Fig. 6: Inflammation levels in brain tissues after clusterzyme treatment.

a–c Western blotting for IL-1β, IL-6, and TNFα in the brain tissues 1, 3, and 7 days post TBI after treatment (n = 3 per group), respectively. d Western blotting quantitative analysis of inflammatory factors at different time points (n = 3 per group). All the samples were derived from the same experiment and blots were processed in parallel. Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). It can be seen that Au₂₄Cd₁ can rapidly and significantly reduce the upregulated inflammatory cytokines of IL-1β and IL-6 after brain injury, while Au₂₄Cu₁ has a better ability to reduce the expression of TNFα. e–g ELISA quantitative analysis of IL-1β, IL-6, and TNFα levels in brain tissues on days 1, 3, and 7 with or without clusterzymes treatment (n = 5 per group), respectively. Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). h Immunofluorescence co-staining of IL-1β and microglia (Iba-1), astrocytes (GFAP), or neurons (NeuN) in injured cortex 3 days post injury with or without clusterzyme treatment. Quantitative analysis of i the number of IL-1β⁺ expression in different positive cells and j the pixels density of Iba-1/NeuN/GFAP cells in the injured cortex with or without clusterzyme treatment (n = 3 per group). Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). Experiments were repeated independently a–c twice and h three times with similar results.