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1 Department of Physiology, University of Bergen, Norway 2 Broegelmann Research Laboratory, The Gade Institute, University of Bergen, Norway 3 Department of Paediatrics, Haukeland University Hospital, Bergen, Norway
Tumour necrosis factor-
(TNF-) and interleukin-1ß (IL-1ß) are important mediators produced during inflammation. We hypothesized that the pro-inflammatory cytokine response in the interstitial fluid (IF) is different from that in serum, and we aimed at quantifying the amount of TNF- and IL-1ß in the IF. By centrifugation of rat skin at < 424 g pure IF is extracted. Using ELISA such fluid was analysed for cytokines in back and/or paw skin of pentobarbital-anaesthetized rats, after either induction of endotoxaemia or ischaemia–reperfusion (I/R) injury. During endotoxaemia, TNF- increased in the IF from 0 in control to 640 ± 100 pg ml–1 (mean ±S.E.M.) after 90 min, with the serum concentration being 5–10 times higher at all time points. The response pattern of IL-1ß after lipopolysaccharide (LPS) challenge differed greatly from that of TNF- with a large increase in IF from 390 ± 90 to 28 000 ± 1500 pg ml–1 after 210 min, and a significantly smaller increase in serum (600 ± 45 pg ml–1). During reperfusion of the hind paw after 2 h of ischaemia, there was a gradual increase of TNF- in both IF of the paw skin and serum after 3 min of reperfusion. Both declined after 20 min. The pattern for IL-1ß differed, increasing significantly less in serum (25 ± 15 pg ml–1 after 20 min of reperfusion) than in the IF (1100 ± 200 pg ml–1). Immunostaining of the inflamed tissues showed increased expression of the two cytokines in cells of both epidermis and dermis compared to controls. Subdermal injections of TNF- and IL-1ß at the same concentrations found in IF after LPS infusion affected interstitial fluid pressure significantly. Local TNF- production dominates after I/R injury, whereas in endotoxaemia systemic production predominates. For IL-1ß local production dominates in both conditions. Thus, there is a differential pattern of cytokine production and the current method allows the study of the role of cytokines in IF during different inflammatory reactions.
(Received 23 October 2003;
accepted after revision 8 January 2004;
first published online 14 January 2004)
Corresponding author T. Nedrebø: Department of Physiology, University of Bergen, Jonas Lies vei 91, N-5009 Bergen, Norway. Email: torbjorn.nedrebo{at}fys.uib.no
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