我们感谢兽医, 护士和动物饲养者在生物医学研究中心的持续支持, 在我们的研究项目。作者承认本议定书所有审评员的重要投入。其中一些研究由奥地利胃肠病和肝病学会的 "青年科学奖" (ÖGGH) 到 PS 和奥地利内科医学协会的 "斯柯达奖" 资助。

这里描述的所有方法都已获得维也纳医科大学道德委员会和奥地利科学、研究和经济部 (BMWFW) 的批准。由于血流动力学测量代表手术干预, 必须在手术室或类似清洁工作区的无菌条件下执行程序。一般情况下, 建议在无菌条件下工作。当使用吸入麻醉时, 考虑适当的通气手术室的工作安全。在本议定书中提出的所有血流动力学读数的情况下, 必须考虑 40–50 min/动物的时间段。
1. 手术前的准备工作
<ol><li>根据制造商的指示, 打开并校准包括压力传感器在内的电子多通道录音机。</li>
	<li>将超声波流量探头 (1 毫米和2毫米) 连接到桥放大器。</li>
	<li>准备一个无菌的生理盐水的水库, 加热到体温, 37 °c, 为滋润组织或纱布压缩。</li>
	<li>记录动物体重, 提供体重调节麻醉和血流动力学参数的规范化, 按体重。</li>
	<li>准备所有的吸入麻醉设备。 注: 如果吸入麻醉和/或必要的设备不在现场或可行, 然后使用氯胺酮和甲苯噻嗪注射麻醉可以使用 (80–100毫克/千克氯胺酮与5–10毫克/千克甲苯噻嗪在生理盐水溶液, 腹腔 (ip))。对氯胺酮 (减少剂量的20–30毫克/千克, 肌内 (即时)) 后第30-45 分钟是必要的, 以诱导连续手术-平面麻醉。</li>
	<li>使用吸入麻醉感应箱 (5 分钟, 5% 伏异氟醚, 3–4 L O2流) 将动物置于短期异氟醚麻醉下。小心地倾斜箱子并且检查麻醉的深度由动物的静止状态。</li>
	<li>
用合适的自制气管插管大鼠。 注: 对于气管插管技术新的人员, 在短期吸入麻醉后, 可以给予镇痛/麻醉 (步骤 1.10), 以允许额外的插管时间。另一种选择是用于吸入麻醉的动物专用面罩。<ol>
<li>使用经改良的外周静脉导管 (14 克) 自行制作的气管插管。切断处理翼, 贴上固定在大鼠脸颊上的胶带环, 防止气管内管脱位 (图 1A)。</li>
		<li>使用自制的导丝装置, 将经改进的动脉套管作为导线架和合适的钝尖导线 (图 1B)。</li>
		<li>使用合适的插管台进行适当的动物定位。将动物置于仰卧位, 头部处于倾斜位置。 注: 如果没有插管台可用, 可以放置在仰卧位置的动物, 颈部仔细伸展在桌子的边缘。但是, 由于受伤的风险增加, 建议不要使用此程序。</li>
		<li>在老鼠一侧的门牙上固定一条缝合线, 通过将缝合线绑在另一侧 (图 1C) 轻轻地伸展动物的颈部。</li>
		<li>用聚焦光束照亮气管前侧领区。特别是在白化动物, 确保声带通过皮肤照明, 以改善可视化和快速插管。 注: 使用动物专用喉镜插管色素动物。</li>
		<li>抓住舌头, 用两个手指轻轻地拉它。</li>
		<li>使用利多卡因 (泵喷雾) 处理的棉花芽仔细麻醉喉区。</li>
		<li>使用导丝装置 (图 1D) 的支持, 在声带和气管之间插入气管导管, 将该动物插管。</li>
		<li>卸下导丝装置。</li>
		<li>把管子连接到通风机上。</li>
		<li>使用适当的动物设置启动呼吸机 (1 升/分2流, 自动流动 = 90/分钟; 吸气压力:18 mmHg;窥视: 3 mmHg, I/E = 1:2) 和检查正确插管。 注: 如果注意到胃的膨胀, 请取出管子, 再试一次。另外, 将呼吸活动与呼吸机节律进行比较, 或将两个手指放在腹部壁上, 以评估胃的潜在通胀。</li>
	</ol>
</li>
	<li>在成功插管后立即启动异氟醚麻醉, 0.5–1.0% v/v 和1升/分2-流 (图 2A)。</li>
	<li>通过颊部的 transbuccal 缝合固定气管导管, 贴附胶带环。</li>
	<li>用两个1毫升注射器 (如氯胺酮 (100 毫克/千克) (23 克套管) [或通过在骶大腿肌肉 (30 克套管) 的双侧注射中分配注射容积 (剂量)] 和 piritramide (2mg/千克) 经皮下 (南卡罗来纳州) 注射 (23 克套管)。请注意每个注射部位的最佳即时注射量 (图 2B)。</li>
	<li>涂抹眼膏。把身体的毛发夹在腹部和大腿内侧。消毒皮肤。 注: 两大腿内侧应剃须, 以允许使用对侧股动脉的 HR 和地图测量的情况下, 股动脉插管失败的一方。在以后的时间点刮胡子可能会导致头发污染手术领域。</li>
	<li>用胶带固定在加热垫 (38 °c) 的仰卧位上的动物 (图 2C)。</li>
	<li>通过使用直肠温度探针 (图 2D) 连续监测动物的体温。</li>
	<li>在任何干预或手术前, 通过盖子闭合反射和脚趾捏试验评估麻醉深度。</li>
</ol>2. 人力资源和地图的测量
<ol><li>切开皮肤的大腿内侧 (选择一侧) 以上的假设位置的股动脉通过解除皮的组织钳和删除皮肤面积约2厘米的长度由梅奥剪刀 (图 3A)。</li>
	<li>公开和坦率地解剖包括股动脉的结缔组织 (膜) 的动脉-静脉-神经复合体, 反复打开止血沿复杂。</li>
	<li>从周围组织沿 ~ 1–1.5 厘米解剖动脉-静脉-神经复合体 (图 3B)。</li>
	<li>如有必要, 删除皮下/周围脂肪, 以更好地查看和解剖。 注意: 用角质层剪刀去除皮下/周围脂肪时要小心, 因为受伤的血管可能会引起 bleedings。如果发现出血, 请使用小纱布在出血区施加压力, 或用止血止血。</li>
	<li>用两个高精度45°角宽点钳将股动脉与股静脉和神经分开 (认为股静脉是最内侧的结构, 股动脉位于更侧面) (图 3C-F)。</li>
	<li>在股动脉尽可能远端放置结扎, 并在缝线上使用弯曲钳对股动脉施加温和的纵向牵引。</li>
	<li>在股动脉上放置第二个预打结 (但不是闭合结) 缝合 (图 3G)。</li>
	<li>准备一个合适的导管 (PE-50 大鼠股动脉) 与倾斜 (~ 45°) 切断尖端冲洗与血气不育的生理盐水溶液 (5 毫升注射器和23克钝套管)。确保导管内没有气泡, 因为它们阻碍了动脉压力和 HR 读数 (图 3H)。</li>
	<li>用微钳压缩外露部分近端的股动脉, 暂时停止动脉血流 (图 3I)。</li>
	<li>使用弯曲套管 (23 克), 同时在股动脉下方放置支撑微金属刮刀 (图 3J), 在解剖切片的远端处穿孔动脉壁。</li>
	<li>仔细 catheterize 股动脉通过穿孔与倾斜尖端的导管朝向向上。提前导管直到微钳接近 (图 3K)。 注: 如果第一次尝试不成功的股动脉插管, 可以进行更近端的第二次尝试 (从步骤2.8 开始)。如果动脉破裂或严重出血发生, 结扎或夹紧动脉尽可能近, 以防止进一步失血。如果失血量最小, 尝试置管对侧股动脉 (从步骤2.1 开始)。</li>
	<li>打开微钳, 检查脉搏动脉血流进入导管 (图 3L)。</li>
	<li>通过阻断导管流出, 防止血液进一步流入导管。</li>
	<li>将导管固定在其腔内位置, 方法是关闭股骨动脉周围的准备好的结扎管 (图 3M)。</li>
	<li>反复冲洗和吸入股动脉导管, 以评估适当的血管内放置。吸入血柱中的动脉搏动容易引起注意。</li>
	<li>用远端结扎末端固定导管位置, 以防止脱位并确保导管的纵向位置 (图 3N)。</li>
	<li>另外, 将导管紧贴在手术台上的动物身上, 以确保其安全并防止意外脱位。</li>
	<li>将充满生理盐水溶液的导管连接到压力传感器, 同时避免气泡的形成。</li>
	<li>使用数字接口开始记录 HR 和地图 (图 3O)。</li>
	<li>用小的湿纱布覆盖内侧大腿外露的区域 (图 3P)。</li>
	<li>计算存在高动力 (hd) 索引: hd = 地图/HR。 注: 在高级 PHT 中, HD 指数与非门脉高压动物相比升高。然而, 手术中 HD 指数的增加也可能表明出血、低血容量或疼痛。如果记录的地图值很低, 但信号良好, 脉搏流在导管中检测到, 检查麻醉的水平, 并有可能降低麻醉水平。不要完全停止异氟醚的麻醉, 因为这可能导致麻醉深度不足, 根据动物福利和良好的科学实践。</li>
</ol>3. 肠系膜上动脉血流 (SMABF)
<ol><li>
执行中位剖腹手术 (图 4A-C)
	<ol>
<li>抬起皮肤层与组织钳5–6厘米以下的剑突, 并删除一个薄条皮肤使用梅奥剪刀上面的 linea, 直到剑突到达。</li>
		<li>在中间的皮肤切口, 抬起肌肉层的组织钳沿 linea, 创造距离之间的腹壁和内脏器官。</li>
		<li>用手术刀在 linea 上切开腹腔, 打开腹腔腔。扩大开放, 同时解除腹壁的组织钳与 Metzenbaum 剪刀沿 linea 与皮肤层相同的距离。</li>
	</ol>
</li>
	<li>小心地挖掘肠道使用湿棉花芽开始与 coecum, 并把它放在一个大纱布压缩浸泡在无菌的生理盐水溶液旁边的切口 (图 4D-F)。</li>
	<li>将肠道包裹在纱布中, 确保它与无菌的生理盐水溶液湿润 (图 4G)。</li>
	<li>定位并暴露肠系膜上动脉。</li>
	<li>解剖肠系膜上动脉两个自制钝 ' Schwabl ' 钩: 抬起动脉的第一个钩, 并试图把第二个通过同一组织隧道。将肠系膜上动脉暴露在5毫米的距离, 以确保流探针 (1 毫米) 可以放置在它周围 (图 4H-K)。 注: 如果首选, 一个精确的45°角宽点钳可以用来解除动脉。' Schwabl '-钩是准备从30克套管与坦率折断的小贴士弯曲到钩形。如果侧支动脉是广泛的, "Schwabl" 钩可能是安全的 bleedings 从络, 而解剖肠系膜上动脉可以避免。如果在准备肠系膜上动脉时出血, 请在出血部位放置一个小纱布, 以温和的压力。小出血通常会很快停止;时刻牢记要定期滋润组织 (步骤 1.12)。如果肠系膜上动脉自身受损, 血流动力学评估必须终止。</li>
	<li>将超声凝胶应用于超声流量探头的传感器, 并将其附着于内脏肠系膜动脉。将其与肠系膜上动脉的自然路线 (图 4L M) 对齐。</li>
	<li>关闭流量探头 (1 毫米), 如果需要, 在多普勒传感器上轻轻地应用额外的超声波凝胶, 以提高信号质量。使用注射器 (20 毫升) 填充超声凝胶和钝尖套管 (18 克) (图 4N-O)。 注: 如果水流探针在容器的自然过程中没有很好地排列, 张力可能导致血管收缩因而, 湍流流动, 减少流量测量的准确性。试着将水流探头的方向沿着容器的自然路径重新对准, 然后在适当的位置上充分固定流量探头。</li>
	<li>测量 SMABF 并评估脉搏流信号对股动脉记录的收缩峰值的符合性。 注: 如果记录的地图值非常低, 但信号良好, 脉搏流在导管中检测到, 检查麻醉水平, 并有可能降低麻醉水平。不要完全停止异氟醚的麻醉, 因为这可能导致麻醉深度不足, 根据动物福利和良好的科学实践。</li>
	<li>找到流量探头 (1 毫米) 的稳定位置, 并固定流探头的电缆。开始录制 SMABF, 而不进一步操作流探针 (1 毫米) (图 4P)。</li>
</ol>4. PVBF
<ol><li>在靠近肝门的肠系膜背面定位并暴露门静脉 (图 5A)。</li>
	<li>使用高精度45°角宽点钳从周围组织中轻轻地解剖门静脉: 将门静脉轻轻地和反复地推到门静脉下以创建组织隧道 (图 5B)。 注: 如果门静脉周围组织在准备门静脉时发生出血, 应使用棉花芽对出血部位施加温和压力1-2 分钟;这经常会止住流血。</li>
	<li>通过打开高精度45°角宽点钳慢慢扩大隧道, 并将门静脉沿5-6 毫米的距离暴露, 允许放置周围流探头 (2 毫米) (图 5C, D)。</li>
	<li>将超声波凝胶应用于超声波流量探头的传感器上, 并将其连接到与其自然路径对齐的门静脉 (图 5E)。</li>
	<li>如前所述 (步骤 3.7), 关闭流探头 (2 毫米) 并应用额外的超声波凝胶 (图 5F)。</li>
	<li>确保流探头以非紧缩的方式放置在门静脉周围 (图 5G)。 注: 如果水流探针在容器的自然过程中没有很好地排列, 张力可能导致血管收缩因而, 湍流流动, 减少流量测量的准确性。试着将水流探头的方向沿着容器的自然路径重新对准, 然后在适当的位置上充分固定流量探头。</li>
	<li>找到流量探头 (2 毫米) 的稳定位置, 并固定流探头的电缆。然后, 开始录制 PVBF (图 5H)。 注: 如果记录的地图值非常低, 但信号良好, 脉搏流在导管中检测到, 检查麻醉水平, 并有可能降低麻醉水平。不要完全停止异氟醚的麻醉, 因为这可能导致麻醉深度不足, 根据动物福利和良好的科学实践。</li>
</ol>5. PP
<ol><li>准备一个导管 (PE-50 大鼠肠系膜静脉) 与倾斜 (约 45°) 切断尖端, 是冲洗与无菌的生理盐水溶液 (5 毫升注射器和钝23克套管)。注意, 导管内没有气泡, 因为它们阻碍了 PP 读数 (图 6A)。</li>
	<li>用湿手套处理肠道, 把它分散在手指上 (图 6B)。</li>
	<li>最优化的观点, 肠系膜血管床接近小肠 (图 6C)。</li>
	<li>确定主要静脉肠系膜血管导致门静脉 (腔 ileocolica-腔肠系膜上腔 portae)。</li>
	<li>找到合适的 ileocolonic 静脉连接处, 可供插管使用。</li>
	<li>首先将导管插入肠系膜组织中, 将肠系膜上的内脏腹膜穿透到选择导尿管的血管连接处。</li>
	<li>小心地推进导管的倾斜尖端靠近回结肠静脉的交界处, 直到看到船只交界处的轻微印象 (图 6D)。</li>
	<li>最后, catheterize 血管壁在血管的交叉角上穿孔, 使静脉系统与连接血管路径一致。(图 6E)。 注: 如果出血发生在回结肠静脉插管, 使用拇指压机与小纱布压缩在出血区。应保持这种压力, 使其能止血。然后, 尝试将导管插入回结肠静脉的更近端分支。</li>
	<li>将导管小心地向前推进, 沿着主静脉血管通路到门静脉, 不穿孔静脉 (图 6F)。 注: 将导管保持在足够的距离, 使其位于门静脉主分支周围的流探针上, 以避免流信号中的工件, 防止门静脉穿孔。</li>
	<li>将充满生理盐水溶液的导管连接到压力传感器, 同时避免气泡的形成。</li>
	<li>开始录制 PP。</li>
	<li>在稳定条件下同时记录所有血流动力学参数 (图 6G, H)。可选的是, 门静脉导管可以固定到位使用组织胶水和肠道可以重新定位到腹腔。 注: 如果记录的地图值非常低, 但信号良好, 脉搏流在导管中检测到, 检查麻醉水平, 并有可能降低麻醉水平。不要完全停止异氟醚的麻醉, 因为这可能导致麻醉深度不足, 根据动物福利和良好的科学实践。</li>
</ol>6. IHVR
<ol><li>在牺牲了动物之后, 测量肝脏的重量。计算 IHVR: IHVR = PVBF/PP. 将这个 PVBF 值正常化为肝脏的重量。</li>
</ol>

<ol>
	<li>Ripoll, C., 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=Hepatic+venous+pressure+gradient+predicts+clinical+decompensation+in+patients+with+compensated+cirrhosis.">Hepatic venous pressure gradient predicts clinical decompensation in patients with compensated cirrhosis.</a> Gastroenterology. 133 (2), 481-488 (2007).</li><li>Bosch, J., Groszmann, R. J., Shah, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+in+the+understanding+of+the+pathophysiological+basis+of+portal+hypertension:+How+changes+in+paradigm+are+leading+to+successful+new+treatments.">Evolution in the understanding of the pathophysiological basis of portal hypertension: How changes in paradigm are leading to successful new treatments.</a> J Hepatol. 62, S121-S130 (2015).</li><li>Blachier, M., Leleu, H., Peck-Radosavljevic, M., Valla, D. C., Roudot-Thoraval, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+burden+of+liver+disease+in+Europe:+a+review+of+available+epidemiological+data.">The burden of liver disease in Europe: a review of available epidemiological data.</a> J Hepatol. 58 (3), 593-608 (2013).</li><li>Colle, I., Geerts, A. M., Van Steenkiste, C., Van Vlierberghe, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamic+Changes+in+Splanchnic+Blood+Vessels+in+Portal+Hypertension.">Hemodynamic Changes in Splanchnic Blood Vessels in Portal Hypertension.</a> Advances in Integrative Anatomy and Evolutionary Biology. 291 (6), 699-713 (2008).</li><li>Laleman, W., Van Landeghem, L., Wilmer, A., Fevery, J., Nevens, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Portal+hypertension:+from+pathophysiology+to+clinical+practice.">Portal hypertension: from pathophysiology to clinical practice.</a> Liver International. 25 (6), 1079-1090 (2005).</li><li>Franchis, R. d. . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Updating+Consensus+in+Portal+Hypertension:+Report+of+the+Baveno+III+Consensus+Workshop+on+definitions,+methodology+and+therapeutic+strategies+in+portal+hypertension.">Updating Consensus in Portal Hypertension: Report of the Baveno III Consensus Workshop on definitions, methodology and therapeutic strategies in portal hypertension.</a> Journal of Hepatology. 33 (5), 846-852 (2000).</li><li>Zardi, E. M., Di Matteo, F. M., Pacella, C. M., Sanyal, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invasive+and+non-invasive+techniques+for+detecting+portal+hypertension+and+predicting+variceral+bleeding+in+cirrhosis:+a+review.">Invasive and non-invasive techniques for detecting portal hypertension and predicting variceral bleeding in cirrhosis: a review.</a> Annals of medicine. 46 (1), 8-17 (2014).</li><li>Kumar, A., Sharma, P., Sarin, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+venous+pressure+gradient+measurement:+time+to+learn.">Hepatic venous pressure gradient measurement: time to learn.</a> Indian J Gastroenterol. 27 (2), 74-80 (2008).</li><li>Tsochatzis, E. A., Bosch, J., Burroughs, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+cirrhosis.">Liver cirrhosis.</a> Lancet. 383 (9930), 1749-1761 (2014).</li><li>Abraldes, J. G., Pasar&#237;n, M., Garc&#237;a-Pag&#225;n, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+portal+hypertension.">Animal models of portal hypertension.</a> World Journal of Gastroenterology : WJG. 12 (41), 6577-6584 (2006).</li><li>Reiberger, T., 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=Sorafenib+attenuates+the+portal+hypertensive+syndrome+in+partial+portal+vein+ligated+rats.">Sorafenib attenuates the portal hypertensive syndrome in partial portal vein ligated rats.</a> Journal of Hepatology. 51 (5), 865-873 (2009).</li><li>Schwabl, 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=Pioglitazone+decreases+portosystemic+shunting+by+modulating+inflammation+and+angiogenesis+in+cirrhotic+and+non-cirrhotic+portal+hypertensive+rats.">Pioglitazone decreases portosystemic shunting by modulating inflammation and angiogenesis in cirrhotic and non-cirrhotic portal hypertensive rats.</a> Journal of Hepatology. 60 (6), 1135-1142 (2014).</li><li>Reiberger, T., 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=Nebivolol+treatment+increases+splanchnic+blood+flow+and+portal+pressure+in+cirrhotic+rats+via+modulation+of+nitric+oxide+signalling.">Nebivolol treatment increases splanchnic blood flow and portal pressure in cirrhotic rats via modulation of nitric oxide signalling.</a> Liver International. 33 (4), 561-568 (2013).</li><li>Schwabl, 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+FXR+agonist+PX20606+ameliorates+portal+hypertension+by+targeting+vascular+remodelling+and+sinusoidal+dysfunction.">The FXR agonist PX20606 ameliorates portal hypertension by targeting vascular remodelling and sinusoidal dysfunction.</a> Journal of Hepatology. 66 (4), 724-733 (2017).</li><li>Mandorfer, 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=Sustained+virologic+response+to+interferon-free+therapies+ameliorates+HCV-induced+portal+hypertension.">Sustained virologic response to interferon-free therapies ameliorates HCV-induced portal hypertension.</a> J Hepatol. 65 (4), 692-699 (2016).</li><li>Schwabl, 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=Interferon-free+regimens+improve+portal+hypertension+and+histological+necroinflammation+in+HIV/HCV+patients+with+advanced+liver+disease.">Interferon-free regimens improve portal hypertension and histological necroinflammation in HIV/HCV patients with advanced liver disease.</a> Aliment Pharmacol Ther. 45 (1), 139-149 (2017).</li><li>Reiberger, T., Mandorfer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta+adrenergic+blockade+and+decompensated+cirrhosis.">Beta adrenergic blockade and decompensated cirrhosis.</a> Journal of Hepatology. 66 (4), 849-859 (2017).</li><li>Reiberger, T., 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=Carvedilol+for+primary+prophylaxis+of+variceal+bleeding+in+cirrhotic+patients+with+haemodynamic+non-response+to+propranolol.">Carvedilol for primary prophylaxis of variceal bleeding in cirrhotic patients with haemodynamic non-response to propranolol.</a> Gut. 62 (11), 1634-1641 (2013).</li><li>Reiberger, T., 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=Austrian+consensus+guidelines+on+the+management+and+treatment+of+portal+hypertension+(Billroth+III).">Austrian consensus guidelines on the management and treatment of portal hypertension (Billroth III).</a> Wiener klinische Wochenschrift. 129 (3), 135-158 (2017).</li><li>de Franchis, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expanding+consensus+in+portal+hypertension.">Expanding consensus in portal hypertension.</a> Journal of Hepatology. 63 (3), 743-752 (2015).</li><li>Pinter, 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=The+effects+of+sorafenib+on+the+portal+hypertensive+syndrome+in+patients+with+liver+cirrhosis+and+hepatocellular+carcinoma+-+a+pilot+study.">The effects of sorafenib on the portal hypertensive syndrome in patients with liver cirrhosis and hepatocellular carcinoma - a pilot study.</a> Alimentary Pharmacology &amp; Therapeutics. 35 (1), 83-91 (2012).</li><li>Schwabl, P., Laleman, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+treatment+options+for+portal+hypertension.">Novel treatment options for portal hypertension.</a> Gastroenterol Rep (Oxf). 5 (2), 90-103 (2017).</li><li>Klein, S., Schierwagen, R., Uschner, F., Trebicka, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mouse and Rat Models of Induction of Hepatic Fibrosis and Assessment of Portal Hypertension. , (2017).</li><li>Russell, W. M. S., Burch, 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=.">.</a> The Principles of Humane Experimental Technique. , (1959).</li><li>Langhans, W., Myrtha, A., Riediger, T., Lutz, T. 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> Routine animal use procedures. , (2016).</li><li>Animal Care and Use Program. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rat and Mouse anesthesia and analgesia: Formulary and General Drug Information. , (2016).</li><li>Davis, J. 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> Current Protocols in Neuroscience. , (2001).</li><li>Albrecht, M., Henke, J., Tacke, S., Markert, M., Guth, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+isoflurane,+ketamine-xylazine+and+a+combination+of+medetomidine,+midazolam+and+fentanyl+on+physiological+variables+continuously+measured+by+telemetry+in+Wistar+rats.">Effects of isoflurane, ketamine-xylazine and a combination of medetomidine, midazolam and fentanyl on physiological variables continuously measured by telemetry in Wistar rats.</a> BMC Veterinary Research. 10 (1), 198 (2014).</li><li>Redfors, B., Shao, Y., Omerovic, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+anesthetic+agent,+depth+of+anesthesia+and+body+temperature+on+cardiovascular+functional+parameters+in+the+rat.">Influence of anesthetic agent, depth of anesthesia and body temperature on cardiovascular functional parameters in the rat.</a> Laboratory Animals. 48 (1), 6-14 (2014).</li><li>Becker, 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=.">.</a> Statement on anesthesia methodologies: Recommondations on anaesthesia methodologies for animal experimentation in rodents and rabbits. , (2016).</li></ol>

作者没有什么可透露的。

PP 是评价门脉高压综合征的主要结局参数, 反映肝硬化的严重程度。基质沉积 (即纤维化) 和正弦血管收缩 (由于血管收缩剂肝表达的增加和对血管扩张剂的反应减少) 导致 IHVR 增加。聚丙烯的重要性和它对慢性肝病的影响已显示在多个临床前11,12,13,14和临床研究15,<sup cla...

PHT 被定义为门静脉系统的病理增加的血压, 可能导致严重并发症的肝硬化患者, 如静脉曲张出血和腹水1。肝前 (如门静脉血栓) 和肝后 (如布-加综合征) PHT 少见, 肝硬化肝内 PHT 是 PHT2的最常见病因。
在肝硬化中, PP 主要是由于升高的 IHVR3而增加的。在晚期, PHT 的增加 PVBF 由于增加的心脏输出和减少系统性和内脏血管阻力-定义门脉高压综合征4。欧姆定律 (ΔP = Q R) 意味着 IHVR 和血流与 PP5成正比。在患者中, 直接测量 PP 是危险的, 没有例行执行;相反, 肝静脉压力梯度 (HVPG) 被用作间接测量 PP6,7。HVPG 的计算方法是从楔形肝静脉压 (WHVP) 中减去游离肝静脉压 (FHVP), 用放置在肝静脉中的气囊导管测量8。1–5 mmHg 之间的生理 HVPG 范围, 而 HVPG ≥10 mmHg 定义临床重要的门静脉高压 (CSPH), 表明 PHT 相关并发症的风险增加, 如静脉曲张出血, 腹水和肝性脑病9.尽管 PP (即HVPG) 是 PHT 严重性的最相关参数, 但有关 PHT 其他成分的信息, 包括存在高动力循环的严重性 (HR、MAP)、内脏/肠系膜动脉血流和 IHVR, 对于全面了解 PHT 的基本机制。
因此, 与间接测量人的 pp 相比, 引入的方法为大鼠提供了直接测量 pp 的优势, 并允许记录额外的血流动力学参数表征门脉高压综合征。此外, 直接测量 PP 是一个很好的综合读数的肝纤维化的数量 (IHVR 的一个主要决定因素), 克服了一定程度的纤维化量化相关的肝组织抽样错误。
肝硬化 PHT 最常用的啮齿动物模型包括手术胆管结扎术 (BDL)、毒素诱发肝损伤 (即四氯化碳、硫乙酰胺或硝胺管理) 和饮食诱导的代谢性肝。疾病模型。肝前性 (非肝硬化) PHT 可由部分门静脉结扎术 (PPVL)10引起。
小啮齿类动物非常适合所提出的方法, 包括老鼠, 仓鼠, 大鼠, 或兔子, 并与相对较低的维护成本。尽管所有的血流动力学评估是可行的, 在小鼠, 更好的准确性和重现性的结果被发现与大鼠或较大的啮齿动物由于明显的优势, 动物的大小。此外, 还需要特定的微仪器和器件来获得类似的小鼠血流动力学参数。最后, 大鼠更健壮, 相关的发病率和死亡率较低, 因此, 大鼠的辍学率可能低于小鼠。
所提出的方法非常适合评估肝病的具体治疗 (即抗纤维化或消炎药) 或新的药理学方法, 影响血管张力和/或内皮生物学;因此, 可能影响 PHT 的血流动力学参数。

<table><tbody><tr><td>&lt;strong&gt;Instruments&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>LabChart 7 Pro software</td><td>ADInstruments, Colorado Springs, CO, USA</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Software&lt;/em&gt;</td></tr><tr><td>ML870 PowerLab 8/30</td><td>ADInstruments, Colorado Springs, CO, USA</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Electronic multichannel recorder&lt;/em&gt;</td></tr><tr><td>MLT0380/D</td><td>ADInstruments, Colorado Springs, CO, USA</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Pressure transducer (x2: for Portal Pressure and Arterial Pressure)&lt;/em&gt;</td></tr><tr><td>ML112 Quad Bridge Amplifier</td><td>ADInstruments, Colorado Springs, CO, USA</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Bridge amplifier&lt;/em&gt;</td></tr><tr><td>TS420</td><td>Transonic Systems Inc., Ithaca, NY, USA</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Flowmeter module&lt;/em&gt;</td></tr><tr><td>Biological Research Apparatus 7025</td><td>UGO BASILE S.R.L., Comerio, Italy</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Ventilator&lt;/em&gt;</td></tr><tr><td>Vapor 2000</td><td>Dr&amp;auml;ger Medical AG &amp;amp; Co. KG, L&amp;uuml;beck, Germany</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Isofluran Vaporizer&lt;/em&gt;</td></tr><tr><td>Perivascular probes (rat) for Transonic systems (Superior Mesenteric Artery)</td><td>Transonic Systems Inc., Ithaca, NY, USA</td><td>#MA1PRB</td><td>&lt;em&gt;Ultrasonic flow probe (1mm)&lt;/em&gt;</td></tr><tr><td>Perivascular probes (rat) for Transonic systems (Portal Vein)</td><td>Transonic Systems Inc., Ithaca, NY, USA</td><td>#MA2PSB</td><td>&lt;em&gt;Ultrasonic flow probe (2mm)&lt;/em&gt;</td></tr><tr><td>1st for intubation &amp;amp; 2nd for clean skin incisions</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Mayo scissor [x2]&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;Metzenbaum scissor&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Cuticle scissor&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>e.g. Adson Brown tissue forceps</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Tissue Forceps&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;High precision 45&amp;deg; angle broad point forceps [x2]&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Hemostat [x4]&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>e.g. Mikulicz peritoneal clamp</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Curved clamp&lt;/em&gt;</td></tr><tr><td>e.g. Dieffenbach clamp</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Micro clamp&lt;/em&gt;</td></tr><tr><td>e.g. micro spatula with flat ends, width 4 mm,</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Micro metal spatula&lt;/em&gt;</td></tr><tr><td>for transbuccal suture at intubation</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Needle holder&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;Scalpel grip&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>selfmade</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Intubation desk&lt;/em&gt;</td></tr><tr><td>blut, flexible and with a suitable diameter for arterial cannula and venflow</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Blunt steel wire&lt;/em&gt;</td></tr><tr><td>modified arterial line 20G with Flowstich</td><td>Becton Dickinson, Farady Road, Swindon, UK</td><td>#682245</td><td>&lt;em&gt;Arterial line&lt;/em&gt;</td></tr><tr><td>&lt;em&gt;Heating pad&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Rectal temerature probe&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Saline heater&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Laryngoscope (specific for animal size, e.g. rat)&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Inductionbox for inhalation anesthesia&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Scale (able to measure mg)&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;em&gt;Hair clipper&lt;/em&gt;</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td><td>&amp;nbsp;-</td></tr><tr><td>&lt;strong&gt;Name &lt;/strong&gt;</td><td>&lt;strong&gt;Company &lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Consumables &lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>e.g. modified BD Venflon Pro Safety 14GA</td><td>Becton Dickinson Infusion Therapy, AB, SE251 06 Helsingborg, Sweden</td><td>#393230</td><td>&lt;em&gt;Peripheral venous catheter (14G)&lt;/em&gt;</td></tr><tr><td>Fine-Bore Polyethylene Tubing, ID 0.58mm, OD 0.96mm, Portex,</td><td>Smiths Medical International Ltd., Kent, UK</td><td>#800/100/200</td><td>&lt;em&gt;Catheter tube (PE-50)&lt;/em&gt;</td></tr><tr><td>e.g. Omnifix-F Solo</td><td>B. Braun Melsungen AG, Melsungen, Germany</td><td>#9161406V</td><td>&lt;em&gt;Syringe 1mL&lt;/em&gt;</td></tr><tr><td>e.g. Injekt Solo</td><td>B. Braun Melsungen AG, Melsungen, Germany</td><td>#4606051V</td><td>&lt;em&gt;Syringe 5mL&lt;/em&gt;</td></tr><tr><td>e.g. Injekt Solo</td><td>B. Braun Melsungen AG, Melsungen, Germany</td><td>#4606205V</td><td>&lt;em&gt;Syringe 20mL&lt;/em&gt;</td></tr><tr><td>e.g. BD Microlance 3, 18G - 1 1/2&quot;</td><td>Becton Dickinson S.A., Fraga, Spain</td><td>#304622</td><td>&lt;em&gt;Cannula (18G)&lt;/em&gt;</td></tr><tr><td>e.g. BD Microlance 3, 23G - 1&quot;</td><td>Becton Dickinson S.A., Fraga, Spain</td><td>#300800</td><td>&lt;em&gt;Cannula (23G)&lt;/em&gt;</td></tr><tr><td>e.g. BD Microlance 3, 30G - 1/2&quot;</td><td>Becton Dickinson S.A., Fraga, Spain</td><td>#304000</td><td>&lt;em&gt;Cannula (30G) &lt;/em&gt;</td></tr><tr><td>e.g. Leukoplast S</td><td>BSN medical GmbH, Hamburg,&amp;nbsp; Germany</td><td>#47619-00</td><td>&lt;em&gt;Adhesive tape&lt;/em&gt;</td></tr><tr><td>e.g. Gazin RK Mullkompressen (18x8cm)</td><td>Lohmann &amp;amp; Rauscher, Vienna, Austria</td><td>#10972</td><td>&lt;em&gt;Gauze compress (small)&lt;/em&gt;</td></tr><tr><td>e.g. Gazin RK Mullkompressen (5x5cm)</td><td>Lohmann &amp;amp; Rauscher, Vienna, Austria</td><td>#10961</td><td>&lt;em&gt;Gauze compress (big)&lt;/em&gt;</td></tr><tr><td>Silk Braided black, USP 4/0, EP 1.5</td><td>SMI AG, St. Vith, Belgium</td><td>#2021-04</td><td>&lt;em&gt;Suture (Silk 4/0, EP 1.5)&lt;/em&gt;</td></tr><tr><td>e.g. Mersilk, 2-0 (3 Ph. Eur.), PS-1 Prime</td><td>Johnson &amp;amp; Johnson Medical GmbH - Ethicon Deutschland, Germany</td><td>#EH7552</td><td>&lt;em&gt;Transbuccal suture&lt;/em&gt;</td></tr><tr><td>e.g. Cottonbuds (2.2mm, 15cm)</td><td>Paul Hartmann AG, Heidenheim, Germany</td><td>#967936</td><td>&lt;em&gt;Cotton buds&lt;/em&gt;</td></tr><tr><td>e.g. Vue Ultrasoundgel</td><td>Optimum Medical Limited, UK</td><td>#1157</td><td>&lt;em&gt;Ultrasound gel&lt;/em&gt;</td></tr><tr><td>e.g. Glubran 2</td><td>Gem srl, Viareggio, Italy</td><td>#G-NB2-50</td><td>&lt;em&gt;Tissue glue&lt;/em&gt;</td></tr><tr><td>e.g. Surgical scalpell knife Nr. 10 - carbon steel</td><td>Swann-Morton, England, B.S.</td><td>#202</td><td>&lt;em&gt;Scalpel Knife&lt;/em&gt;</td></tr><tr><td>Heparin, 5000 i.E./mL (Natriumheparin)</td><td>Medicamentum Pharma GmbH, Allerheiligen im M&amp;uuml;rztal, Austria</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Heparin&lt;/em&gt;</td></tr><tr><td>Florane</td><td>Aesica Queenborough Ltd., Queenborough, UK</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Isoflurane&lt;/em&gt;</td></tr><tr><td>OeloVital (5g)</td><td>Fresenius Kabi Austira Gmbh, Graz, Austria</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Eye gel&lt;/em&gt;</td></tr><tr><td>Ketasol</td><td>aniMedica GmbH, Senden-B&amp;ouml;sensell, Germany</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Ketamine&lt;/em&gt;</td></tr><tr><td>Rompun</td><td>Bayer Austria Ges.m.b.H., Vienna, Austria</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Xylazine&lt;/em&gt;</td></tr><tr><td>Xylocain 10% Pumpspray</td><td>AstraZeneca &amp;Ouml;sterreich GmbH, Vienna, Austria</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Lidocaine pump spray&lt;/em&gt;</td></tr><tr><td>Dipidolor</td><td>Jansen-Cilag Pharma GmbH, Vienna, Austria</td><td>&amp;nbsp;-</td><td>&lt;em&gt;Piritramide&lt;/em&gt;</td></tr><tr><td>NaCl 0.9% Fresenius, 1L</td><td>Fresenius Kabi Austira GmbH, Graz, Austria</td><td>#13LIP132</td><td>&lt;em&gt;Physiological saline solution&lt;/em&gt;</td></tr></tbody></table>

invasive hemodynamic characterization of the portal-hypertensive syndrome in cirrhotic rats

这是一个详细的协议描述侵袭性血流动力学测量肝硬化大鼠门脉高压综合征的特点。肝硬化门静脉高压症 (PHT) 是导致肝病患者最严重并发症的原因。门脉高压综合征的全貌是由于肝内血管阻力增加 (IHVR), 存在高动力循环, 内脏血流量增加, 门静脉压力增加 (PP)。进展性内脏动脉舒张和增加心脏输出与高心率 (HR), 但低动脉压力特点的门脉高压综合征。
目前正在开发新的治疗方法, 目的是降低 PP 的靶向 IHVR 或增加内脏血流-但副作用的系统性血流动力学可能会发生。因此, 需要对门静脉、内脏和全身血流动力学参数进行详细的描述, 包括测量 PP、门静脉血流量 (PVBF)、肠系膜动脉血流量、平均动脉压 (MAP) 和 HR 是临床前PHT 新疗法的疗效评价。我们的视频文章为读者提供了一个结构化的协议来执行肝硬化大鼠的侵袭性血流动力学测量。特别是, 我们描述了通过回结肠静脉插管的股动脉和门静脉, 并通过周围多普勒超声流量探头测量门静脉和内脏血流量。PHT 了不同大鼠模型的代表性结果。

根据动物模型和肝脏疾病的严重性, 门脉高压综合征的 PHT 程度和严重性不同 (图 7)。
BDL 模型导致胆汁性肝硬化胆汁淤积。因此, 随着时间的推移, PP 的增加和存在高动力的循环发展, 从人力资源的增加和地图的减少看到。在肝硬化动物中, SMABF、PVBF 和 IHVR 也增加和谐肝脏和血流动力学的改变 (<strong class="xf...

Watch this Scientific Journal Video about Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats at JoVE.com

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

在这里, 我们描述了一个详细的协议, 以侵入性测量的血流动力学参数, 包括门压力, 内脏血流量, 系统血流动力学, 以表征门脉高压综合征的大鼠。

肝硬化大鼠门脉高压综合征的侵袭性血流动力学特征

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

invasive-hemodynamic-characterization-portal-hypertensive-syndrome

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

15.0K 次观看.  Medical University of Vienna, Vienna, Austria. 在这里, 我们描述了一个详细的协议, 以侵入性测量的血流动力学参数, 包括门压力, 内脏血流量, 系统血流动力学, 以表征门脉高压综合征的大鼠。

视频: Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

科研

医学

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory&#160;rate&#160;and heart&#160;rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.
This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41&#177;2.94 cmH2O which was significantly higher (P&#60;0.05) than before resection (6.87&#177;2.39 cmH2O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.

This film demonstrates how to acquire systemic and hepatic hemodynamics in mice. The whole monitoring includes acquisition of vital parameters, systemic blood pressure, central venous pressure, common hepatic artery flow rate, and portal vein pressure as well as the portal flow rate in mice.

全身及肝脏血流动力学参数的监测小鼠

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle proliferation and obliteration of pulmonary arterioles. This in turn results in right ventricular (RV) failure, with significant morbidity and mortality. Rodent models of PAH, in the mouse and the rat, are important for understanding the pathophysiology underlying this rare disease. Notably, different models of PAH may be associated with different degrees of pulmonary hypertension, RV hypertrophy and RV failure. Therefore, a complete hemodynamic characterization of mice and rats with PAH is critical in determining the effects of drugs or genetic modifications on the disease. 
Here we demonstrate standard procedures for assessment of right ventricular function and hemodynamics in both rat and mouse PAH models. Echocardiography is useful in determining RV function in rats, although obtaining standard views of the right ventricle is challenging in the awake mouse. Access for right heart catheterization is obtained by the internal jugular vein in closed-chest mice and rats. Pressures can be measured using polyethylene tubing with a fluid pressure transducer or a miniature micromanometer pressure catheter. Pressure-volume loop analysis can be performed in the open chest. After obtaining hemodynamics, the rodent is euthanized. The heart can be dissected to separate the RV free wall from the left ventricle (LV) and septum, allowing an assessment of RV hypertrophy using the Fulton index (RV/(LV+S)). Then samples can be harvested from the heart, lungs and other tissues as needed.

Pulmonary arterial hypertension (PAH) is a disease of pulmonary arterioles that leads to their obliteration and the development of right ventricular failure. Rodent models of PAH are critical in understanding the pathophysiology of PAH. Here we demonstrate hemodynamic characterization, with right heart catheterization and echocardiography, in the mouse and rat.

肺动脉高压的啮齿动物模型的血流动力学特性

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle ...

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt (MCT+Flow). The shunt is created by inserting an 18-G needle from the abdominal aorta into the adjacent caval vein. Increased pulmonary flow has been demonstrated as an essential trigger for a severe form of PAH with distinct phases of disease progression, characterized by early medial hypertrophy followed by neointimal lesions and the progressive occlusion of the small pulmonary vessels. To measure the right heart and pulmonary hemodynamics in this model, right heart catheterization is performed by inserting a rigid cannula containing a flexible ball-tip catheter via the right jugular vein into the right ventricle. The catheter is then advanced into the main and the more distal pulmonary arteries. The histopathology of the pulmonary vasculature is assessed qualitatively, by scoring the pre- and intra-acinar vessels on the degree of muscularization and the presence of a neointima, and quantitatively, by measuring the wall thickness, the wall-lumen ratios, and the occlusion score.

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

分流手术，右心导管，血管形态计量学在大鼠模型中的流量引起的肺动脉高压

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt ...

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

肝分型与门静脉结扎大鼠分期肝切除 (ALPPS) 手术模型的建立

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Measurement of the Hepatic Venous Pressure Gradient and Transjugular Liver Biopsy

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced chronic liver disease followed by an instruction for transjugular biopsy. Under local anesthesia and ultrasound guidance, a catheter introducer sheath is placed in the right internal jugular vein. Using&#160;fluoroscopic guidance, a balloon catheter is advanced into the inferior vena cava (IVC) and inserted into a large hepatic vein. Correct and sufficient wedge position of the catheter is ensured by injecting contrast media while the balloon is blocking the outflow of the cannulated hepatic vein. After calibrating the external pressure transducer, continuous pressure recordings are obtained with triplicate recordings of the wedged hepatic venous pressure (WHVP) and free hepatic venous pressure (FHVP). The difference between FHVP and WHVP is referred to as HVPG, with values &#8805;10 mm Hg indicating clinically significant portal hypertension (CSPH). Before removing the catheter, pressure readings obtained in the IVC at the same level, as well as the right atrial pressure are recorded.
Finally, a transjugular liver biopsy can be obtained via the same vascular route. Different systems are available; however, core biopsy needles are preferred over aspiration needles, especially for cirrhotic livers. Again, under fluoroscopic guidance a biopsy needle introducer sheath is advanced into an hepatic vein. Next, the transjugular biopsy needle is gently advanced through the introducer sheath: (i) in case of aspiration biopsy, the needle is advanced into the liver parenchyma under aspiration and then removed quickly, or (ii) in case of a core biopsy, the cutting-mechanism is triggered inside the parenchyma. Several separate passages can be safely performed to obtain sufficient liver specimens via transjugular biopsy. In experienced hands, the combination of these procedures takes about 30-45 min.

Here, we present a protocol for measurement of hepatic venous pressure gradient (HVPG),the gold standard to diagnose clinically significant portal hypertension. Moreover, we describe how to perform a transjugular liver biopsy within the same session.

肝静脉压梯度和转性肝活检测量

Here we provide a detailed protocol describing the clinical procedure of hepatic venous pressure gradient (HVPG) measurement in patients with advanced ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

小鼠右心室系统及缺氧诱发肺动脉高血压的侵入性血液动力学评估

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

Evaluation of Cerebral Blood Flow Autoregulation in the Rat Using Laser Doppler Flowmetry

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained using laser Doppler flowmetry (LDF). This paper demonstrates a closed skull preparation that allows cerebral blood flow to be assessed without penetrating the skull or installing a chamber or cerebral window. To evaluate autoregulatory mechanisms, a model of controlled blood pressure reduction via graded hemorrhage can be utilized while simultaneously employing LDF. This enables the real time tracking of the relative changes in the blood flow in response to reductions in arterial blood pressure produced by the withdrawal of circulating blood volume. This paradigm is a valuable approach to study cerebral blood flow autoregulation during reductions in arterial blood pressure and, with minor modifications in the protocol, is also valuable as an experimental model of hemorrhagic shock. In addition to evaluating autoregulatory responses, LDF can be used to monitor the cortical blood flow when investigating metabolic, myogenic, endothelial, humoral, or neural mechanisms that regulate cerebral blood flow and the impact of various experimental interventions and pathological conditions on cerebral blood flow.

This article demonstrates the use of laser Doppler flowmetry to evaluate the ability of the cerebral circulation to autoregulate its blood flow during reductions in arterial blood pressure.

使用激光多普勒流测量测量大鼠脑血流自动调节的评估

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...