Figure 6.

Protective role of PEP–TPP–mitochondria in myocardial IR injury by inhibiting apoptosis, immune cell infiltration, and proinflammatory reaction. (A) Cleaved caspase 3 levels and the ratios of pro-apoptotic Bax and anti-apoptotic Bcl-2 were analyzed by Western blotting in mice heart. Sham (left), IR (middle), and IR and PEP–TPP–mitochondria transplantation (IR+PEP-MITO, right). (B) Statistical assay of relative expression levels of cleaved caspase 3. (C) Statistical assay of ratios of pro-apoptotic Bax and anti-apoptotic Bcl2. (D,E) Apoptotic cardiomyocytes (red) were quantified by TUNEL assay after PEP–TPP–mitochondrial compound transplantation and myocardial IR injury. Cell nuclei were stained by DAPI (blue). Scale bars, 50 μm (n ≥ 3 per group). (F) Flow cytometry analysis of single-cell suspension isolated from fresh heart tissues after staining for macrophage markers CD45, F4/80, and CD11b. (G) Statistical significance of CD45+CD11b+F4/80+ macrophages. (H–K) Western blots and statistical analyses for expression changes of proinflammatory cytokines, including NLRP3, IL6, and cleaved IL 1β. Mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001. Statistical analysis was carried out by a one-way ANOVA analysis followed by Tukey’s test for post hoc analysis.