝线在颈外动脉 （ECA） 和颈总动脉 （CCA） 上打一个容易解开的结，以暂时阻止血流。将容器夹放在 CCA 上，距离第一个结约 0.5 厘米。<ol><li>将 5 mL 注射器的针头染成黑色。使用黑色染色的针头在 CCA 中打一个小孔，并用黑色标记针孔。</li>
	</ol></li>
	<li>通过黑色标记将单丝尼龙缝合线插入 CCA。打开血管夹，将尼龙丝引导至颈内动脉 （ICA），直到它因轻微阻力而停止。牢固地拧紧第二个结，将单丝尼龙缝合线保持在动脉内，防止从阻塞位置移位。</li>
	<li>记下此时的缺血时间，并使用 LSCI 测量缺血侧的血流值。将布比卡因滴剂滴到颈部伤口上，以镇痛和闭合伤口。取下麻醉面罩，让大鼠恢复。 注意：如果单丝尼龙缝合线难以进入 ICA，请稍微缩回并再次尝试插入。</li>
</ol>3. MCAO 的再灌注
<ol><li>缺血 90 分钟后，用异氟醚重新麻醉大鼠。使用 LSCI 测量来自缺血侧的脑血流，并确保单丝尼龙缝合线没有移动。</li>
	<li>解开颈外动脉 （ECA） 上的结，以便血液再灌注。松开颈总动脉 （CCA） 上的结，固定单丝缝合线，然后撤回缝合线。在固定单丝缝合线之前应用血管夹以防止出血。</li>
	<li>用血管夹替换第一个 CCA 结。使用镊子将血管切口侧向旋转（图 1）。用镊子夹住切口，并使用 6-0 线结扎以修复切口。取下血管夹，检查是否有渗漏，并确认完全再灌注（视频 1）。 注意：进行结扎以修复切口，以避免过度夹住血管，这可能导致 CCA 狭窄。</li>
	<li>切口结扎后使用 LSCI 记录脑皮层血流值，以确认再灌注成功。用缝合线闭合颈部切口，并给予 1,00,000 单位青霉素和 2 mL 生理盐水，以防止感染和脱水。</li>
	<li>保持温暖，直到大鼠恢复意识。术后提供软食并监测动物的生命体征。</li>
</ol>4. 神经功能与脑缺血性损伤的评估
<ol><li>一旦大鼠完全醒来，研究人员将评估它们的神经功能，而不了解动物分组。利用 Longa 评分量表（表 1）和改良的神经严重程度评分 （mNSS） 量表（表 2）来评估所有大鼠的神经功能。</li>
	<li>手术后 24 小时后，在异氟醚诱导的麻醉下（4% 异氟醚，4 L/min 氧气） 通过 颈椎脱位16 （遵循机构批准的方案）对大鼠实施安乐死。用冰水冲洗大脑以去除任何残留的血液。</li>
	<li>将脑组织分成 5 个部分并用 TTC 染色。捕获脑组织切片两侧的图像，使用 Image J 软件测量前后两侧的缺血面积，并使用以下公式计算梗死率： <img alt="Equation 1" src="/files/ftp_upload/67060/67060eq01.jpg" style="margin: auto;"> 注：S1 和 S2 分别代表脑切片近端前脑和近端脑干侧的梗死区域。使用 Swanson 公式校正每个脑切片的梗塞区域，其中 h 表示脑切片的厚度。该公式由作者先前在先前的研究17 中概述，允许更准确地计算梗死病变的大小。</li>
	<li>使用统计和绘图软件分析数据，以评估模型的成功率和稳定性。</li>
	<li>采用免疫荧光法标记大鼠的皮层神经元，进一步验证该方法引起的神经元损伤。大鼠缺血再灌注 24 小时后，使用预冷 PBS 进行心脏灌注以获得脑组织。</li>
	<li>将脑组织嵌入并冷冻成切片。使用神经元抗体，例如未成熟神经元标志物双皮质素 （DCX）18、神经元核标志物神经元核 （NeuN）18 和神经元树突状标志物微管相关蛋白-2 （MAP-2）19。使用激光共聚焦显微镜观察缺血神经元的表达。</li>
</ol>

保留 ECA 的改良 MCAO/R 模型，用于增强再灌注和稳定性

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E., Janus, J., Fern, R., Gibson, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+alternative+surgical+approach+reduces+variability+following+filament+induction+of+experimental+stroke+in+mice.">An alternative surgical approach reduces variability following filament induction of experimental stroke in mice.</a> Dis Model Mech. 10 (7), 931-938 (2017).</li></ol>

由单丝尼龙缝合线诱导的大脑中动脉闭塞 （MCAO） 模型是制备 MCAO 模型的最常用方法。这种方法在临床前研究中被广泛采用，并因其简单、无需开颅手术、手术创伤最小以及能够实现再灌注而获得许多从业者的认可。
腔内细丝 MCAO 有两种经典的手术技术：小泉法8 和 Longa 法9。这些方法之间的主要区别在于如何引入缝合线以闭塞 MCA。在 Koizu...

根据世界卫生组织的数据，在过去十年中，中风一直是全球第二大死因，发病率高、死亡率高、残疾率高 1,2。随着全球人口老龄化，预计发展中国家的中风发病率将增加，可能成为成人过早死亡和残疾的主要原因。此外，中风有发生在年轻年龄的趋势3。中风后劳动力的流失也给家庭和社会带来了沉重的负担4。因此，开发安全有效的治疗方法对中风研究构成了重大挑战。
动物模型是研究人类疾病预防和治疗的重要工具。卒中治疗策略的成功转化取决于卒中动物模型的可重复性和可靠性 5,6。大脑中动脉 （MCA） 是临床卒中的常见部位，使 MCAO 模型成为最接近人类缺血性卒中的模型。使用缝合法制备的 MCAO 模型因其无需开颅手术和易于控制缺血时间等优点而受到研究人员的青睐。它已被用于超过 40% 的神经保护实验7。然而，尽管它有许多优点，但该模型的操作细节对许多研究人员来说仍然是一个有争议的话题。
对于缝合诱导的大脑中动脉闭塞 （MCAO） 模型，通过撤回缝合线进行再灌注。目前，缝合插入主要采用两种方法：小泉的方法8 和 Longa 的方法9。在 Koizumi 的方法中，缝合线主要通过颈总动脉 （CCA） 切口进入颈内动脉 （ICA），而在 Longa 的方法中，它通过切断的颈外动脉 （ECA） 进入 ICA。在再灌注过程中，小泉法需要永久结扎 CCA 切口，并依靠 Willis 环进行再灌注10。然而，一些研究表明，在失去 CCA 供应后，仅通过对 Willis 环的补偿供应，不能仅通过补偿性供应来实现有效的再灌注。此外，Willis 环表现出很高的解剖变异性，尤其是在 C57Bl/6 小鼠中，增加了梗死变异性并降低了实验数据的可靠性。因此，这种方法越来越多地受到研究人员的质疑11。
Longa 的方法包括通过切断的 ECA 插入缝合线，然后在缝合线拔出后永久结扎颈内动脉 （ICA）。这保持了 CCA 通畅性，允许血液灌注高达基线值的 100%。然而，这种方法需要将颈外动脉和小动脉分支分开，切断它们或电凝它们，使手术具有挑战性。它还会破坏大脑的全血流结构，这与临床患者的状态不同12。重要的是，研究表明，切割或结扎 ECA 会导致控制咀嚼和吞咽的肌肉出现缺血性病变，影响动物饮食并导致大鼠术后动物死亡和严重的感觉和运动损伤13,14。
因此，迫切需要一种改进的模型制备方法来解决这些问题。本研究介绍了一种改进的 MCAO 建模方法，该方法可修复 CCA 插入切口并实现有效的再灌注。该程序简单、实用、可行，可诱导严重的神经损伤和可复制的梗死病变，为卒中研究人员提供有价值的指导。

<table><tbody><tr><td>Animal anesthesia system</td><td>Rayward Life Technology Co., Ltd</td><td>R500IE</td><td></td></tr><tr><td>Animal temperature maintainer</td><td>Rayward Life Technology Co., Ltd</td><td>69020</td><td></td></tr><tr><td>Cy3 secondary antibody</td><td>Wuhan Saiweier Biotechnology Co., Ltd</td><td>GB21303</td><td></td></tr><tr><td>DAP1 antibody</td><td>Wuhan Saiweier Biotechnology Co., Ltd</td><td>G1012</td><td></td></tr><tr><td>DCX antibody</td><td>Wuhan Saiweier Biotechnology Co., Ltd</td><td>GB13434</td><td></td></tr><tr><td>Goat serum</td><td>Beyotime Biotechnology Co., LTD</td><td>C0265</td><td></td></tr><tr><td>GraphPad Prism</td><td>GraphPad Software</td><td>GraphPad Prism 8.0</td><td></td></tr><tr><td>ImageJ</td><td>National Institutes of Health</td><td>ImageJ software</td><td></td></tr><tr><td>Isofluran</td><td>Rayward Life Technology Co., Ltd</td><td>R510-22</td><td></td></tr><tr><td>Laser speckle blood flow imaging system</td><td>Rayward Life Technology Co., Ltd</td><td>PeriCam PSI NR</td><td></td></tr><tr><td>MAP-2 antibody</td><td>Wuhan Saiweier Biotechnology Co., Ltd</td><td>GB11128</td><td></td></tr><tr><td>Miniature hand-held skull drill</td><td>Rayward Life Technology Co., Ltd</td><td>87001</td><td></td></tr><tr><td>monofilament suture</td><td>Rayward Life Technology Co., Ltd</td><td>250-280g</td><td></td></tr><tr><td>NeuN antibody</td><td>Wuhan Saiweier Biotechnology Co., Ltd</td><td>GB11138</td><td></td></tr><tr><td>OCT embedding agent</td><td>BIOSHARP</td><td>BL557A</td><td></td></tr><tr><td>Penicillin sodium</td><td>Chengdu Kelong Chemical Co., Ltd.</td><td>17121709-2</td><td></td></tr><tr><td>Quick Antigen Retrieval Solution for Frozen Sections</td><td>Beyotime Biotechnology Co., LTD</td><td>P0090</td><td></td></tr><tr><td>SD rats</td><td>SPF ( Beijing ) Biotechnology Co.,Ltd.</td><td>250-280g</td><td></td></tr><tr><td>Triton X-100</td><td>Beyotime Biotechnology Co., LTD</td><td>ST795</td><td></td></tr><tr><td>TTC</td><td>Chengdu Kelong Chemical Co., Ltd.</td><td>2019030101</td><td></td></tr></tbody></table>

a modified model preparation for middle cerebral artery occlusion reperfusion

大脑中动脉闭塞再灌注 （MCAO/R） 模型对于理解卒中的病理机制和药物开发至关重要。然而，在常用的建模方法中，小泉法由于结扎颈总动脉 （CCA） 且无法实现足够的再灌注而经常面临审查。同样，Longa 方法因断开和结扎颈外动脉 （ECA） 而受到批评。本研究旨在介绍一种改进的模型制备方法，该方法保留了 ECA 的完整性，包括通过 CCA 插入单丝尼龙缝合线，修复结扎的 CCA 切口，并维持 CCA 的再灌注。使用激光斑点流成像确认血流再灌注。Longa 量表、改良神经严重程度评分、氯化三苯基四唑 （TTC） 染色和神经元免疫荧光标记等评估方法表明，这种方法可以诱导稳定的缺血性神经损伤。这种改进的 MCAO/R 模型方案简单稳定，为脑缺血领域的从业者提供了有价值的指导。

激光斑点血流成像显示，在单丝尼龙缝合线闭塞前，大脑中动脉 （MCA） 区血流充沛，记录大鼠基线血流值。MCA 闭塞后，大脑缺血侧的血流值迅速下降。在撤回缝线之前，重新检查缺血侧的血流值，以确认缝线是否阻塞了 MCA。结果表明血流仅发生轻微变化。撤回缝合线后，血流灌注迅速恢复（图2）。 视频 1 还显示了颈总动脉的强劲充盈脉搏，表明该方法可以?...

作者聚焦：脑缺血建模的新方法 – 增强再灌注和简化手术

该协议描述了准备通过颈总动脉进行大脑中动脉闭塞再灌注的过程。

大脑中动脉闭塞再灌注的改良模型制备

The middle cerebral artery occlusion reperfusion (MCAO/R) model is crucial for understanding the pathological mechanisms of stroke and for drug ...

a-modified-model-preparation-for-middle-cerebral-artery-occlusion

Research

JoVE Journal

Medicine

A Modified Model Preparation for Middle Cerebral Artery Occlusion Reperfusion

840 次观看.  Foshan University. 我们的研究范围是脑缺血的建模程序。我们尝试从颈总动脉引入丝线并用细丝线结扎切口，维持颈总动脉和颈外动脉的再灌注。小泉法被认为再灌注不足，但 Longa 法被认为会影响主肌的血液供应。操作复杂，对初学者不友好。我们的协议解决了上述问题，并提供了一种替代方法。我们的方案使更多从业者更容易掌握脑缺血建模，并快速发现潜?...

视频: 作者聚焦：脑缺血建模的新方法 – 增强再灌注和简化手术

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

科研

神经科学

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

医学

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

颅内动脉闭塞允许经普通颈动脉修复再灌注的研究

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

脑缺血再灌注的双血管闭塞小鼠模型

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

模拟小鼠中风:通过颈外动脉的短暂性脑中动脉闭塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

建立经颅大脑中动脉闭塞小鼠模型

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. The filament MCAO model typically exhibits a massive cerebral infarction in C57Bl/6 mice that sometimes includes brain tissue in the territory supplied by the posterior cerebral artery, which is largely due to a high incidence of posterior communicating artery atresia. This phenomenon is considered a major contributor to the high mortality rate observed in C57Bl/6 mice during long-term stroke recovery after filament MCAO. Thus, many chronic stroke studies exploit distal MCAO models. However, these models usually produce infarction only in the cortex area, and consequently, the assessment of post-stroke neurologic deficits could be a challenge. This study has established a modified transcranial MCAO model in which the MCA at the trunk is partially occluded either permanently or transiently via a small cranial window. Since the occlusion location is relatively proximal to the origin of the MCA, this model generates brain damage in both the cortex and striatum. Extensive characterization of this model has demonstrated an excellent long-term survival rate, even in aged mice, as well as readily detectable neurologic deficits. Therefore, the MCAO mouse model described here represents a valuable tool for experimental stroke research.

作者聚焦:中风研究的创新技术和未来方向 

This protocol demonstrates a unique mouse stroke model with a medium-sized infarct and an excellent survival rate. This model allows preclinical stroke researchers to extend the ischemia duration, use aged mice, and assess long-term functional outcomes.

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...

大脑中动脉闭塞再灌注的改良模型制备

副本

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此视频中的章节

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

对大脑中动脉闭塞再灌注损伤诱发中风小鼠的手术方法

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

颅内动脉闭塞允许经普通颈动脉修复再灌注的研究

脑缺血再灌注的双血管闭塞小鼠模型

模拟小鼠中风:通过颈外动脉的短暂性脑中动脉闭塞

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局