胃内容物的气管吸入模型

Bruce  A.  Davidson; Ravi  Alluri

doi:10.21769/BioProtoc.968

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
CN
- EN - English
- CN - 中文

Peer-reviewed

Gastric Aspiration Models

胃内容物的气管吸入模型

Bruce A. Davidson email

RA Ravi Alluri

发布: 2013年11月20日第3卷第22期 DOI: 10.21769/BioProtoc.968 浏览次数: 19609

评审: Anonymous reviewer(s)

下载 PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Immunology, featuring study using the protocol.

Feb 2013

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

基于参与者来源的异种移植小鼠模型：解析HIV自体控制机制并评估免疫疗法

Emma Falling Iversen [...] R. Brad Jones

2025年04月05日 1318 阅读

改良版Miles法分析血管通透性

Hilda Vargas-Robles [...] Michael Schnoor

2025年04月05日 1227 阅读

用于多发性硬化研究的PBMC人源化小鼠模型：聚焦免疫变化与中枢神经系统受累

Anastasia Dagkonaki [...] Lesley Probert

2025年05月20日 2102 阅读

Abstract

The procedures described below are for producing gastric aspiration pneumonitis in mice with alterations for rats and rabbits described parenthetically. We use 4 different injury vehicles delivered intratracheally to investigate the inflammatory responses to gastric aspiration:
1) Normal saline (NS) as the injury vehicle control
2) NS + HCl, pH = 1.25 (acid)
3) NS + gastric particles, pH ≈ 5.3 (part.)
4) NS + gastric particles + HCl, pH = 1.25 (acid + part.)
The volume, pH, and gastric particle concentration all affect the resulting lung injury. In mice, we generally use an injury volume of 3.6 ml/kg (rat: 1.2 ml/kg, rabbit: 2.4 ml/kg), an injury pH (for the acid-containing vehicles) of 1.25, and a gastric particulate concentration (in the particulate-containing vehicles) of 10 mg/ml (rat: 40 mg/ml). In our hands this results in a maximal, non-lethal lung injury with ≤ 10% mortality for the most injurious vehicle (i.e., acid + part.) The maximum tolerable particulate concentration needs to be determined empirically for any new strains to be used, especially in genetically-altered mice, because an altered inflammatory response may have detrimental affects on mortality.
We have extensive experience utilizing these procedures in the outbred strain, CD-1, as well as many genetically-altered inbred stains on the C57BL/6 background. Choice of strain should be carefully considered, especially in terms of strain-specific immune bias, to assure proper data interpretation. The size of the mouse should be ≥ 20 g at the time of injury. Smaller mice can be attempted, if necessary, but the surgical manipulation becomes increasingly more difficult and the surgery survival rate decreases substantially. There are no size or strain constraints for rat and rabbit models, but we generally use Long-Evans rats at 250-300 g and New Zealand White rats at ≈ 2 kg at the time of initial injury.

Keywords: Pneumonitis (肺炎)

Acute lung injury (ALI) (急性肺损伤（阿里）)

Acute respiratory distress syndrome (ARDS) (急性呼吸窘迫综合征（ARDS）)

Inflammation (炎症)

Rodent models (啮齿类动物模型)

Materials and Reagents

Isoflurane
Topical antiseptic microbicide prep solution (e.g. Medline Industries, catalog number: MDS093906 )
0.5% bupivicaine
Hank’s Balanced Salt Solution (HBSS) with Ca²⁺, Mg²⁺ (e.g. Life Technologies, catalog number: 14025 )
HBSS without Ca²⁺, Mg²⁺ (e.g. Life Technologies, catalog number: 14175 )
Liquid nitrogen
Bovine serum albumin (BSA)
Cytospin filter cards (e.g. Thermo Fisher Scientific, catalog number: 5991022 )
Diff Quik solutions kit (Fisher Scientific, catalog number: NC9943455 )
Cytoseal 60 (VWR International, catalog number: 48212-154 )
100x protease inhibitor cocktail (e.g. Calbiochem^®, catalog number: 80053-844 )
Bupivacaine
100 mg/ml mouse (rat) gastric particles (see Recipes)
Acid injury solution (acid) (see Recipes)
Gastric particles injury solution (part.) (see Recipes)
Acid + particles injury solution (acid + part.) (see Recipes)
Phosphate buffered saline (PBS), pH 7.2 (see Recipes)
Ammonium chloride lysis buffer (see Recipes)
Lung homogenate buffer (see Recipes)
50 mM potassium phosphate buffer, pH = 6.0 (see Recipes)
MPO homogenate buffer (see Recipes)

Note: All salts and other chemicals are from Sigma-Aldrich unless otherwise noted (however, any source for such chemicals is probably okay to use).

Equipment

1-0 braided silk material, bulk spool (e.g. Look catalog number: MBJF210)
12 mm x 75 mm x 1 mm microscope slides
2 x 2 gauze sponges, 8-ply (VWR International, catalog number: 82004-740 )
Sterile 4 x 4 gauze sponges, 8-ply (VWR International, catalog number: 82004-742 )
6-0 monofilament polypropylene suture with P-13 cutting needle (e.g. Syneture, catalog number: SP-5695 )
60° incline dissection board (homemade out of plexiglass)
Syringes (0.5, 1, 3, 5, 20 cc (cubic centimeter))
Needles (14, 20, 22, 26, 29 gauge)
Tracheal cannula (23 gauge x 1/2” stainless steel tubing adapter) (Becton, Dickinson and Company, catalog number: 408213 )
3” curved serrated forceps (2)
3” curved tissue (“toothed”) forceps
1.8 ml microfuge tubes
12 x 75 mm polystyrene tubes
22 x 22 mm #1.5 coverslips
4” curved micro dissecting scissors
37 °C water bath
Disposable skin stapler (e.g. 3M, model: DS-25 )
Hemocytometer or Coulter counter (e.g. Beckman Coulter, model: MultiSizer III )
Cytocentrifuge with cytospin funnels (e.g. Shandon CytoSpin^®)
Tissue homogenizer (e.g. Brinkmann^TMPolytron^TM, model: PT2000 )
Probe sonicator (e.g. Branson Sonifier^®, model: 450 )