Expression of integrin-linked kinase improves cardiac function in a swine model of myocardial infarction

Figures

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  Figure 1.. Establishing a swine myocardial infarction model. (A) Pigs underwent 90 min proximal left anterior descending coronary artery balloon occlusion. (B) Preoperative ECG. (C) ST elevation in operation. (D) ECG of post-operation. (E) Heart slices of sham-operation. (F) Heart slices of infarct area with white scar. (G) Serum cTnI levels of pre- and post-surgery, *compared with before operation, P<0.01. (H and I) Heart section staining by H&E indicated cardiomyocyte necrosis. Scale bars, 100 µm.

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  Figure 2.. Expression of ILK and hrGFP four weeks after adenoviral delivery. (A) Immunofluorescence for α-actin and DAPI in cardiomyocytes. (B) Immunofluorescence for hrGFP expression in the myocardium. (C) hrGFP expression appeared in α-actin-positive cells, indicating successful Ad-ILK transfection into cardiomyocyte. (D) Western blotting showing hrGFP expression in the infarction zone four weeks after adenoviral delivery compared with null control. GAPDH expression is shown as internal control. (E and G) Western blotting showing ILK expression after adenoviral delivery in the infarction zone compared with null control. (F and H) Western blotting showing ILK expression after adenoviral delivery in the infarction remote zone compared with null control. GAPDH expression is shown as internal control. *P>0.05, n=6. Scale bars, 40 µm. ILK, integrin-linked kinase; hrGFP, humanized recombinant green fluorescent protein; DAPI, 4′,6-diamidino-2-phenylindole.

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  Figure 3.. Heart imaging studies in mini pig MI. (A-D) LVEDD, IVSD, EF and LVPWd improved in the ad-ILK group compared with the ad-null group. n=6, *P<0.05. (E and F) Ad-ILK improves heart function four weeks post-MI. The myocardial perfusion image by ECT and the myocardial metabolic image by PET. LVEDD, LV end-diastolic diameter; IVSD, interventricular septal thickness in diastole; MI, myocardial infarction; PET, positron emission tomography.

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  Figure 4.. Overexpression of ILK prevents apoptosis and increases microvessel density in the infarct zone. (A and B) Triple-staining with TUNEL (green), anti-α-sarcomeric actin antibody (red) and DAPI (blue) in the infarction zone of (A) ad-ILK and (B) ad-null. Scale bars, 50 µm. (C and D) Microvessel density is measured by frozen sections stained with vWF in the infarct zone of (C) ad-ILK and (D) ad-null group; scale bars, 25 µm. (E) Quantification of TUNEL-positive cardiomyocytes in the infarct zones of ad-ILK and ad-null animals. n=6 per group. *P<0.01. (F) Quantification of microvessel density in faction zone of ad-ILK and ad-null animals. n=6 per group. *P<0.01.