Anti‐Interleukin‐16‐Neutralizing Antibody Attenuates Cardiac Inflammation and Protects against Cardiac Injury in Doxorubicin‐Treated Mice

Figures

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  Figure 1. Effects of DOX on cardiac IL-16 expression. (a, b) Cardiac IL-16 expression and serum IL-16 levels were measured in the control and DOX groups (nonparametric test). (c) Cardiac IL-16 expression in the 2 groups was determined by immunofluorescence staining (200x). (d) Effects of DOX on IL-16 mRNA expression in CTLL-2 T lymphocytes (Lyms), RAW264.7 macrophages (Møs), DC2.4 dendritic cells (DCs), and HL-1 cardiomyocytes (CMs) (Student's t-test). (e) Double immunofluorescence staining with anti-F4/80 and anti-IL-16 in DOX-induced mice (200x). N = 5 in each group. ∗p < 0.05 vs. the control group.

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  Figure 2. Effects of anti-IL-16 nAb on DOX-induced cardiac injury. (a) BW changes at different time points in the four groups (ANOVA). (b, c) HW and HW/TL ratios in each group (ANOVA). (d) HE staining and cardiomyocyte vacuolization analysis of each group (200x, nonparametric test). (e-g) Serum LDH, CK-MB, and cTnT levels were detected (ANOVA). N = 5-10 for each group. ∗p < 0.05 vs. the IgG saline group. #p < 0.05 vs. the IgG DOX group.

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  Figure 3. Effects of anti-IL-16 nAb on DOX-induced M1 macrophage differentiation. (a) Protein expression of p-p65 in both the cytoplasm and nucleus was detected in the four groups (nonparametric test). (b) Cardiac expression of CD80 (brown granules) and iNOS (brown granules) was detected by immunohistochemical staining (200x, ANOVA). (c) Cardiac mRNA levels of TNF-α, IL-1β, IL-6, IL-17, IL-18, IFN-γ, and MCP-1 were detected in each group by RT-qPCR (ANOVA). N = 5 for each group. ∗p < 0.05 vs. the IgG saline group. #p < 0.05 vs. the IgG DOX group.

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  Figure 4. Effects of anti-IL-16 nAb on DOX-induced cardiomyocyte apoptosis. (a) Protein expression of Bax, Bcl-2, cleaved caspase-3, and caspase-3 were measured in the left ventricle, and the Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were analyzed (ANOVA). (b) Cardiac TUNEL-positive cells were measured and analyzed. N = 5 for each group (200x, nonparametric test). ∗p < 0.05 vs. the IgG saline group. #p < 0.05 vs. the IgG DOX group.

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  Figure 5. Effects of anti-IL-16 nAb on DOX-induced M1 macrophage differentiation and cardiomyocyte apoptosis in vitro. (a) Nuclear p-p65 expression in RAW264.7 macrophages was detected (ANOVA). (b) Double immunofluorescence staining of anti-F4/80 and anti-IL-16 in DOX-treated mice (200x, nonparametric test). (c) The mRNA expression of CD80 and iNOS was detected (ANOVA). (d) Protein expression of Bax, Bcl-2, cleaved caspase-3, and caspase-3 was measured and the ratios of Bax/Bcl-2 and cleaved caspase-3/caspase-3 (ANOVA). N = 5 for each group. ∗p < 0.05 vs. the IgG saline group. #p < 0.05 vs. the IgG DOX group.