Anti-Inflammatory Effects of Hyperbaric Oxygenation during DSS-Induced Colitis in BALB/c Mice Include Changes in Gene Expression of HIF-1α, Proinflammatory Cytokines, and Antioxidative Enzymes

Figure 2

HBO₂ changes immune cell frequencies in the mesenteric lymph nodes (MLN) and the spleen of BALB/c mice with DSS induced colitis. BALB/c mice at the age of 10–12 weeks were randomly assigned into 4 groups ( min. 5 mice/group/experiment): CTRL: control mice, CTRL + HBO₂: control mice undergoing HBO₂ (60 min/2.4 ATM, 2x/day, days 1–8), DSS: mice receiving dextran sodium sulphate (DSS, 5% w/v, days 1–7), and DSS + HBO₂: DSS treated mice undergoing HBO₂. Representative dot plots of MLN obtained by flow cytometry, illustrating the gating strategy for CD45⁺ Gr-1⁺ and CD45⁺ F4/80⁺ cells in MLN of CTRL, DSS, and DSS + HBO₂ groups. Doublets were excluded by forward scatter area (FSC-A) versus forward scatter height (FSC-H) and the dead cells using 7-AAD. (a) Frequency of CD45⁺ Gr-1⁺ cells, CD45⁺ F4/80⁺ cells, CD3⁺ T cells, and B220⁺ B cells in MLN (b) and spleen (c). Data are presented as mean s.e.m.% of single, live leukocytes (for Gr-1⁺ and F4/80⁺ cells) or single, live lymphocytes (for CD3⁺ and B220⁺ cells). ^∗Statistically different from CTRL, ; ^†statistically different from CTRL + HBO₂, ; ^#statistically different from DSS, . 7-AAD, 7-aminoactinomycin D.

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