Ultrasonic Microbubble Cavitation Deliver Gal-3 shRNA to Inhibit Myocardial Fibrosis after Myocardial Infarction

Figures

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  Figure 1. Schematic representation of Gal-3 shRNA delivery mediated cardiac function protection. The arrow represents downregulation (This figure was drawn by Figdraw).

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  Figure 2. Characterization of Gal-3 shRNA/CMBs. (A) Schematic representation of Gal-3 shRNA/CMBs preparation. (B) Representative optical micrographs of CMBs and Gal-3 shRNA/CMBs. (C) Appearance of the CMBs and Gal-3 shRNA/CMBs. (D) Size distribution of CMBs and Gal-3 shRNA/CMBs. (E) Average particle size of CMBs and Gal-3 shRNA/CMBs. (F) Average Zeta potentials of CMBs and Gal-3 shRNA/CMBs. (n = 3, ** p < 0.01 *** p < 0.001).

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  Figure 3. Gal-3 expression in myocardium. (A) Therapy regimen schematic of Ultrasonic Microbubble Cavitation-mediated Gal-3 shRNA delivery in MI rats. (B) HE staining and Masson trichrome staining of hearts sectioned at mid-ventricle at 0, 1, 3, 7, 14, 21, 28 days post-MI. (C) Temporal changes in Gal-3 mRNA expression post-MI. (D) Myocardial Gal-3 mRNA level in Gal-3 shRNA/CMBs + US group and control group. (E) Gal-3 protein expression in Gal-3 shRNA/CMBs + US group and control group. (F) Immunohistochemistry staining with Gal-3+ cells on day 21. (G) Concentration of Gal-3 in serum. Infarcted represents the infarcted area, border represents the border zone, and non-infarcted represents the non-infarcted area (n = 3, ns p > 0.05, * p < 0.05, ** p < 0.01).

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  Figure 4. The improvement in cardiac function in MI rats. (A) Representative M-mode echocardiography images. The symbol showed the measurement of cardiac function. (B,C) Cardiac function indicators of LVEF, FS, LVIDd and LVIDs were evaluated accordingly. (n = 3, ** p < 0.01).

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  Figure 5. Myocardial fibrosis and macrophage infiltration in MI rats. The blue-stained area represents the fibrous tissue, and the red-stained area represents the myocardial tissue. (A) Masson staining and quantitative analysis of scar area in the two groups on day 7 and day 21. (B) Degree of fibrosis and quantitative analysis of the fibrosis ratio in three regions between the two groups on day 21 (C) Immunohistochemistry staining with CD68+ cells on day 21 (n = 3, ns p > 0.05, * p < 0.05, ** p < 0.01).

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  Figure 6. Collagen synthesis in MI rats. (A) COL1A1 mRNA expression in different regions of MI heart. (B) COL3A1 mRNA expression in different regions of MI heart. (C) Ratio of Col1/Col3 mRNA expression in different regions of MI heart (n = 3, * p < 0.05, ** p < 0.01).