Ozone Exerts Cytoprotective and Anti-Inflammatory Effects in Cardiomyocytes and Skin Fibroblasts after Incubation with Doxorubicin

Figures

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  Figure 1. Cardiomyocyte (a) and skin fibroblast (b) cell viability (±SEM) performed by the modified MTT method after incubation for 24 h with doxorubicin and ozone-doxorubicin coexposure. ∗p < 0.001; ∗∗p < 0.05; ns: not significant.

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  Figure 2. The MMP of cardiomyocytes (a) and skin fibroblasts (b) determined by a JC-1 kit, relative to the control (untreated) cells. Values are presented as the mean ± standard error of the mean (n = 3).

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  Figure 3. Effects of ozone on NF-κB activation. Human fetal cardiomyocytes (a) and skin fibroblasts (b) were pretreated with various concentrations of ozone (10, 20, 30, 40, and 50 μg/mL) and doxorubicin for 24 h. Nuclear extracts were prepared, and NF-κB activation was measured by enzyme-linked immunosorbent assay. Quantified data are expressed as the mean ± standard error of the mean of three experiments. ∗p < 0.001; ∗∗p < 0.05; ns: not significant.

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  Figure 4. Leukotriene B4 expression (pg/mL) in cardiomyocytes (a) and skin fibroblasts (b) exposed to arachidonic acid (10 μM), doxorubicin (100 nM) alone, or coincubated with ozone at 10, 20, 30, 40, and (50 μg/mL). ∗p < 0.001; ∗∗p < 0.05; ns: not significant.

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  Figure 5. Nrf2 binding activity in cardiomyocytes and skin fibroblasts by doxorubicin (100 nM) and ozone-doxorubicin treatments. Values are expressed as fold change in Nrf2 activity from untreated (control) cells.

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  Figure 6. Anti-inflammatory properties of ozone at different concentrations on IL-1, IL-8, IL-6, TNF-α, MMP-2, and MMP-9 production by cardiomyocytes and skin fibroblasts (at a density of 1.2 × 105 cells/well). Cells were treated with or without ozone (from 10 up to 50 μg/mL) for 5 h before exposure to doxorubicin at subclinical concentration (100 nM) for 12 h ∗p < 0.001; ∗∗p < 0.05; ns: not significant.