FIGURE 5.

AAT protects islets cells from activated macrophage-induced apoptosis. (A) Human islet cell death after co-culture with IFN-γ-activated macrophages by TUNEL staining. Nuclei were stained with DAPI (blue), β-cells with the anti-insulin antibody (red), TUNEL⁺ cells are green. (B) Quantification of TUNEL+ cells within an islet; 15–20 islets were included in each group. (C) Islet cell death measured by the apoptosis ELISA kit. CTR, human islets; IFN, human islets co-cultured with IFN-γ-treated macrophages; IFN+AAT, human islets co-cultured with IFN-γ-treated macrophages in the presence of AAT. (D) Death of vehicle or cytokine-treated islets measured by cell apoptosis ELISA kit. (E) M1 marker gene expression in Raw264.7 cells measured at 24 hours after co-cultured with cytokine-treated islets. Vehicle, nontreated Raw264.7 cells; Cyto-Islet, cytokine (human IL-1β, TNF-α, and IFN-γ)-treated human islets; Cyto-Islet +AAT, cytokine-treated human islets in the presence of AAT. Data shown are mean ± SEM; *P < .05, CTR vs. Cyto-islets or Cyto-islets+AAT, and ^#P < .05 Cyto-islets vs. Cyto-islets+AAT, analyzed by one-way ANOVA