A tripartite therapeutic agent reprograms the myocardial infarction microenvironment

Figures

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  Scheme 1. Schematic illustration of MMRA treatment for MI via anti-inflammatory, antioxidative, and conductive properties for myocardial repair.

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  Fig. 1. Synthesis and characterization of MMRA. (A) SEM image of PPy. (B) FTIR spectra of PPy. (C) UV-Vis-NIR spectra of Sino, PPy and MMRA. (D) SEM images of MMRA under different magnifications. (E) Rheological behavior of P407 hydrogel and MMRA under elevated temperature conditions. (F–G) Rheological analysis of the hydrogels at 37 °C in time sweep mode and amplitude sweep mode. (H) Gelation time of P407 and MMRA (n = 3). (I) Conductivity of P407 and MMRA (n = 3). (J) Photographs of an LED circuit with P407 and MMRA. (K)The sol−gel−sol transition of P407 and MMRA at various temperatures; Photograph of the MMRA injected into DI water through a needle. (L–N) SOD activity, ABTS and DPPH scavenging activities of Sino, PPy, Sino-PPy solution and MMRA. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 2. The protective effects of MMRA on H2O2-induced oxidative stress and LPS-induced inflammation. (A–C) The level of MDA, GSH and SOD in H9C2 cells exposed to H2O2 (n = 3). (D) The ROS levels were determined by DHE (red) staining in H9C2 cells (n = 3). (E) Live/dead staining in H9C2 cells (n = 4). (F, G) Representative immunofluorescence staining of iNOS and CD206 in the LPS-induced inflammation in RAW264.7 cells (n = 3). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 3. MMRA treatment improves cardiac function in MI mice. (A) Representative images of cardiac echocardiography. (B–E) Quantitative analysis of LVEF%, LV-FS%, LVIDs and LVIDd (n = 7). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 4. MMRA treatment reduces infarct size. (A)TTC staining schematic diagram of tissue sections. (B–C) Representative tissue sections and quantitative analysis of infarct area (n = 5). (D) TUNEL staining of myocardial tissue (n = 6). ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 5. Antioxidant and electrical integration effects of MMRA after MI. (A) Representative DHE staining images and quantitative analysis of ROS level (n = 6). (B) Representative images of mitochondrial morphology using TEM. (C–D) Representative fluorescence images and quantitative analysis of CX43 and α-actinin (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 6. MMRA regulates cardiomyocyte inflammation in MI. (A) Representative images of H&E staining. (B–C) Representative immunofluorescence images and quantification of CD68 and iNOS (n = 6). (D–G) qPCR analysis of the levels of cytokines, including TNF-α, IL-1β, TGF-β and IL-10 (n = 6). (H) Representative immunohistochemical staining and quantification of CD206 (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 7. MMRA inhibits myocardial fibrosis After MI. (A) Representative Masson's trichrome staining images. (B) Representative images of Sirius Red staining. (C) Masson's trichrome staining quantitative analysis (n = 6). (D–E) Representative WGA immunofluorescence staining images and quantitative analysis of cardiomyocyte cross-sectional area (n = 6). (F) Representative immunofluorescence staining images of Collagen I and III and quantitative analysis (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

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  Fig. 8. Transcriptomic and Bioinformatics Analysis of MMRA in MI. (A) Schematic diagram of the transcriptomic animal treatment protocol. (B) DEGs statistics of between the MI and the MI + MMRA group. (n = 5). (C) Heatmap. (n = 5). (D–E) GO annotations and enrichment analysis. (n = 5). (E–F) KEGG annotations and enrichment analysis. (n = 5).