Name: open tracheostomy gastric acid aspiration murine model of acute lung injury results in maximal acute nonlethal lung injury
Uploaded: 2017-02-26T14:00:00
Description: Watch this Scientific Journal Video about Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury at JoVE.com

Ravi Alluri and Hilliard L. Kutscher are supported by Ruth L. Kirschstein National Research Service Award (NRSA) Institutional Research Training Grant 1T32GM099607.

所有的材料和设备需要收集和适当前程序组织的。该过程是要以无缝转换从一个步骤到下一个，以便提供一致的，可重现的数据进行的。该协议遵循的机构动物护理和使用委员会在水牛城退伍军人医疗中心设置机构政策。 注意：两个雄性和雌性C57BL / 6小鼠用于这个协议。有两性之间白蛋白漏出无显著差异。 1.盐酸准备<ol><li>使56.2毫盐酸无菌加入1N盐酸281微升至4.72毫升的无菌生...

<ol>
	<li>Knight, P. R., Rutter, T., Tait, A. R., Coleman, E., Johnson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+gastric+particulate+lung+injury:+a+comparison+and+interaction+with+acidic+pneumonitis.">Pathogenesis of gastric particulate lung injury: a comparison and interaction with acidic pneumonitis.</a> Anesth Analg. 77 (4), 754-760 (1993).</li><li>Raghavendran, K., Nemzek, J., Napolitano, L. M., Knight, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aspiration-induced+lung+injury.">Aspiration-induced lung injury.</a> Crit Care Med. 39 (4), 818-826 (2011).</li><li>Rubenfeld, G. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+and+outcomes+of+acute+lung+injury.">Incidence and outcomes of acute lung injury.</a> N Engl J Med. 353 (16), 1685-1693 (2005).</li><li>Kennedy, T. 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=Acute+acid+aspiration+lung+injury+in+the+rat:+biphasic+pathogenesis.">Acute acid aspiration lung injury in the rat: biphasic pathogenesis.</a> Anesthesia &amp; Analgesia. 69 (1), 87-92 (1989).</li><li>Kudoh, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+pentoxifylline+on+acid-induced+alveolar+epithelial+injury.">The effect of pentoxifylline on acid-induced alveolar epithelial injury.</a> Anesthesiology. 82 (2), 531-541 (1995).</li><li>Prophet, E. B., Mills, B., Arrington, J. B., Sobin, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Laboratory methods in histotechnology. , (1995).</li><li>Kyriakides, 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=Membrane+attack+complex+of+complement+and+neutrophils+mediate+the+injury+of+acid+aspiration.">Membrane attack complex of complement and neutrophils mediate the injury of acid aspiration.</a> J Appl Physiol. 87 (6), 2357-2361 (1999).</li><li>Yoshikawa, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+ventilator-induced+vascular+permeability+and+cytokine+responses+in+isolated+and+in+situ+mouse+lungs.">Acute ventilator-induced vascular permeability and cytokine responses in isolated and in situ mouse lungs.</a> J Appl Physiol. 97 (6), 2190-2199 (2004).</li><li>Eyal, F. G., Hamm, C. R., Coker-Flowers, P., Stober, M., Parker, 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=The+neutralization+of+alveolar+macrophages+reduces+barotrauma-induced+lung+injury.">The neutralization of alveolar macrophages reduces barotrauma-induced lung injury.</a> FASEB J. 16 (4), 410-411 (2002).</li><li>Hermans, C., Bernard, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+epithelium-specific+proteins:+characteristics+and+potential+applications+as+markers.">Lung epithelium-specific proteins: characteristics and potential applications as markers.</a> Am J Respir Crit Care Med. 159 (2), 646-678 (1999).</li><li>Segal, B. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acid+aspiration-induced+lung+inflammation+and+injury+are+exacerbated+in+NADPH+oxidase-deficient+mice.">Acid aspiration-induced lung inflammation and injury are exacerbated in NADPH oxidase-deficient mice.</a> Am J Physiol Lung Cell Mol Physiol. 292 (3), 760-768 (2007).</li><li>Matthay, M. A., Robriquet, L., Fang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alveolar+epithelium:+role+in+lung+fluid+balance+and+acute+lung+injury.">Alveolar epithelium: role in lung fluid balance and acute lung injury.</a> Proc Am Thorac Soc. 2 (3), 206-213 (2005).</li><li>Richard, J. C., Factor, P., Welch, L. C., Schuster, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+the+spatial+distribution+of+transgene+expression+in+the+lungs+with+positron+emission+tomography.">Imaging the spatial distribution of transgene expression in the lungs with positron emission tomography.</a> Gene Ther. 10 (25), 2074-2080 (2003).</li></ol>

<html>其目的是通过胃酸的愿望是非常类似于酸肺炎和急性呼吸窘迫综合征后续开发过程中发生在人类的病理生理发展的ALI动物模型。在开发的模型，我们选择了提供高吞吐量数据采集动物物种，由于其低成本，短生殖周期，并用大量的调查工具（ 即 ，单克隆抗体，转基因的菌株）一个很好的理解免疫系统。 ALI继人类尖刻地损害肺泡上皮细胞和血管内皮屏障功能产生?...

急性呼吸窘迫综合征的特点是广泛的肺部炎症和临床上被视为呼吸短促急性低氧血症。这些症状通常是煽动事件发生后不到24小时，如外伤，败血症，输血相关反应或愿望。它被定位于肺泡上皮细胞和血管内皮细胞，导致蛋白质漏出，随后透明膜形成中性粒细胞肺泡炎（ 即广泛的炎症）组织病理学特点。吸入被归类为化学性肺炎或吸入性肺炎。 1胃吸出的酸性成分有助于肺炎和偏好都制定了继发性细菌性肺炎。吸入性肺炎是ALI和ARDS的后续发展的主要风险因素之一。 2 胃愿望是定义为从T物料吸入急性事件他带或不带口咽部菌群到气管超出声带胃。在抽吸内容物可以包含低pH的胃流体，细菌，血液或食品颗粒。胃吸出患者常发生在重症监护病房（ICU），其中通常是在空腹状态，因此放置在一个质子泵抑制剂限制酸化胃内容物误吸。比起一般患者群高5倍 - 急性肺损伤在美国ICU的人口的发病率是2.5。 3遗憾的是，这些易患因素往往会导致胃内细菌过度生长的状态，可能会导致下面的一个愿望事件肺部更严重的后遗症，如胃的愿望是继发细菌性肺炎发展的一个独立危险因素（SBP） ，ALI和急性呼吸窘迫综合征。 胃抽吸具有两个主要部分组成：盐酸和胃内容物，其可以或可以不含有细菌或食物颗粒。在啮齿动物模型单独胃抽吸的酸成分产生一个初始炎症应答作为对气道上皮低pH的直接苛性损伤的结果。这之后是中性粒细胞浸润，并在4炎性反应 - 6小时。 4这两个因素最终导致的肺微血管完整性从而导致流体和蛋白质外渗进入肺泡和呼吸道破坏。要理解本病理生理学和进一步研究可能的治疗干预，开发和表征即阐明涉及的基本机制的动物模型是重要的。单独的酸性抽吸必须庞大或具有足够低的pH值以绕过呼吸树的缓冲能力和到达肺泡。如果这不发生，只发生瞬态上，传导气道损伤，从而不太可能导致ARDS的严重后遗症。 5 &lt;/ SUP&gt; 为了准确地与受伤的肺泡上皮仿真抽吸事件，绕过动物的天然防御很重要。利用小鼠模型的愿望，产生ALI模仿人类看到的胃酸的伤害，必须占气管支气管树的差异。开气管切开术，该方法利用绕过损伤重现ALI既生理学和组织学的方式鼠和人的呼吸的树木和模型之间的差异。从历史上看，气管内插管用于生成ALI，然而它被认为是困难的小鼠未经喉损伤来执行。因此，这种方法提供了已取得在多个研究人员和以最小的过程归因死亡率一致的结果的潜在替代。

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			<td>syringe, 1cc</td>
			<td>Becton Dickinson</td>
			<td>309628</td>
			<td></td>
		</tr>
		<tr>
			<td>syringe, 5cc</td>
			<td>Becton Dickinson</td>
			<td>309646</td>
			<td></td>
		</tr>
		<tr>
			<td>needle, 22 ga x 1 1/2&#34;</td>
			<td>Becton Dickinson</td>
			<td>305159</td>
			<td></td>
		</tr>
		<tr>
			<td>needle, 26 ga x 1 1/2&#34;</td>
			<td>Becton Dickinson</td>
			<td>305111</td>
			<td></td>
		</tr>
		<tr>
			<td>1-O Braided Silk Suture</td>
			<td>Harvard Apparatus</td>
			<td>517730</td>
			<td></td>
		</tr>
		<tr>
			<td>3&#34; Curved tissue serrated forceps</td>
			<td>Fine Science Tools</td>
			<td>11065-07</td>
			<td></td>
		</tr>
		<tr>
			<td>3&#34; Curved tissue &#34;toothed&#34; forceps, 1x2 teeth</td>
			<td>Fine Science Tools</td>
			<td>11067-07</td>
			<td></td>
		</tr>
		<tr>
			<td>4&#34; curved micro dissecting scissors</td>
			<td>Fine Science Tools</td>
			<td>14061-10</td>
			<td></td>
		</tr>
		<tr>
			<td>bone cutting spring scissors</td>
			<td>Fine Science Tools</td>
			<td>16144-13</td>
			<td></td>
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			<td>3 1/2&#34; curved locking hemostat</td>
			<td>Fine Science Tools</td>
			<td>13021-12</td>
			<td></td>
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			<td>Disposable Skin Stapler</td>
			<td>3M</td>
			<td>DS-25</td>
			<td></td>
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			<td>tracheal cannula (20 ga x 1/2&#34; stainless steal tubing adapter)</td>
			<td>Becton Dickinson</td>
			<td>408210</td>
			<td></td>
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			<td>60-degree Incline Dissection Board</td>
			<td></td>
			<td></td>
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			<td>0.5% Bupivacaine</td>
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			<td></td>
			<td></td>
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			<td>Isoflurane</td>
			<td></td>
			<td></td>
			<td></td>
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			<td>Betadine and &#34;Q-tip&#34; cotton applicator</td>
			<td></td>
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open tracheostomy gastric acid aspiration murine model of acute lung injury results in maximal acute nonlethal lung injury

酸肺炎是无菌的急性肺损伤（ALI）的主要原因在人类。酸肺炎跨越临床谱从无症状到急性呼吸窘迫综合征（ARDS），其特征在于嗜中性肺泡炎，和伤害到两个肺泡上皮和血管内皮。在临床上，急性呼吸窘迫综合征是由低氧血症急性起病，双侧斑片状肺浸润和非心源性肺水肿定义。人类研究为我们提供了有关引起ARDS肺中的生理和炎性变化，这导致了对下属机制各种假说有价值的信息。不幸的是，在确定ARDS的病因，以及广泛的病理生理学的困难导致缺乏可能是在开发治疗策略有用的关键信息。 平移动物模型是有价值的，当他们发病机制和病理生理学准确reproducEA概念在体外和临床证实。虽然大动物模型（ 如羊）人的气管，支气管树的解剖结构的共同特征，小鼠模型提供的其他优势，包括主机：成本低;繁殖周期短借贷本身更大的数据采集;一个很好的理解免疫系统;和充分表征的基因组，导致的各种基因缺失和转基因的菌株的可用性。低pH诱导的急性呼吸窘迫综合征的鲁棒模型需要鼠ALI靶向主要肺泡上皮，其次的血管内皮，以及小气道导致肺泡。此外，不同的伤害性和非伤害性的侮辱之间的巨大差异可重复的伤害是很重要的。 用盐酸这里介绍的鼠胃酸误吸模型采用开放式气管切开术，并再现了致病场景，再现低pH pneumonITIS在人体的伤害。此外，该模型可用于研究低pH损伤与其它肺损害的实体（ 例如 ，食物颗粒，致病菌）的相互作用。

酸吸入性肺炎的小鼠模型的肺组织病理如上所述，并取出肺24小时后的低pH侮辱，切片，H＆E染色（ 图1）小鼠受伤。坏死细胞，肺实质结构，细胞和碎片空域内和显著PMN浸润的损失被清楚地观察到。类似于临床心愿，我们在产生斑片状浸润中性粒细胞和纤维蛋白蛋白沉积的区域远端肺空气空间的异质性损伤模型的结?...

Watch this Scientific Journal Video about Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury at JoVE.com

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

急性肺损伤开放气管切开胃酸吸入小鼠模型结果在急性最大非致命性肺损伤

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Research

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Medicine

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

压力控制通气诱导小鼠急性肺损伤

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

科研

医学

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

在背皮褶商会鼠标的缺血性组织损伤：一个皮瓣模型，探讨急性缺血持续

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Murine Model of Intestinal Ischemia-reperfusion Injury

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Oleic Acid-Injection in Pigs As a Model for Acute Respiratory Distress Syndrome

油酸注射液在猪急性呼吸窘迫综合征模型中的研究

The acute respiratory distress syndrome is a relevant intensive care disease with an incidence ranging between 2.2% and 19% of intensive care unit ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

慢性血栓栓塞性肺动脉高压大型动物模型中急性右心衰竭的诱导和表型分析

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

通过直接内切内脂多糖灌输诱导小鼠急性肺损伤

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...