Maintenance of an inflammatory state by Ad36 infection despite exercise.

(A) Mice were infected with Ad36 or Ad2, or given sham injections. The mRNA levels of inflammatory cytokines MCP-1 (p<0.05, ANOVA in nonexercise; p<0.001, ANOVA in exercise) and TNF-α (p<0.05, ANOVA) in the epidermal fat pads were assessed and the levels in exercise and nonexercise mice were compared with those in Ad36-infected mice. (B) The protein samples were obtained from the epidermal fat pads. The MCP-1 (p<0.05, ANOVA) and TNF-α (p<0.05, ANOVA in nonexercise; p<0.001, ANOVA in exercise) protein levels were assayed by ELISA. (C) Stromal vascular fractions were isolated from the epidermal fat pads and stained with F4/80 antibody as a macrophage marker. The relative reduction in macrophages was calculated based on the difference between nonexercise and exercise mice (p<0.05; ANOVA). (D) The epidermal fat pads were stained with hematoxylin and eosin (arrows indicate infiltrated immune cells). (E) The mRNA levels of macrophage markers were assessed (CD64 for M1 macrophages (p<0.05, ANOVA), CD206 for M2 macrophages (p<0.05, ANOVA), A.U., arbitrary units) to compare the levels in exercise and nonexercise Ad36-infected mice with those in sham-infected mice. The statistical analysis was carried out independently for each group, with lowercase letters indicating the nonexercising group, and capital letters for the exercising group. Statistically, groups with different letters over the bars were significantly different.