Name: pre-chiasmatic, single injection of autologous blood to induce experimental subarachnoid hemorrhage in a rat model
Uploaded: 2021-06-18T14:00:00
Description: Watch this Scientific Journal Video about Pre-Chiasmatic, Single Injection of Autologous Blood to Induce Experimental Subarachnoid Hemorrhage in a Rat Model at JoVE.com

这项工作得到了灵北基金会和灵北卓越奖（编号R59-A5404）的支持。资助者在手稿的任何部分都没有角色。

该程序是根据欧盟关于保护用于科学目的的动物的指令2010/63 / EU完成的，并由丹麦动物实验检查局批准（许可证号2016-15-0201-00940）。尽可能使用无菌技术进行手术，包括无菌器械、手套、导管和缝合线。该研究使用体重230-350g的雄性和雌性Sprague-Dawley大鼠，组饲养12小时光照/黑暗循环，恒温22°C（±2°C），湿度为55%（±10%）。为动物提供标准的食物和随意的水。手术后将动物饲养在单笼中，但当ICP?...

<ol>
	<li>van Gijn, J., Kerr, R. S., Rinkel, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+haemorrhage.">Subarachnoid haemorrhage.</a> Lancet. 369 (9558), 306-318 (2007).</li><li>de Rooij, N. K., Linn, F. H. H., vander Plas, J. A., Algra, A., Rinkel, G. J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+of+subarachnoid+haemorrhage:+a+systematic+review+with+emphasis+on+region,+age,+gender+and+time+trends.">Incidence of subarachnoid haemorrhage: a systematic review with emphasis on region, age, gender and time trends.</a> Journal of Neurology, Neurosurgery, and Psychiatry. 78 (12), 1365-1372 (2007).</li><li>Feigin, V. L., Lawes, C. M., Bennett, D. A., Barker-Collo, S. L., Parag, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Worldwide+stroke+incidence+and+early+case+fatality+reported+in+56+population-based+studies:+a+systematic+review.">Worldwide stroke incidence and early case fatality reported in 56 population-based studies: a systematic review.</a> The Lancet, Neurology. 8 (4), 355-369 (2009).</li><li>Maher, M., Schweizer, T. A., Macdonald, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+spontaneous+subarachnoid+hemorrhage:+guidelines+and+gaps.">Treatment of spontaneous subarachnoid hemorrhage: guidelines and gaps.</a> Stroke. 51 (4), 1326-1332 (2020).</li><li>Pickard, J. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+oral+nimodipine+on+cerebral+infarction+and+outcome+after+subarachnoid+haemorrhage:+British+aneurysm+nimodipine+trial.">Effect of oral nimodipine on cerebral infarction and outcome after subarachnoid haemorrhage: British aneurysm nimodipine trial.</a> British Medical Journal (Clinical Research ed.). 298 (6674), 636-642 (1989).</li><li>Daou, B. J., Koduri, S., Thompson, B. G., Chaudhary, N., Pandey, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+and+experimental+aspects+of+aneurysmal+subarachnoid+hemorrhage.">Clinical and experimental aspects of aneurysmal subarachnoid hemorrhage.</a> CNS Neuroscience and Therapeutics. 25 (10), 1096-1112 (2019).</li><li>Fujii, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+brain+injury,+an+evolving+frontier+in+subarachnoid+hemorrhage+research.">Early brain injury, an evolving frontier in subarachnoid hemorrhage research.</a> Translational Stroke Research. 4 (4), 432-446 (2013).</li><li>Roos, Y. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+and+outcome+in+patients+with+aneurysmal+subarachnoid+haemorrhage:+A+prospective+hospital+based+cohort+study+in+the+Netherlands.">Complications and outcome in patients with aneurysmal subarachnoid haemorrhage: A prospective hospital based cohort study in the Netherlands.</a> Journal of Neurology, Neurosurgery, and Psychiatry. 68 (3), 337-341 (2000).</li><li>Vergouwen, M. D. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definition+of+delayed+cerebral+ischemia+after+aneurysmal+subarachnoid+hemorrhage+as+an+outcome+event+in+clinical+trials+and+observational+studies:+proposal+of+a+multidisciplinary+research+group.">Definition of delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage as an outcome event in clinical trials and observational studies: proposal of a multidisciplinary research group.</a> Stroke. 41 (10), 2391-2395 (2010).</li><li>Brown, R. J., Kumar, A., Dhar, R., Sampson, T. R., Diringer, M. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+delayed+infarcts+and+angiographic+vasospasm+after+aneurysmal+subarachnoid+hemorrhage.">The relationship between delayed infarcts and angiographic vasospasm after aneurysmal subarachnoid hemorrhage.</a> Neurosurgery. 72 (5), 702-707 (2013).</li><li>Dhar, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+angiographic+vasospasm+and+regional+hypoperfusion+in+aneurysmal+subarachnoid+hemorrhage.">Relationship between angiographic vasospasm and regional hypoperfusion in aneurysmal subarachnoid hemorrhage.</a> Stroke. 43 (7), 1788-1794 (2012).</li><li>Macdonald, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+neurological+deterioration+after+subarachnoid+haemorrhage.">Delayed neurological deterioration after subarachnoid haemorrhage.</a> Nature Reviews. Neurology. 10 (1), 44-58 (2014).</li><li>Marbacher, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+review+of+in+vivo+animal+models+of+subarachnoid+hemorrhage:+species,+standard+parameters,+and+outcomes.">Systematic review of in vivo animal models of subarachnoid hemorrhage: species, standard parameters, and outcomes.</a> Translational Stroke Research. 10 (3), 250-258 (2019).</li><li>Prunell, G. F., Mathiesen, T., Svendgaard, N. -. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+experimental+model+in+rats+for+study+of+the+pathophysiology+of+subarachnoid+hemorrhage.">A new experimental model in rats for study of the pathophysiology of subarachnoid hemorrhage.</a> Neuroreport. 13 (18), 2553-2556 (2002).</li><li>Prunell, G. F., Mathiesen, T., Diemer, N. H., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+subarachnoid+blood+volume,+mortality+rate,+neuronal+death,+cerebral+blood+flow,+and+perfusion+pressure+in+three+different+rat+models.">Experimental subarachnoid hemorrhage: subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models.</a> Neurosurgery. 52 (1), 165-176 (2003).</li><li>Prunell, G. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Subarachnoid+Hemorrhage:+Cerebral+blood+flow+and+brain+metabolism+during+the+acute+phase+in+three+different+models+in+the+rat.">Experimental Subarachnoid Hemorrhage: Cerebral blood flow and brain metabolism during the acute phase in three different models in the rat.</a> Neurosurgery. 54 (2), 426-437 (2004).</li><li>Velthuis, B. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage:+Aneurysm+detection+and+preoperative+evaluation+with+CT+angiography.">Subarachnoid hemorrhage: Aneurysm detection and preoperative evaluation with CT angiography.</a> Radiology. 208 (2), 423-430 (1998).</li><li>Leclerc, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+pathophysiology+in+humans+and+rodent+models+of+subarachnoid+hemorrhage.">A comparison of pathophysiology in humans and rodent models of subarachnoid hemorrhage.</a> Frontiers in Molecular Neuroscience. 11, 71 (2018).</li><li>Prunell, G. F., Svendgaard, N. A., Alkass, K., Mathiesen, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation+in+the+brain+after+experimental+subarachnoid+hemorrhage.">Inflammation in the brain after experimental subarachnoid hemorrhage.</a> Neurosurgery. 56 (5), 1082-1091 (2005).</li><li>Prunell, G. F., Svendgaard, N. A., Alkass, K., Mathiesen, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+cell+death+related+to+acute+cerebral+blood+flow+changes+following+subarachnoid+hemorrhage+in+the+rat+brain.">Delayed cell death related to acute cerebral blood flow changes following subarachnoid hemorrhage in the rat brain.</a> Journal of Neurosurgery. 102 (6), 1046-1054 (2005).</li><li>Spray, S., Haanes, K. A., Edvinsson, L., Johansson, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subacute+phase+of+subarachnoid+haemorrhage+in+female+rats:+increased+intracranial+pressure,+vascular+changes+and+impaired+sensorimotor+function.">Subacute phase of subarachnoid haemorrhage in female rats: increased intracranial pressure, vascular changes and impaired sensorimotor function.</a> Microvascular Research. 135, 104127 (2020).</li><li>Ansar, S., Vikman, P., Nielsen, M., Edvinsson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebrovascular+ETB,+5-HT1B,+and+AT1+receptor+upregulation+correlates+with+reduction+in+regional+CBF+after+subarachnoid+hemorrhage.">Cerebrovascular ETB, 5-HT1B, and AT1 receptor upregulation correlates with reduction in regional CBF after subarachnoid hemorrhage.</a> American Journal of Physiology - Heart and Circulatory Physiology. 293 (6), 3750-3758 (2007).</li><li>Hansen-Schwartz, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage+enhances+endothelin+receptor+expression+and+function+in+rat+cerebral+arteries.">Subarachnoid hemorrhage enhances endothelin receptor expression and function in rat cerebral arteries.</a> Neurosurgery. 52 (5), 1188-1194 (2003).</li><li>Hayman, E. G., Wessell, A., Gerzanich, V., Sheth, K. N., Simard, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+global+cerebral+edema+formation+in+aneurysmal+subarachnoid+hemorrhage.">Mechanisms of global cerebral edema formation in aneurysmal subarachnoid hemorrhage.</a> Neurocritical Care. 26 (2), 301-310 (2017).</li><li>Miyata, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vasa+vasorum+formation+is+associated+with+rupture+of+intracranial+aneurysms.">Vasa vasorum formation is associated with rupture of intracranial aneurysms.</a> Journal of Neurosurgery. , 1-11 (2019).</li><li>Tada, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+hypertension+in+the+rupture+of+intracranial+aneurysms.">Roles of hypertension in the rupture of intracranial aneurysms.</a> Stroke. 45 (2), 579-586 (2014).</li><li>Nuki, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elastase-induced+intracranial+aneurysms+in+hypertensive+mice.">Elastase-induced intracranial aneurysms in hypertensive mice.</a> Hypertension. 54 (6), 1337-1344 (1979).</li><li>Marbacher, S., Wanderer, S., Strange, F., Gr&#252;ter, B. E., Fandino, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saccular+aneurysm+models+featuring+growth+and+rupture:+A+systematic+review.">Saccular aneurysm models featuring growth and rupture: A systematic review.</a> Brain Sciences. 10 (2), 101 (2020).</li><li>Altay, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+on+brain+inflammation.">Isoflurane on brain inflammation.</a> Neurobiology of Disease. 62, 365-371 (2014).</li><li>Hockel, K., Trabold, R., Sch&#246;ller, K., T&#246;r&#246;k, E., Plesnila, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+anesthesia+on+pathophysiology+and+mortality+following+subarachnoid+hemorrhage+in+rats.">Impact of anesthesia on pathophysiology and mortality following subarachnoid hemorrhage in rats.</a> Experimental and Translational Stroke Medicine. 4 (1), 5 (2012).</li><li>Kamp, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Systematic+and+meta-analysis+of+mortality+in+experimental+mouse+models+analyzing+delayed+cerebral+ischemia+after+subarachnoid+hemorrhage.">A Systematic and meta-analysis of mortality in experimental mouse models analyzing delayed cerebral ischemia after subarachnoid hemorrhage.</a> Translational Stroke Research. 8 (3), 206-219 (2017).</li><li>Povlsen, G. K., Johansson, S. E., Larsen, C. C., Samraj, A. K., Edvinsson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+events+triggering+delayed+vasoconstrictor+receptor+upregulation+and+cerebral+ischemia+after+subarachnoid+hemorrhage.">Early events triggering delayed vasoconstrictor receptor upregulation and cerebral ischemia after subarachnoid hemorrhage.</a> BMC Neuroscience. 14, 34 (2013).</li></ol>

SAH的视交叉前单次注射模型模拟了人类SAH的几个重要元素，包括ICP峰值，CBF降低，短暂性全面缺血，神经炎症标志物上调和CVS14，15，16，18，19，20。ICP探针也用作鞘内给药的端口（图5）。此外，研究表明，该模型在雄性和雌?...

非创伤性蛛网膜下腔出血（SAH）是卒中的一种形式，约占所有病例的5%。非创伤性 SAH 最常见的原因是动脉瘤 （aSAH） 突然破裂，占 SAH 的 85%。其他原因包括动脉静脉畸形破裂、凝血病和中脑周围出血的静脉破裂1。发病率为 9/100，000 人年，死亡率约为三分之一，另外三分之一需要在 SAH2，3 后维持日常生活。
初步稳定和确诊后，治疗取决于出血的严重程度。病情最严重的患者将在心室中插入心室外引流管以降低脑内压（ICP），并被收入神经重症监护室，在那里他们受到密切监测。患者将接受血管造影以识别（可能的）动脉瘤，然后将动脉瘤盘绕或夹住以防止再出血4.尽管进行了大量药物治疗试验，但只有尼莫地平（一种钙通道拮抗剂）已被证明可以改善结局5。目前正在进行多项临床试验。请参阅Daou及其同事的评论，以获取广泛的列表6。
动脉瘤的破裂被描述为有史以来最严重的头痛或霹雳性头痛的突然发作。破裂导致ICP急剧上升，随后脑血流量（CBF）减少。这种减少导致大脑的整体缺血，这可能导致意识丧失。这种更机械的途径，以及血液中外渗元素的启动分解，引起细胞因子释放和先天免疫系统的激活，导致无菌性神经炎症。此外，经常观察到血脑屏障的破坏，导致脑水肿和离子稳态紊乱。所有这些变化以及更多被称为早期脑损伤（EBI）的变化发生在头几天内，并导致神经元丢失和细胞凋亡7。
大约 1/3 的 aSAH 患者在第 4-14 天之间会出现迟发性脑缺血 （DCI）8。DCI被定义为局灶性神经损伤的首次出现，或者在格拉斯哥昏迷量表上至少下降两个点，持续至少1小时，当排除其他原因，包括癫痫发作和再出血时。DCI 与 aSAH9 后死亡风险增加和功能结局降低有关。脑血管痉挛（CVS）是脑动脉的狭窄，几十年来一直已知与DCI有关，以前被认为是DCI的唯一原因。此后已经表明，CVS可以在没有DCI发展的情况下发生，并且已经确定了更多因素，包括微血管血栓形成和收缩，皮质扩散抑制和EBI的炎症反应10，11，12。
由于EBI和DCI对疾病进程和患者结果的巨大影响，动物模型需要尽可能模仿这些，同时仍然可重复。研究人员在从小鼠到非人类灵长类动物的各种动物中采用了各种不同的模型来尝试模拟aSAH。Sprague-Dawley和Wistar野生型大鼠是目前最常用的实验动物，最常见的模型是血管内穿孔模型，cisterna-magna双注射模型，最后是视交叉前单次注射模型，本文将在本文中描述13。
前交叉单次注射模型最初由Prunell及其同事开发，以应对其他实验模型的一些缺点14。掌握手术后，具有高度可重复性，并最大限度地减少了动物之间的差异。该模型在多个点上模拟了人类的 SAH，包括注射血液后 ICP 突然升高，导致由于 CBF15，16 下降而导致短暂性整体缺血。它影响前循环，这是人类大多数aSAH发生的地方17。死亡率在 10%-33% 之间，具体取决于研究和注射的血液量14，18。延迟性细胞死亡和神经炎症可以在第2天和第7天检测到，从而为研究EBI和DCI19，20的后果提供变量。
该研究提供了大鼠交叉前单次注射模型的更新描述，以及如何利用ICP探针作为鞘内给药的端口的描述。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>16 G peripheral vein catheter</td>
			<td>BD Venflon</td>
			<td>393229</td>
			<td>Needle shortened, distal 1 cm curved. Wings removed</td>
		</tr>
		<tr>
			<td>Anesthesia bell/ chamber</td>
			<td>Unknown</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Blood gas analyzer</td>
			<td>Radiometer</td>
			<td>ABL80</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood pressure (BP) monitor</td>
			<td>Adinstruments</td>
			<td>ML117</td>
			<td>Connects to Powerlab</td>
		</tr>
		<tr>
			<td>Curved forceps, 12 cm x 3</td>
			<td>F.S.T</td>
			<td>11001-12</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>Cylindrical pillow, 28 cm x 4 cm</td>
			<td>Homemade</td>
			<td></td>
			<td>Made from surgical towels</td>
		</tr>
		<tr>
			<td>Data acquisition hardware</td>
			<td>Adinstruments</td>
			<td>ML870 Powerlab</td>
			<td></td>
		</tr>
		<tr>
			<td>Data acquistion software</td>
			<td>Adinstruments</td>
			<td>LabChart 6.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>KMD</td>
			<td>1189</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill controller</td>
			<td>Silfradent</td>
			<td>300 IN</td>
			<td></td>
		</tr>
		<tr>
			<td>Flexible light</td>
			<td>Schott</td>
			<td>KL200</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Minco</td>
			<td>1135</td>
			<td></td>
		</tr>
		<tr>
			<td>Hypodermic needle, 20 G</td>
			<td>KD Medical</td>
			<td>301300</td>
			<td>Connects to stereotaxic frame</td>
		</tr>
		<tr>
			<td>ICP monitor</td>
			<td>Adinstruments</td>
			<td>ML117</td>
			<td>Connects to Powerlab</td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>Ohmeda</td>
			<td>TEC3</td>
			<td></td>
		</tr>
		<tr>
			<td>Laptop</td>
			<td>Lenovo</td>
			<td>T410</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser doppler monitor</td>
			<td>Adinstruments</td>
			<td>ML191</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser doppler probe</td>
			<td>Oxford Optronics</td>
			<td>MSF100XP</td>
			<td>Connects to laser doppler monitor</td>
		</tr>
		<tr>
			<td>Needle holder, 13 cm</td>
			<td>F.S.T</td>
			<td>12001-13</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>Precision syringe, 0.025 mL</td>
			<td>Hamilton</td>
			<td>547407</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic frame</td>
			<td>Kopf Instruments</td>
			<td>M900</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical microscope</td>
			<td>Carl Zeiss</td>
			<td>F170</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture needle</td>
			<td>Allgaier</td>
			<td>1245</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>Temperaure controller</td>
			<td>CWE,INC.</td>
			<td>TC-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Transducer x 2</td>
			<td>Adinstruments</td>
			<td>MLT0699</td>
			<td>Connects to BP and ICP monitor</td>
		</tr>
		<tr>
			<td>Ventilator</td>
			<td>Ugo Basile</td>
			<td>7025</td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary clipper</td>
			<td>Aesculap</td>
			<td>GT421</td>
			<td></td>
		</tr>
		<tr>
			<td>3-pronged Blair retractor, 13.5 cm</td>
			<td>Agnthos</td>
			<td>17022--13</td>
			<td></td>
		</tr>
		<tr>
			<td>Blunt Alm retractor</td>
			<td>F.S.T</td>
			<td>17008-07</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved forceps, 12 cm x 2</td>
			<td>F.S.T</td>
			<td>11001-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder, 13 cm</td>
			<td>F.S.T</td>
			<td>12001-13</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Dumont forceps, 11 cm</td>
			<td>F.S.T</td>
			<td>11252-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Halsted-Mosquito hemostat x 2</td>
			<td>F.S.T</td>
			<td>13008-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Iris scissor, 9 cm</td>
			<td>F.S.T</td>
			<td>14090-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Vannas scissor, 10.5 cm</td>
			<td>F.S.T</td>
			<td>15018-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorpable swabs</td>
			<td>Kettenbach</td>
			<td>31603</td>
			<td></td>
		</tr>
		<tr>
			<td>Black silk thread, 4-0, 5 x 15 cm</td>
			<td>V&#246;mel</td>
			<td>14757</td>
			<td></td>
		</tr>
		<tr>
			<td>Bone wax</td>
			<td>Aesculap</td>
			<td>1029754</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbomer eye gel 2 mg/g</td>
			<td>Paranova</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab</td>
			<td>Heinz Herenz</td>
			<td>WA-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton tipped applicator x 4</td>
			<td>Selefa</td>
			<td>120788</td>
			<td></td>
		</tr>
		<tr>
			<td>Hypodermic needle, 23 G x2</td>
			<td>KD Medical</td>
			<td>900284</td>
			<td>Connects to stopcock. Remove distal end</td>
		</tr>
		<tr>
			<td>Hypodermic needle, 23 G x3</td>
			<td>KD Medical</td>
			<td>900284</td>
			<td>Remove distal end. 2 connects to stopcock, 1 to syringe</td>
		</tr>
		<tr>
			<td>ICP probe:</td>
			<td>Homemade</td>
			<td></td>
			<td>Made of the following:</td>
		</tr>
		<tr>
			<td>Polythene tubing, 20 mm</td>
			<td>Smiths medical</td>
			<td>800/100/200</td>
			<td>Inner diameter (ID): 0.58 mm, Outer diameter (OD): 0.96 mm.</td>
		</tr>
		<tr>
			<td>Silicone tubing, 10 mm</td>
			<td>Fisher</td>
			<td>15202710</td>
			<td>ID: 0.76 mm, OD: 2.4 mm.</td>
		</tr>
		<tr>
			<td>Silicone tubing, 2 mm</td>
			<td>Fisher</td>
			<td>11716513</td>
			<td>ID: 1.0 mm, OD: 3.0 mm.</td>
		</tr>
		<tr>
			<td>Micro hematocrit tubes</td>
			<td>Brand</td>
			<td>7493 11</td>
			<td></td>
		</tr>
		<tr>
			<td>OP-towel, 45 cm x75 cm</td>
			<td>M&#246;lnlycke</td>
			<td>800430</td>
			<td></td>
		</tr>
		<tr>
			<td>PinPort adapter, 22 G</td>
			<td>Instech</td>
			<td>PNP3F22</td>
			<td></td>
		</tr>
		<tr>
			<td>PinPort injector</td>
			<td>Instech</td>
			<td>PNP3M</td>
			<td></td>
		</tr>
		<tr>
			<td>Polythene tubing, 2 x 20 cm</td>
			<td>Smiths medical</td>
			<td>800/100/200</td>
			<td>Connects to syringe. ID: 0.58 mm, OD: 0.96 mm.</td>
		</tr>
		<tr>
			<td>Rubberband</td>
			<td>Unknown</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel, 10 blade</td>
			<td>Kiato</td>
			<td>23110</td>
			<td></td>
		</tr>
		<tr>
			<td>Spinalneedle, 25 G x 3.5&#39;&#39;</td>
			<td>Braun</td>
			<td>5405905-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Stopcock system, Discofix x 2</td>
			<td>Braun</td>
			<td>16494C</td>
			<td>Connects to transducer</td>
		</tr>
		<tr>
			<td>Suture, 4-0, monofil, non-resorbable x 3</td>
			<td>Ethicon</td>
			<td>EH7145H</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, 1 mL</td>
			<td>BD Plastipak</td>
			<td>1710023</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, luer-lock, 10 mL x 4</td>
			<td>BD Plastipak</td>
			<td>305959</td>
			<td>Connects to transducer</td>
		</tr>
		<tr>
			<td>Tissue adhesive glue</td>
			<td>3M</td>
			<td>1469SB</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5% Chlorhexidine spirit</td>
			<td>Faaborg Pharma</td>
			<td>210918</td>
			<td></td>
		</tr>
		<tr>
			<td>Carprofen 50 mg/mL</td>
			<td>ScanVet</td>
			<td>43715</td>
			<td>Diluted 1:10</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Baxter</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isotonic saline</td>
			<td>Amgros</td>
			<td>16404</td>
			<td></td>
		</tr>
		<tr>
			<td>Lidocaine-Adrenaline 10 mg/5 &#181;g/mL</td>
			<td>Amgros</td>
			<td>16318</td>
			<td></td>
		</tr>
	</tbody>
</table>

pre-chiasmatic, single injection of autologous blood to induce experimental subarachnoid hemorrhage in a rat model

尽管过去几十年在治疗方面取得了进展，但蛛网膜下腔出血（SAH）仍然具有很高的发病率和死亡率负担，主要困扰着相当年轻的人群。已经开发了几种SAH动物模型来研究SAH背后的病理生理机制并测试药物干预。本文介绍的大鼠的视交叉前单次注射模型是具有预定血容量的SAH的实验模型。简而言之，对动物进行麻醉、插管并保持在机械通气下。温度通过加热垫调节。在尾动脉中放置导管，可以连续测量血压和进行血液采样。切开寰枕膜，并在大池中放置用于压力记录的导管，以测量脑内压。该导管也可用于鞘内治疗干预。将大鼠置于立体定位框架中，在前膛前钻一个毛刺孔，并通过毛刺孔插入导管并放置在视交叉前方。从尾导管抽出自体血 （0.3 mL） 并手动注射。这导致脑内压升高和脑血流量减少。将动物保持镇静30分钟，并给予皮下盐水和镇痛药。动物被拔管并返回笼子。前交叉模型具有很高的重现率，并且由于预先确定的血容量，动物之间的差异有限。它模仿人类的SAH，使其成为SAH研究的相关模型。

与男性相比，女性患 aSAH 的风险更高。尽管如此，雄性啮齿动物主要用于实验，因为雌性发情周期的异质性可能存在偏差。这里介绍的代表性结果来自最近比较雌性和雄性大鼠的出版物，证实该模型在雌性动物中产生的结果与雄性21相似。该研究包括34只雌性Sprague-Dawley大鼠（18只SAH和16只shams）。Shams没有将脊髓针下降到视交叉或注射血液。所有其他程序均在与SAH相同的Shams上进行...

Watch this Scientific Journal Video about Pre-Chiasmatic, Single Injection of Autologous Blood to Induce Experimental Subarachnoid Hemorrhage in a Rat Model at JoVE.com

Pre-Chiasmatic, Single Injection of Autologous Blood to Induce Experimental Subarachnoid Hemorrhage in a Rat Model

蛛网膜下腔出血继续给人类带来很高的死亡率和发病率负担。为了便于进一步研究病情及其病理生理学，提出了一种视交叉前的单次注射模型。

视交叉前，单次注射自体血以诱导大鼠模型中实验性蛛网膜下腔出血

Despite advances in treatment over the last decades, subarachnoid hemorrhage (SAH) continues to carry a high burden of morbidity and mortality, largely ...

这种蛛网膜下腔出血模型有助于我们更好地了解病理生理机制及其后果。这将有望产生治疗方法，减轻患病患者的疾病负担。该技术的主要优点是其高度可重复性，因为注射了固定量的血液。此外，它在关键参数上类似于人类蛛网膜下腔出血。要制作ICP探头，请燃烧20毫米聚乙烯管的一端，制作圆形板并保持腔开放。然后将 1 毫米硅胶管环滑过聚乙烯管，然后将 10 毫米硅胶管连接到聚乙烯管的末端。接下来，麻醉、插管和导管插入 Sprague Dawley 大鼠，然后将其置于立体定位框架中。进行局部麻醉后，从中线的针刺开始在尾部做一个8毫米的皮肤切口。在解剖显微镜下，钝化地分层解剖所有肌肉以识别寰枕膜。然后使用手臂牵开器来约束颈部肌肉组织，如果需要，将牵开器放在尾部。检查无菌ICP探头是否连接到ICP探头后，用生理盐水冲洗ICP探头，确保探头中没有气泡。使用 23 号针切开寰枕膜，打一个孔以轻轻地哄骗 ICP 探头穿过膜。轻轻拉动探头，确保其显示0至5毫米汞柱之间的脉动曲线。如果没有，请取下探头，重新检查与探头的连接，并确认流经管腔的流动。在寰枕膜包裹探针的区域，涂上两滴组织胶。然后将一毫米的硅胶管向前移向膜，然后再涂上额外的胶水，以尽量减少ICP探头移位的风险。粘合 ICP 探头后，取下牵开器。然后，使用不可吸收的单丝4-0缝合线，在切口的尾端放置一条水平床垫缝合线，在头端放置一条简单的中断缝合线。要放置激光多普勒探头，请在中线尾部做一个 15 毫米的切口，从眼睛前方开始。用镊子去除结缔组织和肌肉后，使用无菌棉签的末端作为胭脂来识别前膛和冠状缝合线。放置武装牵开器后，将 25 号脊髓针准确地放在前膛上的立体定位框架中并注意位置。接下来，从前膛上取下针头。将框架向前移动 6.5 毫米。然后更换中线的针，以标记钻孔的侧面。钻孔直到在骨骼下方识别出硬脑膜。使用直镊子，轻轻去除骨头碎片。然后用骨蜡填充腔体。接下来，在前膛右侧三到四毫米处，就在激光多普勒的冠状缝合线的正前方，钻另一个孔。注意不要穿透硬脑膜。寻找激光多普勒可以测量血流的血管。放置激光多普勒并检查值。最小值为 100 个助焊剂单位。如果取下显微镜后该值仍然可接受，请添加一滴胶水以固定探针。重新检查以确认该值是否高于 80 个通量单位。为了诱发蛛网膜下腔出血，将针头轻轻地插入半球之间中线的颅骨，直到由于颅底而感到阻力。将针头缩回一毫米，以确保正确放置在视交叉前方。为确保注射血液时获得最均匀的结果，请将针头顺时针旋转90度，使针尖指向右侧。然后取下细高跟鞋。平衡 15 分钟后，调整麻醉水平以获得 80 至 100 毫米汞柱范围内的平均动脉血压。然后进行血气分析，以确认pH值，二氧化碳分压和氧气分压在生理参数范围内。接下来，使用带有钝23号针头的一毫升注射器，从尾导管中抽出500微升血液。为避免注入空气，请用血液填充脊髓针室的死腔。将注射器中的血液体积调节到300微升后，将注射器连接到脊髓针。然后，牢牢抓住注射器，手动注射血液以超过平均动脉压。观察脑内压急剧上升，同时脑血流量急剧下降。诱导蛛网膜下腔出血30分钟后，取下针头和激光多普勒探头。并用骨蜡填充腔。使用不可吸收的单丝4-0缝合线，用两条水平床垫缝合线关闭切口。要使用ICP探头注射到大型水池中，请取下硅胶管并将针端口适配器插入聚乙烯管。为了在动物清醒时进行鞘内给药，请将针口注射器放在精密注射器上。然后通过针端口适配器进行治疗。如果不必进行鞘内给药，请切断简单的中断缝合线。然后，用剪刀尽可能缩短ICP探头，并粘合末端，以防止脑脊液泄漏。用不可吸收的单丝4-0缝合线关闭切口。在使用旋转杆试验评估粗大感觉运动功能时，与假动物相比，实验诱导的蛛网膜下腔出血大鼠在手术后24和48小时的表现明显更差。与假对照相比，实验大鼠基底动脉和大脑中动脉蛛网膜下腔出血两天后，对内皮素-1（内皮素-1受体家族的激动剂）的敏感性显着增加。实验大鼠的两条动脉对5-羧酰胺色胺的敏感性也表现出相似的增加，5-羧色胺受体家族的激动剂。手术后，可以进行神经行为测试以及组织分析，包括免疫组织化学，PCR，蛋白质印迹分析仪，以分析后果和机制。

Research

JoVE Journal

Neuroscience

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

自体血液的注入模型小鼠自发性脑出血

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model.  While ICH
accounts for 10-15% ...

科研

神经科学

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

在实验视神经脱髓鞘模型大鼠视觉诱发电位记录

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines

由亲炎细胞因子的内脑注射诱导的广度脑皮质脱叶的鼠模型

Multiple sclerosis (MS) is the most common immune-mediated disease of the central nervous system (CNS) and progressively leads to physical disability and ...

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

在鼠模型中使用聚焦超声波打开位置特定血脑屏障开口的台面方法

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic ...

Double Direct Injection of Blood into the Cisterna Magna as a Model of Subarachnoid Hemorrhage

双直接注射血液到西斯特纳麦格作为苏巴拉奇内德出血的模型

Among strokes, subarachnoid hemorrhage (SAH) consecutive to the rupture of a cerebral arterial aneurysm represents 5-9% but is responsible for about 30% ...

Massive Pontine Hemorrhage by Dual Injection of Autologous Blood

大量庞廷出血通过双注射自体血

We provide a protocol to establish a massive pontine hemorrhage model in a rat. Rats weighing about 250 grams were used in this study. One hundred ...

A Rat Model of EcoHIV Brain Infection

生态HIV脑感染的老鼠模型

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Analysis of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage with High Frequency Transcranial Duplex Ultrasound

高频颅内双体超声波的苏巴拉赫诺德出血的穆林模型脑血管痉挛分析

Cerebral vasospasm that occurs in the weeks after subarachnoid hemorrhage, a type of hemorrhagic stroke, contributes to delayed cerebral ischemia. A ...

Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging (MRI)

Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging MRI

蛛网膜下腔出血结合磁共振成像（MRI）的血管内穿孔模型

The endovascular filament perforation model to mimic subarachnoid hemorrhage (SAH) is a commonly used model - however, the technique can cause a high ...