Sevoflurane suppresses tumour necrosis factor-α-induced inflammatory responses in small airway epithelial cells after anoxia/reoxygenation K. Watanabe,

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Sevoflurane suppresses tumour necrosis factor-α-induced inflammatory responses in small airway epithelial cells after anoxia/reoxygenation K. Watanabe, C. Iwahara, H. Nakayama, K. Iwabuchi, T. Matsukawa, K. Yokoyama, K. Yamaguchi, Y. Kamiyama, E. Inada British Journal of Anaesthesia Volume 110, Issue 4, Pages 637-645 (April 2013) DOI: 10.1093/bja/aes469 Copyright © 2013 The Author(s) Terms and Conditions

Fig 1 Effects of sevoflurane on TNF-α-induced IL-6, IL-8, and MCP-1 gene expression in SAEC. SAEC were stimulated by TNF-α (0, 5, 10, 50 ng ml−1) under A/R conditions without (blue bar) or with 1.1 vol% (green bar) or 2.2 vol% (red bar) sevoflurane. Levels of IL-6 (a), IL-8 (b), and MCP-1 (c) gene expression were measured by quantitative real-time RT–PCR. The data were normalized relative to β-actin as an internal control and are presented as mean (sd); n=3 per group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 for the comparison between A/R and A/R+sevo. British Journal of Anaesthesia 2013 110, 637-645DOI: (10.1093/bja/aes469) Copyright © 2013 The Author(s) Terms and Conditions

Fig 2 Effects of sevoflurane on TNF-α (50 ng ml−1)-induced production of IL-6 and IL-8 in SAEC. Concentrations of IL-6 (a) and IL-8 (b) in cell supernatants were measured by ELISA. Data are presented as mean (sd); n=3 per group. ****P<0.0001 for the comparison between A/R and A/R+sevo. British Journal of Anaesthesia 2013 110, 637-645DOI: (10.1093/bja/aes469) Copyright © 2013 The Author(s) Terms and Conditions

Fig 3 Effect of sevoflurane on TNF-a-induced activation of p65 NF-κB. (a) The effect of sevoflurane on nuclear translocation of p65 NF-κB was analysed by immunoblotting. (b) Band density was measured by densitometric analysis. Data were normalized relative to lamin C as an internal control, and are presented as mean (sd) of three independent experiments. **P < 0.01 for the comparison between A/R and A/R+sevo. (c–e) Levels of NFKB1, NFKBIA, and NFKBIE gene expression were measured by quantitative real-time RT-PCR. Data were normalized relative to β-actin as an internal control. Black and white bars represent the experiments without and with sevoflurane, respectively. Data are presented as mean (sd); n = 3 per group. *P < 0.05 for the comparison between A/R and A/R+sevo. British Journal of Anaesthesia 2013 110, 637-645DOI: (10.1093/bja/aes469) Copyright © 2013 The Author(s) Terms and Conditions

Fig 4 Effect of sevoflurane on mitochondrial integrity in TNF-α-stimulated SAEC. (a) SAEC were stained with JC-1 and analysed by flow cytometry. Ordinate: mitochondrial membrane potential (Δψm). CCCP, carbonyl cyanide 3-chlorophenylhydrazone (a reagent that causes rapid depolarization of Δψm). (b) To account for variations in mitochondrial volume, the ratio of FL2/FL1 shown in (a) was calculated. Each bar represents the mean (sd) of three experiments. British Journal of Anaesthesia 2013 110, 637-645DOI: (10.1093/bja/aes469) Copyright © 2013 The Author(s) Terms and Conditions