无输血或强心剂剂的大鼠心肺旁路移植模型
Summary
在这里, 我们提出一个协议来描述一个简单的恢复心肺旁路模型没有输血或强心剂剂的大鼠。该模型可用于体外循环术后长期多脏器后遗症的研究。
Abstract
体外循环 (CPB) 在心血管手术中是必不可少的。尽管体外循环技术和设备的戏剧性改良, 多脏器并发症与长期体外循环仍然影响心血管手术的结果, 并可能恶化的术后发病率和死亡率。动物模型综述的临床应用可以澄清在体外循环期间发生的病理生理过程, 并促进临床前期研究, 以制定预防这些并发症的战略。大鼠体外循环模型是有利的, 因为他们更大的成本效益, 方便的实验过程, 丰富的测试方法在基因或蛋白质水平, 和遗传一致性。它们可用于调查免疫系统激活和促炎性细胞因子的合成, 赞美活化和氧自由基的产生。大鼠模型已被提炼, 并逐渐取代了大型动物模型。在这里, 我们描述一个简单的体外循环模型, 没有输血和/或强心剂剂的老鼠。这种恢复模式允许研究体外循环的长期多器官后遗症。
Introduction
在 1953年, 约翰·小长臂猿成功地使用了 CPB1进行了第一次心脏手术, 随后它成为心血管外科的基本形态。虽然这些技术和设备已大为完善, 但与体外循环有关的多脏器并发症仍会损害心血管手术的结果, 并可能影响术后发病率和死亡率2。与体外循环有关的器官损害是由免疫系统活化和促炎细胞因子的合成、赞美活化和氧自由基的产生而引起的2。然而, 它的病理生理学尚未完全阐明。
动物模型综述的临床应用, 使对体外循环期间和之后的病理生理过程的澄清;这可以促进临床前研究, 以制定战略, 以避免这些并发症。自从鲍佩维奇et al。第一次报告了鼠体外循环模型在 1967年3, 大鼠体外循环模型已经完善, 并已逐渐取代大型动物模型, 由于更大的成本效益, 方便的实验过程, 和大量的测试方法在遗传和蛋白质水平。此外, 近亲繁殖的大鼠可以遗传相同, 减少可能的生物偏见。
Fabre et al。首先建立了恢复模型, 允许对 CPB 的长期多器官后遗症进行研究4。这种简单的生存模式的优点是灵活性 (体外循环流量和持续时间), 稳定的生命条件, 和重现性的系统性炎症。大鼠体外循环模型已成为研究预防体外循环5中多脏器损伤的治疗策略的关键, 最近开发了各种模拟体外循环临床情况的模型。De 兰格et al。建立了心脏骤停模型, 可用于描述与心肌损伤有关的酶、遗传和组织学反应7。彼得斯et al。采用小型化体外循环模型进行心肌梗死和控制再灌注, 通过局灶性缺血再灌注损伤分析心脏功能障碍8。Jungwirth et al。首先建立了深低温循环骤停 (DHCA) 模型, 它可以阐明 DHCA 的全球缺血再灌注损伤, 支持潜在神经保护策略6。研究使用 DHCA 调查体温过低, 再灌注和/或溶血触发信号事件的影响9。深低温可能会影响各种酶和通路的活化和失活, 而机制仍然未知的10。另一方面, 心脏骤停模型或心脏缺血模型必须用于研究缺血再灌注心脏损伤。这些不同的大鼠体外循环模型, 高度重述人类体外循环可能揭示与体外循环有关的病理过程, 并有助于缓解与体外循环相关的并发症。
该协议说明了一个简单的体外循环模型, 没有输血或强心剂剂的大鼠。该模型可用于研究体外循环的长期多脏器后遗症。
Protocol
Representative Results
Discussion
在大鼠体外循环模型中, 体外循环后, 炎症细胞因子和 HMGB-1 的血清和肺表达水平是调节炎症反应的关键转录因子, 显著增加。以往的临床研究表明, 在心血管手术患者中, HMGB-1 水平的血清分泌增加了11, 体外循环期间血清 HMGB-1 水平与更严重的全身炎症反应综合征有关。体外循环后肺氧合功能障碍12。此外, 血清 HMGB-1 水平是一种独立的生物标志物, 用于预测重症肺?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
对塔基博士和 m. Funamoto 博士的技术支持给予了赞赏。
Materials
| Rodent Ventilator 7025 | Ugo Basile | 7025 | Ventilator |
| OxiQuant B | ENVITEC | 46-00-0023 | Oxygen Sensor |
| CMA 450 Temperature Controller | CMA | 8003759 | Temperature Controller |
| CMA 450 Heating Pad | CMA | 8003763 | |
| CMA 450 Rectal Probe | CMA | 8003761 | |
| DIN(8) to Disposable BP Transducer | ADInstruments | MLAC06 | |
| Disposable BP Transducer | ADInstruments | MLT0670 | |
| IX-214 Data Recorder | iWorx Systems | IWX-214 | amplifier |
| LabScribe software | iWorx Systems | software | |
| Roller pump | Furue Science | Model RP-VT | pump |
| Happy Cath | Medikit | EB 19G 4HCLs PP | 17-gauge multiorifice angiocatheter |
| SURFLO ETFE I.V. Catheter | Terumo | SR-OX2419CA | 24-gauge angiocatheter |
| Oxygenator | Mera | HPO-002 | |
| CPB circuit | Mera | custom-made | |
| Hespander fluid solution | Fresenius Kabi | 3319547A4035 | Hydroxyethyl starch |
Riferimenti
- Gibbon, J. H. Application of a mechanical heart and lung apparatus to cardiac surgery. Minn Med. 37 (3), 171-185 (1954).
- Apostolakis, E., Filos, K. S., Koletsis, E., Dougenis, D. Lung dysfunction following cardiopulmonary bypass. J Cardiac Surg. 25 (1), 47-55 (2010).
- Popovic, P., Horecky, J., Popovic, V. P. Instrumental responses in rats after hypothermic cardiopulmonary by-pass. P Soc Exp Biol Med. 126 (1), 225-228 (1967).
- Fabre, O., et al. A recovery model of partial cardiopulmonary bypass in the rat. Perfusion. 16 (3), 215-220 (2001).
- Hirao, S., Masumoto, H., Minatoya, K. Rat cardiopulmonary bypass models to Investigate multi-organ injury. Clin Surg. 2, 1-6 (2017).
- Jungwirth, B., et al. Neurologic outcome after cardiopulmonary bypass with deep hypothermic circulatory arrest in rats: description of a new model. J Thorac Cardiov Sur. 131 (4), 805-812 (2006).
- de Lange, F., Yoshitani, K., Podgoreanu, M. V., Grocott, H. P., Mackensen, G. B. A novel survival model of cardioplegic arrest and cardiopulmonary bypass in rats: a methodology paper. J Cardiothorac Surg. 3, 51 (2008).
- Peters, S., et al. An experimental model of myocardial infarction and controlled reperfusion using a miniaturized cardiopulmonary bypass in rats. Interact Cardiovasc Th. 19 (4), 561-564 (2014).
- Engels, M., et al. A cardiopulmonary bypass with deep hypothermic circulatory arrest rat model for the investigation of the systemic inflammation response and induced organ damage. J Inflamm. 11 (26), (2014).
- Pinto, A., et al. The extracellular isoform of superoxide dismutase has a significant impact on cardiovascular ischaemia and reperfusion injury during cardiopulmonary bypass. Eur J Cardio-Thorac. 50 (6), 1035-1044 (2016).
- Zhang, Z., Wu, Y., Zhao, Y., Xiao, X., Liu, J., Zhou, X. Dynamic changes in HMGB1 levels correlate with inflammatory responses during cardiopulmonary bypass. Exp Ther Med. 5 (5), 1523-1527 (2013).
- Kohno, T., et al. Impact of serum high-mobility group box 1 protein elevation on oxygenation impairment after thoracic aortic aneurysm repair. Heart Vessels. 26 (3), 306-312 (2011).
- Tseng, C. C., et al. Impact of serum biomarkers and clinical factors on intensive care unit mortality and 6-month outcome in relatively healthy patients with severe pneumonia and acute respiratory distress syndrome. Dis Markers. 2014, (2014).
- Paparella, D., Yau, T. M., Young, E. Cardiopulmonary bypass induced inflammation: pathophysiology and treatment. An update. Eur J Cardio-Thorac. 21 (2), 232-244 (2002).
- Hirao, S., et al. Recombinant human soluble thrombomodulin prevents acute lung injury in a rat cardiopulmonary bypass model. J Thorac Cardiov Sur. , (2017).
- Yamazaki, S., Inamori, S., Nakatani, T., Suga, M. Activated protein C attenuates cardiopulmonary bypass-induced acute lung injury through the regulation of neutrophil activation. J Thorac Cardiov Sur. 141 (5), 1246-1252 (2011).
- Wang, C. T., Zhang, L., Wu, H. W., Wei, L., Xu, B., Li, D. M. Doxycycline attenuates acute lung injury following cardiopulmonary bypass: involvement of matrix metalloproteinases. Int J Clin Exp Patho. 7 (11), 7460-7468 (2014).
- Liu, K., et al. Curcumin attenuates cardiopulmonary bypass-induced lung oxidative damage in rats. J Cardiovasc Pharm T. 17 (4), 395-402 (2012).
- Taki, T., et al. Fetal mesenchymal stem cells ameliorate acute lung injury in a rat cardiopulmonary bypass model. J Thorac Cardiov S. 153 (3), 726-734 (2017).
- Zhu, X., et al. Establishment of a novel rat model without blood priming during normothermic cardiopulmonary bypass. Perfusion. 29, 63-69 (2014).
- Inoue, K., et al. Deep anesthesia worsens outcome of rats with inflammatory responses. Inflamm Res. 65 (7), 563-571 (2016).
- Bradfield, J. F., Schachtman, T. R., McLaughlin, R. M., Steffen, E. K. Behavioral and physiologic effects of inapparent wound infection in rats. Lab Anim Sci. 42 (6), 572-578 (1992).
