MitoQ Ameliorates Diabetic Cardiomyopathy by Inhibiting the mtROS–TXNIP–NLRP3 Pathway

Figures

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  Figure 1. MitoQ improves the degree of cardiac fibrosis and inhibits the decline of cardiac function caused by diabetes. (A) TUNEL staining of mouse heart tissue (20×). (B) Quantitative analysis of TUNEL staining results of cardiac tissue. (C) HE staining and Masson staining of mouse heart tissue (20×). (D) Quantification of myocardial fibrosis based on Masson’s staining. (E) Representative echocardiographic images of mouse hearts. (F) Left ventricular fractional shortening (LVFS) in each group. (G) Left ventricular ejection fraction (LVEF) in each group. Data are presented as mean ± SEM. For animal experiments, n = 8 mice per group. ∗∗ p < 0.05.

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  Figure 2. MitoQ alleviates oxidative stress and inhibits activation of the TXNIP/NLRP3 pathway in diabetic mouse hearts. (A) Detection of total SOD content in mouse myocardial tissue. (B) Detection of total antioxidant capacity in mouse myocardial tissue. (C) Immunohistochemical detection of TRX, TXNIP, NLRP3, Caspase‐1, and cleaved IL‐1β protein expression in mouse heart tissue (20×). (D) Quantitative analysis of immunohistochemical staining results. Data are presented as mean ± SEM. For animal experiments, n = 8 mice per group. ∗∗ p < 0.05.

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  Figure 3. MitoQ reduces mitochondrial ROS production in AC16 cardiomyocytes under high‐glucose conditions. (A) CCK‐8 assay showing cell viability of AC16 cardiomyocytes treated with different concentrations of MitoQ under 5.5 mM (normal glucose) and 30 mM (high glucose) conditions. (B) Time‐dependent effects of different concentrations of MitoQ on AC16 cell viability under high‐glucose conditions, assessed by CCK‐8 assay. (C) Representative images of mitochondrial membrane potential in AC16 cells (20×). (D) Quantification of mitochondrial membrane potential fluorescence intensity. (E) Representative images of mitochondrial ROS production in AC16 cells (20×). (F) Quantitative analysis of mitochondrial ROS fluorescence intensity. Data are presented as mean ± SEM. CCK‐8 assays were performed with n = 9 measurements per group. ∗∗ p < 0.05.

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  Figure 4. MitoQ inhibits the activation of NLRP3 inflammasome in AC16 cells under high‐glucose conditions. (A) Representative co‐immunoprecipitation (Co‐IP) and western blot analyses showing the interaction among TRX, TXNIP, and NLRP3 proteins in AC16 cells. (B) Quantitative analysis of TRX, TXNIP, and NLRP3 protein expression levels in the input group. (C) Quantitative analysis of TRX–TXNIP and TXNIP–NLRP3 interactions in the IP group. (D) Representative immunofluorescence images of NT ‐GSDMD, Caspase‐1, and cleaved IL‐1β expression in AC16 cells. (E) Quantitative analysis of immunofluorescence intensity. (F) Total SOD content in AC16 cells. (G) Total antioxidant capacity in AC16 cells. Data are presented as mean ± SEM; WB and Co‐IP, n = 3 independent experiments; SOD and total antioxidant capacity assays, n = 9 measurements per group. ∗∗ p < 0.05.

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  Figure 5. Combined TXNIP knockdown and MitoQ treatment attenuates high glucose‐induced NLRP3 inflammasome activation and cardiomyocyte death. (A) Representative co‐immunoprecipitation (Co‐IP) analysis showing the interaction among TRX, TXNIP, and NLRP3 proteins in AC16 cells. (B) Quantitative analysis of TRX –TXNIP and TXNIP–NLRP3 interactions in the IP group. (C) Representative western blot analysis of NT‐GSDMD, Caspase‐1, and cleaved IL‐1β expression in AC16 cells. (D) Quantitative analysis of western blot results. (E) Representative propidium iodide (PI) staining images showing cardiomyocyte death. (F) Quantitative analysis of PI test results. Data are presented as mean ± SEM; WB and Co‐IP, n = 3 independent experiments. ∗∗ p < 0.05.