Fig. 4

Hydrogen decreased microglia activation that was initially induced by plantar incision surgery in the spinal cord of mice. A Confocal images of MMP-9 (green) and its colocalization with microglia (IBA-1, red) in the spinal cord after administering HRS induced by planter incision surgery; n = 6. The white arrows indicate IBA-1 colocalization with MMP-9. B Immunofluorescence staining analysis after HRS treatment. C Immunohistochemistry images of IBA-1 in the spinal cord of the mice and IHC score values of the mice at 5 days; n = 4. The black arrows indicate IBA-1. D Immunohistochemistry staining analysis after HRS treatment. E IBA-1 expression was measured in the spinal cord of mice (n = 6). Lumbar spines (L4–L5) were collected and analyzed 5 days after surgery. Scale bar: 200 µm. HRS = hydrogen-rich saline; PI = plantar incision; Veh = vehicle. A significant difference is revealed following one-way ANOVA (*P < 0.05, **P < 0.01, ***P < 0.001 vs. sham; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs. PI group; Bonferroni post hoc tests)