Intramural NIH program supported the development of this model. Publication support was provided by Harvard Apparatus Inc. and ADInstruments Inc. The work by the authors was done as part of US government&#8211;funded research; however, the opinions expressed are not necessarily those of the National Institutes of Health.

下面介绍的程序进行批准的临床中心的动物护理和使用委员会在美国国立卫生研究院的协议的一部分。 1.准备手术<ol><li>燃气导管消毒和器械。 </li></ol> 2.准备导管<ol><li>同花顺肝素甘油（250 IU肝素/ 1毫升的丙三醇）的导管，以确保通畅，避免泄漏。塞与金属塞子（20 GA 15毫米）至保留肝素甘油导管的末端。 </li></ol> 3.准备无菌工作站<ol><li>用70％的酒精喷工作站和仪表盘。使用无菌悬垂覆盖手术台和仪表盘和地点仪器的托盘上。 </li></ol> 4，大鼠准备外科手术<ol><li>地方大鼠分别在有机玻璃室，并暴露于麻醉剂;异氟烷（3-5％）的混合编氧气，直到昏迷。从腔室中取出大鼠和从颈部的前部和后部区域剃毛皮。涂抹润滑油膏眼睛。 </li></ol> 5.导管植入<ol><li>将麻醉的大鼠（由脚趾捏确认）到手术加热表中腹的位置;把它的鼻子到鼻锥为维持麻醉自发性呼吸（异氟烷，3.5％）。轻轻擦洗手术区3次交替使用聚维酮碘和70％的酒精。使使用外科剪刀的肩甲骨之间的0.5厘米中线皮肤切口。 </li><li>重新定位大鼠在仰卧位;轻轻抑制腿部用橡皮筋表中的每一侧;维持麻醉。放置在颈部2推出无菌纱布的4x4小幅hyperextend更好的曝光。使在使用手术刀锁骨水平颈部的中线的2厘米腹侧颈部皮肤切口权利。 </li></ol> 6.右键颈内静脉穿刺置管<ol><li>使用止血钳，直截了当地解剖右颈静脉，分离出唾液腺和淋巴组织可视化和隔离容器的一个5毫米部分。用4-0丝线缝合，放置在容器的两个头部和尾部端部的松动领带最大化容器的曝光。用微型手术剪做一个切口足够大以通过导管，管线与两个连字之间的容器中，并配合在容器周围的颅结扎。 </li><li>将静脉导管插入朝向与微解剖钩和镊子的辅助心脏的容器中，并推进该导管，直到所有的PU 3F段是在容器中。使用连字在头部和尾部端以固定导管的容器中。 </li></ol> 7.左颈动脉插管<ol><li>用止血钳，直言解剖肩胛舌骨肌纵向暴露左侧颈总动脉和我Alcalase碱性蛋白酶容器的5毫米部分。确保迷走神经（白色的），从动脉完全分离。小心不要切碎或破坏神经。 </li><li>用4-0丝线缝合，放置在容器的尾端的松散的领带，领带关闭容器的颅端和尾端放置缝合上述哈巴狗夹停止血流以下的切口。用微型手术剪，使一个切口，大到足以与两个连字之间的容器通过导管，管线。将朝心脏动脉导管与微观解剖钩钳的帮助。 </li><li>使用无锁的平滑针保持器除去牛头犬钳紧之前保持在容器内的导管的所述部分。推进导管有一对镊子的同时松开针保持器慢慢直到导管的整个聚氨酯2F段是在该容器中。配合宽松的尾部结扎导管周围和血管确保，但不能因此紧固密封吨，以堵塞，导管。 </li></ol> 8.使用直止血，隧道5cm的管皮下回耳后，并通过肩胛骨之间的切口。 Exteriorize导管通过管和取出管。 9.关闭腹切口三种不锈钢绕线夹，以及背侧切口用4-0丝线缝合固定导管外置到位。 10.手术后监测和护理<ol><li>以下导尿，终止麻醉，并恢复在卧位大鼠中与纤维素寝具一个笼子。观察大鼠在2小时的时间间隔为至少4小时，或直到表示疼痛的迹象，然后每天一次。得到酮洛芬[5毫克/千克，皮下注射（SQ）]立即张贴程序和每12小时，如果大鼠显示疼痛或困难的症状。根据我们的经验，没有老鼠都需要以下初始额外镇痛疼痛升注射。观测到的特性，将额外的剂量镇痛诱发考虑包括但不限于：姿势异常;增加/减少呼吸速率，unthrifty / ungroomed，感染/切口部位的炎症，眼分泌物，竖毛（“加标haircoat”），降低活性响应于听觉或触觉刺激。 注：大鼠恢复72小时即获得从前期导尿重“10 g将纳入研究。 </li></ol> 11.连接到血液动力系统<ol><li>以下恢复，报名参加该研究的健康大鼠（体重增加&gt; 10克）。放置大鼠的Lucite腔和麻醉如前所述。 </li><li>将无菌的悬垂在手术台上用无菌止血钳，微型镊子1毫升注射器用钝针连接。 注：该止血钳的尖端应覆盖有橡胶，以避免损坏导管。将麻醉大鼠的手术表中的腹侧的位置和地点鼻子到鼻锥维持麻醉（3-5％）具有自发呼吸。 </li><li>钳动脉并用一对橡胶封止血略低于金属插针的静脉导管。用一对镊子除去金属销在导管的端部，并附加一个1ml注射器填充有肝素盐水附有一钝针。反过来，取出止血，收回0.1毫升，以确保导管的专利。 </li><li>再夹紧导管，取出针，并为此附加到一个较长的导管（预冲洗用肝素盐水），在连接到一个旋转，并拴在鞋盒笼子的顶部。连接这些导管的换能器的数据采集和记录，取样，或输注。 注：所有导管保护，螺旋簧管足够长的时间，使动物的全套动作的笼子里。 </li><li>下面导管连接，终止麻醉在恢复与纤维素床上用品笼子里的胸骨位置动物。为了保持动脉导管通畅，人工冲洗和0.05 ml肝素盐水（50 IU / ml）的每1小时24小时这与动物的定期评估一致的线。 </li></ol> 12.采血，压力监测和药物管理局<ol><li>附加空注射器活塞连接到动脉导管，收回0.6毫升在线冲洗。附加样品注射器，取样品，并返回0.6毫升在线冲洗后0.7毫升肝素生理盐水冲洗。 </li><li>使用数据采集硬件和软件来测量和记录MAP和心脏率不断。 </li><li>自动给药系统（ADDS）连接到静脉导管。 注：将调节血管加压输注速率（高，低，或0剂量）的基础上检测到正常生理范围内维持压力在MAP。 </li></ol>

<ol>
	<li>Buckingham, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Indwelling+catheters+for+direct+recording+of+arterial+blood+pressure+and+intravenous+injection+of+drugs+in+the+conscious+rat.">Indwelling catheters for direct recording of arterial blood pressure and intravenous injection of drugs in the conscious rat.</a> J Pharm. Pharmacol. 28 (5), 459-461 (1976).</li><li>Buckle, J. W., Nathaniels, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proceedings:+A+dual+catheter+system+for+the+simultaneous+infusion+and+sampling+of+the+vascular+system+of+the+unrestrained+rat.">Proceedings: A dual catheter system for the simultaneous infusion and sampling of the vascular system of the unrestrained rat.</a> J. Physiol. 242 (2), 55P-56P (1974).</li><li>Solomon, S. 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=Effective+dosing+of+lipid+A+analogue+E5564+in+rats+depends+on+the+timing+of+treatment+and+the+route+of+Escherichia+coli+infection.">Effective dosing of lipid A analogue E5564 in rats depends on the timing of treatment and the route of Escherichia coli infection.</a> J. Infect. Dis. 193 (5), 634-644 (2006).</li><li>Cui, X., 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=Bacillus+anthracis+cell+wall+produces+injurious+inflammation+but+paradoxically+decreases+the+lethality+of+anthrax+lethal+toxin+in+a+rat+model.">Bacillus anthracis cell wall produces injurious inflammation but paradoxically decreases the lethality of anthrax lethal toxin in a rat model.</a> Intensive Care Med. 36 (1), 148-156 (2010).</li><li>Quezado, Z. M., Natanson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+hemodynamic+abnormalities+and+vasopressor+therapy+in+sepsis+and+septic+shock.">Systemic hemodynamic abnormalities and vasopressor therapy in sepsis and septic shock.</a> Am. J. Kidney Dis. 20 (3), 214-222 (1992).</li><li>Perdue, P. W., Kazarian, K. K., Nevola, J., Law, W. R., Williams, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+local+and+systemic+antibiotics+in+rat+fecal+peritonitis.">The use of local and systemic antibiotics in rat fecal peritonitis.</a> J. Surg. Res. 57 (3), 360-365 (1994).</li><li>Qiu, P., 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=The+individual+survival+benefits+of+tumor+necrosis+factor+soluble+receptor+and+fluid+administration+are+not+additive+in+a+rat+sepsis+model.">The individual survival benefits of tumor necrosis factor soluble receptor and fluid administration are not additive in a rat sepsis model.</a> Intensive Care Med. 37 (10), 1688-1695 (2011).</li><li>Cui, X., 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=Severity+of+sepsis+alters+the+effects+of+superoxide+anion+inhibition+in+a+rat+sepsis+model.">Severity of sepsis alters the effects of superoxide anion inhibition in a rat sepsis model.</a> J. Appl. Physiol. 97 (4), 1349-1357 (2004).</li><li>Karzai, W., 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=Protection+with+antibody+to+tumor+necrosis+factor+differs+with+similarly+lethal+Escherichia+coli+versus+Staphylococcus+aureus+pneumonia+in+rats.">Protection with antibody to tumor necrosis factor differs with similarly lethal Escherichia coli versus Staphylococcus aureus pneumonia in rats.</a> Anesthesiology. 99 (1), 81-89 (2003).</li><li>Jespersen, B., Knupp, L., Northcott, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+Arterial+and+Venous+Catheterization+for+Blood+Sampling,+Drug+Administration+and+Conscious+Blood+Pressure+and+Heart+Rate+Measurements.">Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements.</a> J. Vis. Exp. (59), e3496 (2012).</li></ol>

The authors have no potential conflicts of interest.

<html>调查治疗方法的疾病的临床模型就必须准确地执行血液动力学监测，因此需要血管通路的能力。在我们的脓毒症模型，动脉导管提供了系统性压力监测和静脉插管允许的标准临床治疗中的应用。标准治疗脓毒症，包括抗生素输液治疗细菌感染和流体和升压药的输注以保持在所需范围内的动脉压力。两个导管用于采血。在此设计中，可编程输注应用与自动触发基于实时的血管加压输注速率的泵平均动脉压是预先在临床环境，需要手动滴定由医务人员和在临床前设置使使用这种治疗方式可能用于研究大量动物的。 。e_content“&gt;一些并发症模型的发展经历了早期的这些包括动脉和来自紧缩静脉导管，对PU2和PU3成分的关系，（1-2％）的部分遮挡;颈静脉导管错位成小支，（&lt;1％）;恢复（无肺动脉血栓或其他器官损伤明显）后，不明原因猝死，（1-2％）通过保持异氟醚浓度2-3.5％与2升/分钟O 2流量解析。没有观察到出血，局部感染和自身造成损伤的其他常见的手术并发症。一旦外科熟练度达到后，每个过程应该能够在不超过15分钟完成。成功的测量作为健康恢复的速率和专利导管，可以预计，在超过一次充分训练。一个附加的动物被包含在每个研究的90％，以减少数据的丢失，需要的动物的总数和帐户失败导管可以是“只有融合“或死亡，由于并发症。 在此过程中最关键的步骤是将导管的设计和布局。该材料必须是柔软的尖端，以便不以刺穿血管壁无卷曲过于容易。导管的主体具有要更硬（比前端），以便能够提前到容器充分且在适当的位置固定。 图1显示管子的直径3被连接于动脉导管和2个不同直径的管子连接为静脉导管。这些部件滑动内彼此以减少导管直径在前端并与加入，以确保在容器或心脏导管的放置被保持锚粘在一起。 一旦回收，并连接到旋转和系绳，所述MAP和CVP波形应类似于图2中所表示的数据是为了保持动脉导管patenc监测过程中Y，每小时肝素盐水冲洗（0.05毫升），应执行。连续输液保持静脉导管通畅。在线路连接和冲洗，密切一定要注意，以确保没有气泡就行了。 ≥0.1ml的气泡可导致肺，脑或其他器官的栓塞。血液动力学波形记录和在研究过程中进行分析。当在x轴压缩时， 图3示出了在1小时的过程中的变化，MAP和CVP以及变化率和血管加压输注效果。 该程序有几个潜在的局限性。在我们的研究中，导管留专利长达11天。它是未知多久导管将保持专利超过这个期限。到所述适合的导管，有一个最小的血管的大小，这大致与啮齿动物的重量相关的，需要的动物纳入本研究为大于200克。使用更小的导管直径将导致增加的电阻，以减小流动通畅。达到中心静脉压措施，需要在研究期间的心房足以帐户大小和生长精确导管放置，并且可以是一个挑战。 一旦掌握，动脉和血管导管可提供依据各种啮齿动物模型需要血液动力学监测流体或疗法，血液采样，并输注同时最小化任何疼痛和痛苦的仪器中或处理后回收的。事实上，在与股静脉的先前JOVE出版物10描述导尿结合，我们已经成功地执行了此模型用3植入导管。

大鼠的动脉和静脉导管早已用于在实验室研究1,2导管可用于监测血液动力学参数，包括收缩压，舒张压和平均动脉压（MAP），心脏速率（HR）和中心静脉压（CVP） 3,4此外，这些导管允许的标准疗法或潜在疗法输注以及血液取样，以进一步分析的疾病或治疗的作用机制。因此，具有在小动物模型的血管通路是用于研究的措施的临床表现和治疗效果的关键。 研究重大疾病的根本原因，首先建立一个模型来模拟人的条件是很重要的。气管内接种细菌的已知量的已被成功地用于重现肺炎的发病机制，一个严重肺部感染，然后发育成败血症。3，5血流动力学监测措施，并提供标准的临床治疗关键是要模拟常规支持治疗。标准的临床治疗包括输液抗生素，它有助于清除潜在的感染和体液和升压药的管理，两种治疗采用脓毒症患者维持血压。5-7血管通路，特别是当我们研究的专利导管植入尤为重要研究性治疗败血症。 多年，来管理标准的临床治疗的啮齿动物的能力通过用用于构造可植入导管和输注技术来准确地随着时间的推移递送小体积的能力所需要的特性的材料的可用性是有限的。此外，触发输注响应于连续监测的特定的血流动力学度量的能力允许以一致和可再现的立场在使用较大的样本量典型的啮齿动物的研究ARD治疗模式8,9这些技术限制在材料和精密的 ​​硬件已经克服，但同时提供常规治疗，同时监测血流动力学参数，动脉和静脉血管通路必须是可用的。 该视频详细介绍了手术技术植入颈总动脉和颈总静脉导管在麻醉的老鼠。以下一个72小时的恢复期，动物将被重新麻醉，并连接到附连到用于将植入导管连接到血液动力学监测系统中的啮齿动物壳体的系绳和旋转设置。在研究过程中，系统允许用于流体输注根据任一提供一个给定的体积在一定的输注速率或自动化系统，可提供根据所检测到保持给定的压力范围内的平均动脉压的输注速率。补液系统采用可编程注射器泵与数据采集系统的数字输出，接口和由软件监视所述MAP进行控制。系绳和旋转设置允许在研究期间大鼠可以自由流动，同时监测压力，注入液体和升压药，而无需操纵动物进行血液取样。与每个2导管（在我们的设置）与硬件可扩展至监测仪表24的动物同时监视多达12仪表化动物允许很大的灵活性，用于研究多个分层组在每个实验中。 该导管法可以是使用类似的啮齿类动物模型需要血管通路的监测，取样和治疗长达11天的研究设施是有益的。如果研究机构在考虑这个程序与啮齿类动物为维持这些动物然后developin经验和适当的支撑结构克这些技能可以提供显著节省成本，减少每个插管大鼠总成本从约1,800至不到50美元（包括天真大鼠，导管组件和技术成本），提高调度的灵活性（天真的老鼠都是现成的与公司的交付时间表植入老鼠）。这个过程的成功依赖于执行导尿个体的技能。精通此过程中所需的培训动物已经从20至40不等的大鼠与&gt; 90％导致导管通畅率。

<table border="0" cellpadding="0" cellspacing="0" width="715">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:299px;">Name of Material/ Equipment</td>
			<td style="width:131px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">straight micro dissecting forceps</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-8102</td>
			<td>surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">delicate straight hemostatic forceps</td>
			<td>Roboz</td>
			<td style="width:119px;">RS-7114</td>
			<td>surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">delicate curved hemostatic forceps</td>
			<td>Miltex</td>
			<td style="width:119px;">&#160;7-4</td>
			<td>surgical instrument</td>
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catheterization of the carotid artery and jugular vein to perform hemodynamic measures, infusions and blood sampling in a conscious rat model

小动物模型的成功，研究重大疾病，在某种程度上，依赖模型来模拟人类状况的能力。气管内接种细菌的已知量的已被成功地用于再现肺炎，然后发育成脓毒症的发病机制。监测血液动力学参数，并提供标准的临床治疗，包括输注抗生素，​​流体和药物维持血压是至关重要的模拟在这个模型，但做常规的支持治疗有此需要动脉和静脉血管通路。该视频详细介绍了手术技术植入颈总动脉和颈总静脉导管在麻醉的老鼠。以下一个72小时的恢复期，动物将被重新麻醉，并连接到附连到所述壳体的啮齿动物相连接的植入导管的血液动力学监测系统的系绳和旋转设置。这种设置允许自由流动在研究过程中大鼠，而连续监控的压力，注入液体和药物（抗生素，升压药），并进行采血。

变化的血管压力通过流体填充导管（ 图1）传送，并转换成由血液动力波形表示的电信号（ 图2,3）。无血管通路，无法建立这些措施。波形实时流可以检测一个节拍变化击败的基础上（ 图2）和分析。压缩波形的时标允许定量发生在较长的持续时间（数分钟到数天）（ 图3，上部4的波形），它可以用在血管加压输注速率的变化相关联（ 图3，下4的波形）的变化。 <img alt="图1" src="/files/ftp_upload/51881/51881fig1.jpg" /> 图1.导管设计：在动脉和静脉导管聚氨酯，聚氨酯的部件的详细情况;。 PE，聚乙烯（段长） <a href="https://www.jove.com/files/ftp_upload/51881/51881fig1highres.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/51881/51881fig2.jpg" /> 图4典型的啮齿动物后7天导管在7秒2，实时地图波形。 <a href="https://www.jove.com/files/ftp_upload/51881/51881fig2highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/51881/51881fig3.jpg" /> 图3.时间压缩MAP的4个典型啮齿类（上4波形）后7天导尿波形超过1小时。当MAP被平均（下4波形），该值是用来触发ADDS高（&lt;90毫米汞柱） ，低（&lt;100毫米汞柱）或停止（&gt; 110）从输液泵的流量响应。 <a href="https://www.jove.com/files/ftp_upload/51881/51881fig3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model at JoVE.com

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

catheterization-carotid-artery-jugular-vein-to-perform-hemodynamic

Research

JoVE Journal

Medicine

71.3K 次观看.  National Institutes of Health. Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

视频: Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

科研

医学

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

培养一个精密的显微外科技术在大鼠颈总动脉，颈外静脉移植：介入

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

成年小鼠静脉高压型号：颈总动脉，以颈外静脉吻合。

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

血管球囊损伤和腔内管理在大鼠颈动脉

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

颈动脉介入移植中颈静脉持久性转基因表达的家兔模型

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

大鼠颈总动脉的相位对比磁共振成像

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

大鼠胡萝卜动脉压力控制段气球损伤与腹腔ventit 治疗应用

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Microsurgical Skills of Establishing Permanent Jugular Vein Cannulation in Rats for Serial Blood Sampling of Orally Administered Drug

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, which could potentially affect results. The jugular vein cannulation (JVC) in rats is a widely used model for repeated blood collection but requires adequate training of surgery skills and animal care. This article details the microsurgical procedures for establishing and maintaining a permanent JVC rat model with specific focus on the placement and sealing of the jugular cannula. The importance of monitoring physiological (e.g., body weight, food, and water intake) and hematological parameters, was highlighted with results presented for 6 days post-surgery during the rat&#39;s recovery. The drug-plasma concentration-time profile of orally administered natural phenol ellagic acid was determined in the JVC rat model.

Detailed microsurgical techniques are demonstrated to establish a longer-term jugular vein cannulation rat model for sequential blood collection in the same animal. Physiological and hematological parameters have been monitored during the rat's recovery phase. This model has been applied to study pharmacokinetics of orally administered polyphenol without inducing animal stress.

建立大鼠永久性颈静脉插管的显微外科技能，用于口服药物的连续血液采样

Blood sampling in small laboratory animals is necessary for pharmaceutical lead optimization but can cause great harm and stress to experimental animals, ...

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Subtitles

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样