Melatonin may be associated with changes in HUWE1, AIM2 and PANoptosis‐related proteins following myocardial ischaemia–reperfusion in male rats

Figures

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  FIGURE 1. Graphical summary. Symbols indicate changes in the IR group versus Sham (first symbol) and in the IR+Mel group versus IR (second symbol): +, significant increase; −, significant decrease; ns, no statistically significant difference. AIM2, absent in melanoma 2; APAF1, apoptotic peptidase‐activating factor 1; CASP1/8, caspase‐1/8; HUWE1, HECT, UBA and WWE domain‐containing E3 ubiquitin protein ligase 1; IR, ischaemia–reperfusion; MLKL, mixed lineage kinase domain‐like protein; RIPK1/3, receptor‐interacting protein kinase 1/3.

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  FIGURE 2. Effect of melatonin administration on infarct size. aSignificant difference compared to ischaemia–reperfusion, P = 0.021, n = 7. IR, ischaemia–reperfusion; Mel, melatonin (10 mg/kg).

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  FIGURE 3. The changes of the serum AIM2 levels across experimental groups. Individual data points for each animal are displayed as black diamonds within the boxes (n = 7 per group). In the box‐and‐whisker plots, the horizontal line within each box represents the median, the box boundaries indicate the 25th and 75th percentiles, and the whiskers extend to the minimum and maximum values. Statistical analyses were performed using one‐way ANOVA followed by Tukey's post hoc test for multiple comparisons. Groups sharing the same superscript letters (a–c) are not significantly different from each other, whereas groups with different letters differ significantly. AIM2, absent in melanoma 2; Sham, sham‐operated control group; Sham‐Mel, sham + melatonin treated group; Sham‐Vehicle, sham + carrier solution control group; IR, ischaemia–reperfusion group; IR+Mel, ischaemia–reperfusion + melatonin treated group.

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  FIGURE 4. The changes of the protein expression of APAF1 (a), RIPK3 (b), GSDMD (c), and HUWE1 (d), alongside a representative Western blot image (e). Individual data points for each animal are represented as black dots within the bars (n:7 per group). Densitometric data are expressed as mean ± SD and presented as a percentage of the control group. Statistical analyses were performed using one‐way ANOVA followed by Tukey's post hoc test for multiple comparisons. Groups sharing the same superscript letters (a–d) are not significantly different from each other, whereas groups with different letters differ significantly. Panel E shows a representative image of the target proteins and the β‐actin loading control, which is identical for all blots as the same biological samples were used across all panels. GSDMD, gasdermin D; Sham, sham‐operated control group; Sham‐Mel, sham + melatonin treated group; Sham‐Vehicle, sham + carrier solution control group; IR, ischaemia–reperfusion group; IR+Mel, ischaemia–reperfusion + melatonin treated group.

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  FIGURE 5. Effects of melatonin treatment on miR‐136 (a) and miR‐325 (b) expression levels. Individual data points for each animal are represented as black dots within the bars (n = 7 per group). Data are expressed as means ± SD. Statistical analyses were performed using one‐way ANOVA followed by Tukey's post hoc test for multiple comparisons. Groups sharing the same superscript letters (a–e) are not significantly different from each other, whereas groups with different letters differ significantly (P < 0.05). Sham, sham‐operated control group; Sham‐Mel, sham + melatonin treated group; Sham+Vehicle, sham + carrier solution control group; IR, ischaemia–reperfusion group; IR+Mel: ischaemia–reperfusion + melatonin treated group.