压力超载诱发右室肥厚和衰竭的肺动脉扣条柱模型
Summary
我们提出了一种手术方法, 以诱导右心室肥厚和失败的大鼠。
Abstract
持续压力过载引起的右心室衰竭是导致几种心肺疾病发病和死亡的主要原因。因此, 有必要建立可靠和可重复的 rv 故障动物模型, 以调查疾病机制和潜在治疗策略的效果。肺干带是一种常见的方法, 以诱导孤立的 rv 肥大, 但一般来说, 前面描述的模型没有成功地创建一个稳定的模型的 rv 肥大和失败。
我们提出了一个大鼠模型的压力超载诱导 rv 肥大引起的肺干带 (ptb), 使不同的表型的 rv 肥大与 rv 失败和没有。我们使用改良的结扎夹贴花将肺主干周围的钛夹压缩到预先设定的内径。我们使用不同的剪辑直径, 以诱导不同阶段的疾病进展, 从轻度 rv 肥大到失代偿 rv 失败。
rv 肥大在接受 ptb 手术的大鼠中持续发展, 根据应用带夹的直径, 我们可以准确地再现不同的疾病严重程度, 从补偿肥大到严重的失代偿性 rv 不等心脏外表现的失败。
所提出的 ptb 模型是压力过载诱导 rv 肥大和失效的有效而稳健的模型, 它对其他带状模型具有多大的优点, 包括高重现性和诱发严重和失代偿的 rv 故障的可能性。
Introduction
右心室 (rv) 可以适应持续的压力过载。然而, 随着时间的推移, 自适应机制无法维持心脏输出量, rv 扩张, 最终 rv 失效。rv 功能是几种心肺疾病的主要预后因素, 包括肺动脉高压 (pah)、血栓栓塞性肺动脉高压 (cteph) 和各种形式的先天性心脏病, 有压力 (或体积) 超载的 rv。尽管经过了密集的治疗, 但在这些情况下, rv 故障仍然是主要的死亡原因。
由于 rv 的独特特性1、2和胚胎发育3 , 从左心力衰竭中获得的知识不能简单地推断为右心力衰竭。因此, 右心力衰竭的动物模型是必要的, 以调查 rv 失败的机制和潜在的药理治疗策略。
有 su5416 合并缺氧 (suhx) 4 或单氯他林 (mct)5引起肺动脉高压衰竭的实验模型, 可引起肺血管疾病继发的 rv 衰竭。这些模型用于评估针对肺血管的药物的治疗效果。suhx 和 mct 模型都是 rv 失效的非固定后载模型。因此, 无法确定干预后 rv 功能的改善是否继发于后负荷减少肺血管效应, 或者是否由对 rv 的直接影响引起。此外, mct 模型有几个心脏外的影响。
在实验肺干带模型中, 由于肺干的机械收缩, 房车的后负荷是固定的。这允许调查干预对 rv 的直接心脏影响独立于任何肺血管效应6,7,8,9。通常, 条带是通过沿肺干针进行的。然后在针头和肺干周围放置一个结扎, 并用一个结捆绑, 针头被移除, 将缝合线留在肺干周围。根据针头的规格, 可以应用不同程度的收缩, 但尽管这种方法被广泛使用, 它有一些缺点。首先, 带的直径与针头的外径不完全一样, 因为结扎是绑在针和肺干上的。其次, 结的紧密程度可能会有显著的变化, 很难复制一定程度的条带。这将导致条带直径的变化, 从而更大的分散。最后, 结可能会随着时间的推移而松动。
一项研究应用了肺干10周围的半封闭钽夹。他们将肺干周围的夹子压缩到 1.10 mm 2 的内部区域,并将其与使用 18 g 针缝合的大鼠进行比较。总体而言, 与剪辑的带与较少的围手术并发症和数据差异。
基于 schou 等人11 所述的原则, 进一步发展和表征了 rv 肥大和衰竭的肺主干条带 (ptb) 模型。在这里, 我们根据以前的研究 12、13 的结果介绍我们使用这个模型的经验。对于这种模型, 钛夹被压缩在肺干周围到一个精确的预设内径, 这可以调整, 以诱导不同的 rv 失败表型。
Protocol
Representative Results
Discussion
我们描述了一种方便和高度重现的方法肺躯干条带使用修改结扎夹苹果压缩肺干圈周围的钛夹。通过调整应用程序将夹子压缩到不同的内径, 可以诱导不同的 rv 肥大表型和失败, 包括严重的 rv 衰竭和心脏外失代偿表现。
虽然很简单, 但该协议包含几个关键步骤。重要的是, 老鼠在接受 ptb 手术时不能太大。根据我们的经验, 体重100–120克的 wistar 大鼠断奶者适用于该程序。在较?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作得到了丹麦独立研究理事会 [11e108410]、丹麦心脏基金会 [12e04-r90-a3852 和 12e04-r90-a3907] 和诺沃诺盘基金会 [NNF16OC0023244] 的支持。
Materials
| 17G IV Venflon Cannula | Becton Dickinson, US | 393228 | Distal 2 mm of the needle have been cut off |
| 1 mL syringe + 26G needle | Becton Dickinson, US | 303172 & 303800 | |
| 4-0 absorbable multifilament suture | Covidien, US | GL-46-MG | Polysorb, violet, 5×18" |
| 4-0 multifilament ligature | Covidien, US | LL-221 | Polysorb, violet, 98" |
| Buprenorphine | Indivior UK Limited | Local procurement, Temgesic 0.3 mg/mL | |
| Carprofene | ScanVet, DK | 27693 | Norodyl 50 mg/mL |
| Chlorhexidine | Faaborg Pharma, DK | Local procurement | |
| Contractor | Aesculap, Germany | BV010R | Blunt, self retaining, 70 mm |
| Ear Hooklet | Lawton, Germany | 66-0261 | Small, 14 cm, tip modified to an angle of 85° |
| Eye gel | Decra, UK | Lubrithal, Local procurement | |
| Forceps, Delicate Tissue | Lawton, Germany | 09-0020 | |
| Forceps, Dissecting | Lawton, Germany | 09-0013 | 1 regular, 1 with tip modified to an angle of 100° |
| Gas Anesthesia System | Penlon Limited, UK | SD0217SL | Sigma Delta Vaporizer |
| Hair trimmer | Oster | 76998-320-051 | |
| Horizon Open Ligating Clip Applier | Teleflex, US | 137085 | Modified with adjustable stop mechanism |
| Horizon Titanium Clips | Teleflex, US | 001200 | Small |
| Induction chamber | N/A | ||
| Iris Scissor | Lawton, Germany | 05-1450 | |
| Iris Scissor | Aesculap, Germany | BC060R | |
| Mechanical ventilator | Ugo Basile, Italy | 7025 | |
| Microscissor | Lawton, Germany | 63-1406 | |
| Microscope | Carl Zeiss, Germany | 303294-9903 | |
| Needle Holder | Lawton, Germany | 08-0011 | TITEGRIP |
| Pean | Lawton, Germany | 06-0100 | Halsted-Mosquito, straight |
| Pro-Optha | Lohmann & Rauscher, Germany | 16515 | Tampon |
| Saline 9 mg/mL | Fresenius Kabi, DK | 209319 | |
| Sevoflurane | AbbVie, US | Sevorane, Local procurement | |
| Surgical hook | Lawton, Germany | 51-0665 | Cushing, 19 cm, tip modified to an angle of 90° |
| Surgical Tape | 3M, US | 1530-0 | Micropore |
| Temperature Controller | CMA Microdialysis; Sweden | 8003760 | CMA 450 |
| Weighing machine | VWR, US | ||
| Wistar rat weanlings | Janvier Labs, France | RjHan:WI, 100-120 g |
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