Α7 Nicotinic acetylcholine receptor agonist GTS-21 attenuates ventilator-induced tumour necrosis factor-α production and lung injury M. Kox, J.C. Pompe,

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α7 Nicotinic acetylcholine receptor agonist GTS-21 attenuates ventilator-induced tumour necrosis factor-α production and lung injury M. Kox, J.C. Pompe, E. Peters, M. Vaneker, J.W. van der Laak, J.G. van der Hoeven, G.J. Scheffer, C.W. Hoedemaekers, P. Pickkers British Journal of Anaesthesia Volume 107, Issue 4, Pages 559-566 (October 2011) DOI: 10.1093/bja/aer202 Copyright © 2011 The Author(s) Terms and Conditions

Fig 1 Inflammatory cytokine levels in plasma (a) and lung homogenate (b) of non-ventilated (NV, n=8), placebo-treated and ventilated (placebo–MV, n=18) and GTS-21-treated and ventilated (GTS21–MV, n=16) mice. Lung homogenate cytokine levels are normalized to the total amount of protein in each homogenate. ANOVA P-values: plasma: TNF-α P=0.0002, IL-6 P=0.0008, KC P=0.0035, IL-10 P<0.0001. Lung: TNF-α P=0.034, IL-6 P<0.0001, KC P=0.0007, IL-10 P=0.0048. Data are represented as mean (sd). *P<0.05 compared with placebo–MV (the Bonferroni post hoc test of placebo–MV vs GTS21–MV). British Journal of Anaesthesia 2011 107, 559-566DOI: (10.1093/bja/aer202) Copyright © 2011 The Author(s) Terms and Conditions

Fig 2 mRNA expression (relative to the housekeeping gene β-actin) of inflammatory cytokines in lung tissue of non-ventilated (NV, n=7), placebo-treated and ventilated (placebo–MV, n=7), and GTS-21 treated and ventilated (GTS21–MV, n=7). ANOVA P-values: TNF-α P=0.0017, IL-6 P=0.45, IL-10 P=0.0013. Horizontal line indicates mean. *P<0.05 compared with placebo–MV (the Bonferroni post hoc test of placebo–MV vs GTS21–MV). British Journal of Anaesthesia 2011 107, 559-566DOI: (10.1093/bja/aer202) Copyright © 2011 The Author(s) Terms and Conditions

Fig 3 TNF-α production (a), represented as per cent of LPS-induced production and mRNA expression (b), relative to the housekeeping gene β-actin) in isolated alveolar macrophages stimulated with LPS (100 ng ml−1) for 4 h in the presence and absence of GTS-21 (100 μM). TNF-α production in the absence of LPS is below the detection limit (32 pg ml−1); mean LPS-induced TNF-α production is 2503 (1462) pg ml−1. Data are represented as individual data points of three macrophage cultures obtained from three different mice. Horizontal lines indicate mean. British Journal of Anaesthesia 2011 107, 559-566DOI: (10.1093/bja/aer202) Copyright © 2011 The Author(s) Terms and Conditions

Fig 4 Inflammatory cytokine levels in plasma (a) and lung homogenate (b) of non-ventilated (NV, n=8), placebo-treated and ventilated (placebo–MV, n=15), neostigmine-treated and ventilated (neostigmine–MV, n=14), and mecamylamine-treated and ventilated (mecamylamine–MV, n=15) mice. Lung homogenate cytokine levels are normalized to the total amount of protein in each homogenate. ANOVA P-values: plasma: TNF-α P=0.0046, IL-6 P<0.0001, KC P=0.0051, IL-10 P=0.0060. Lung: TNF-α P=0.0462, IL-6 P=0.0250, KC P=0.0003, IL-10 P=0.5006. Data are represented as mean (sd). British Journal of Anaesthesia 2011 107, 559-566DOI: (10.1093/bja/aer202) Copyright © 2011 The Author(s) Terms and Conditions