Fig 2: The impact of sevoflurane on heme oxygenase-1 (HO-1) modulation during acute pulmonary inflammation. (A) Gene, and (B) protein expression of HO-1 was evaluated in the pulmonary tissue three and 24 h after lipopolysaccharide (LPS) exposure. The effects of sevoflurane on HO-1 gene expression were examined by real-time polymerase chain reaction (RT-PCR). HO-1 protein expression (green) in the lung tissue of wild-type animals was evaluated by immunofluorescence staining at indicated conditions (original magnification ×63; one representative image of three independent experiments is shown; n = 3). DAPI (blue) was used as nuclear marker. Mean fluorescence intensity (MFI) of HO-1 with or without LPS exposure was measured by using ImageJ (n = 6–8). (C) Adenosine receptor A2B (Adora2b) gene expression was determined in the lung tissue after sevoflurane treatment, respectively, sevoflurane and hemin combination three hours after LPS exposure (n = 6–8). (D) Immunofluorescence staining of Adora2b surface expression (green) in lung tissue of wild-type animals was evaluated at indicated conditions (original magnification ×63; one representative image of three independent experiments is shown; n = 3). DAPI (blue) was used as a nuclear marker. MFI of Adora2b surface expression was measured at indicated conditions by using ImageJ (n = 6–8). (E) The impact of HO-1 activation or inhibition on sevoflurane effects on polymorphonuclear neutrophil (PMN) migration 24 h after lipopolysaccharide (LPS) stimulation was detected (n = 8). (F) Surface expression of CD11a/CD18 (also known as lymphocyte function-associated antigen-1; LFA-1), and (G) CD162 (also called P-selectin glycoprotein 1; PSGL1) was evaluated on human PMNs after LPS exposure. The impact of sevoflurane and heme oxygenase modulation on the presence of CD11a/CD18 and CD162 (mean fluorescence intensity; MFI) on human PMNs was assessed by flow cytometry (n = 6–8). Statistical analyses were performed by one-way ANOVA + Bonferroni correction or Kruskal-Wallis test. Data are presented as mean ± SEM; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.