Models of Inflammation: Carrageenan Air Pouch

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Abstract
Table of Contents
Materials
Figures
Literature Cited

Abstract

The subcutaneous air pouch is an in vivo model that can be used to study acute and chronic inflammation, the resolution of the inflammatory response, and the oxidative stress response. Injection of irritants into an air pouch in rats or mice induces an inflammatory response that can be quantified by the volume of exudate produced, the infiltration of cells, and the release of inflammatory mediators. The model presented in this unit has been extensively used to identify potential anti?inflammatory drugs. Curr. Protoc. Pharmacol. 56:5.6.1?5.6.8. © 2012 by John Wiley & Sons, Inc.

Keywords: inflammation; edema; carrageenan; air pouch; animal model; inflammatory mediator; PGE2; TNF alpha

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Introduction
Basic Protocol 1: Air Pouch Model in the Rat
Alternate Protocol 1: Air Pouch Model in the Mouse
Reagents and Solutions
Commentary
Literature Cited
Figures
Tables

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Materials

Basic Protocol 1: Air Pouch Model in the Rat

Materials

Rats weighing ∼150 g or mice weighing ∼20 to 25 g
Inhalation anesthetic [e.g., 5% (v/v) isoflurane; IsoFlo, Abbott]
70% (v/v) ethanol
0.9% (w/v) saline, sterile
2% carrageenan solution (see recipe )
Lavage solution (see recipe )
ELISA kit(s) specific for inflammatory mediators (e.g., TNF‐α, IL‐1β, IL‐6, IL‐10, prostaglandins; Thermo Scientific)
Isotonic phosphate‐buffered saline (PBS; Invitrogen, cat. no. 10010‐031), ice cold
Differential staining kit (e.g., Hemacolor)

5‐, 10‐, and 20‐ml syringes, sterile
18‐, 20‐ and 23‐G, 1‐ to 1.5‐in. needles, sterile
0.2‐µm syringe filters, sterile
Anesthetic chamber or chemical fume hood
Animal hair clippers
Hemostat and scissors
15‐ml conical centrifuge tubes, sterile
Transfer pipets, sterile
Cell counter or hemacytometer
Benchtop centrifuge (for 15‐ml conical tubes)

Additional reagents and equipment for euthanizing the animal (see Donovan and Brown, )

NOTE: Besides carrageenan, other irritants such as zymosan or lipopolysaccharide (LPS) can be used to induce inflammation in this model.CAUTION: Make certain that excess anesthetic does not vent into the room; use an appropriate anesthesia chamber apparatus or work in a chemical fume hood.

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Figures

Figure 5.6.1 Analysis of cellular infiltrate in rat air pouch exudates after carrageenan stimulation. Data for each time point represent (A ) mean number of cells and (B ) percent of total cells for each of four cell types in a group of eight rats for each time point. Data at 0 hr are for untreated animals.

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Figure 5.6.2 Amounts of tumor necrosis factor α (TNF‐α) and prostaglandin E₂ (PGE₂ ) present in rat air pouch exudates 3 hr after carrageenan stimulation. Symbols: filled diamond, ng/ml TNF‐α or PGE₂ detected for each rat in the experimental group; horizontal line, median level of production for the experimental group. Exudates from unstimulated air pouches contained no detectable levels of TNF‐α or PGE₂ . Note the extent of variation among the individuals in both groups.

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Videos

Literature Cited

Literature Cited
	Ariel, A. and Serhan, C.N. 2007. Resolvins ad protectins in the termination program of acute inflammation. Trends Immunol. 28:176–183.
	Donovan, J. and Brown, P. 2006. Euthanasia. Curr. Protoc. Immunol. 73:1.8.1‐1.8.4.
	Edwards, J.C.W., Sedgwick, A.D., and Willoughby, D.A. 1981. The formation of a structure with the features of synovial lining by subcutaneous injection of air: An in vivo tissue culture system. J. Pathol. 134:147–153.
	Jain, M. and Parmar, H.S. 2011. Evaluation of antioxidative and anti‐inflammatory potential of hesperidin and naringin on the rat air pouch model of inflammation. Inflamm. Res. 60:483–491.
	Martin, S.W., Stevens, A.J., Brennan, B.S., Davies, D., Rowland, M., and Houston, J.B. 1994. The six‐day‐old rat air pouch model of inflammation: characterization of the inflammatory response to carrageenan. J. Pharmacol. Toxicol. Methods 32:139–147.
	Masferrer, J.L., Zweifel, B.S., Manning, P.T., Hauser, S.D., Leahy, K.M., Smith, W.G., Isakson, P.C., and Seibert, K. 1994. Selective inhibition of inducible cyclooxygenase 2 in vivo is antiiflammatory and nonulcerogenic. Proc. Natl. Acad. Sci. U.S.A. 91:3228‐3232.
	Romano, M., Faggioni, R., Sironi, M., Sacco, S., Echtenacher, B., Di Santo, E., Salmona, M., and Ghezzi, P. 1997. Carrageenan‐induced acute inflammation in the mouse air pouch synovial model. Role of tumour necrosis factor. Mediators Inflamm. 6:32‐38.
	Sedgwick, A.D. and Lees, P. 1986. A comparison of air pouch, sponge and pleurisy models of acute carrageenan inflammation in the rat. Agents Actions 18:439‐446.
	Sedgwick, A.D., Koh, M.S., Willoughby, D.A., and Pelletier, M. 1981. Effects of sera and exudate from carrageenan‐treated rats on two models of acute inflammation. Agents Actions 11:477–481.
	Selye, H. 1953. On the mechanism through which hydrocortisone affects the resistance of tissue to injury. J. Am. Med. Assoc. 152:1207‐1213.
	Serhan, C.N., Krishnamoorthy, S., Recchiuti, A., and Chiang, N. 2011. Novel anti‐inflammatory–pro‐resolving mediators and their receptors. Curr. Top. Med. Chem. 11:629–647.
	Sin, Y.M., Sedgwick, A.D., Chea, E.P., and Willoughby, D.A. 1986. Mast cells in newly formed lining tissue during acute inflammation: a six day air pouch model in the mouse. Ann. Rheumatic Dis. 45:873–877.
	Vane, J.R., Mitchell, J.A., Appleton, I., Tomlinson, A., Bishop‐Bailey, D., Croxtall, J., and Willoughby, D.A. 1994. Inducible isoforms of cyclooxygenase and nitric‐oxide synthase in inflammation. Proc. Natl. Acad. Sci. U.S.A. 91:2046–2050.

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Models of Inflammation: Carrageenan Air Pouch

Abstract

Table of Contents

Materials

Basic Protocol 1: Air Pouch Model in the Rat

Figures

Videos

Literature Cited