Ultrasound-Targeted β-Catenin Gene Therapy Improves the Cardiac Function in Mice After Myocardial Infarction

Figures

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  Fig. 1. Optimal conditions for UTMD gene transfection in vivo. A Left representative image of bioluminescence in the heart region of experimental animals after UTMD transfection of luciferase in vivo. Right representative images of fluorescence 5 days after transfection of the luciferase gene in the heart, lungs, liver, and spleen region of experimental animals. B Bioluminescence after UTMD gene transfection. Noting that peak bioluminescence observed on day 2 after transfection. ***P < 0.001. n = 4 per group. C Bioluminescence of UTMD gene transfection. Noting that the optimal frequency of UTMD gene transfection at 3 times. *P < 0.05, ***P < 0.001, vs. 1 day; ##P < 0.01, ###P < 0.001, vs. 2 days; &P < 0.05, &&&P < 0.05, vs. 3 days. n = 3 per group. UTMD, ultrasound-targeted microbubble destruction

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  Fig. 2. Effect of pre-treatment with non-invasive UTMD β-Catenin gene-targeted transfection cardiac function in mice after myocardial infarction. A Representative traces of PCR of β-catenin mRNA expression after UTMD β-catenin gene-targeted transfection in mouse heart; B Representative traces of western blots of β-catenin protein after UTMD β-catenin gene-targeted transfection in mouse heart. Lower panels pooled data for expression of β-catenin mRNA (left) and β-catenin protein (right) *P < 0.05, n = 5 per group. C Typical images of echocardiography in mice before surgery, 14 days and 28 days after surgery in the control and β-catenin gene-treated groups; (D) The measurements of left ventricular EF and FS at short axis view before surgery, 14 and 28 days after MI in the control and β-catenin-treated groups. **P < 0.01; Control group, n = 9, β-catenin group, n = 11. PCR, polymerase chain reaction; UTMD, ultrasound-targeted microbubble destruction; EF, ejection fraction; FS, fractional shortening; MI, myocardial infarction

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  Fig. 3. Effect of pre-treatment with non-invasive UTMD β-Catenin gene-targeted transfection on cardiac morphology in mice after myocardial infarction. A Left representative images of Masson’s trichrome staining regions of infarct in the heart 14 days after MI in the control and β-catenin gene pre-treated groups. Scale bar = 50 mm; right representative images of hearts with infarct in the heart 28 days after MI in the control and β-catenin gene pre-treated groups; Lower panels representative images of regions of infarct in the heart 14 and 28 days after MI in the control and β-catenin gene pre-treated groups. B Area and wall thickness of infarct zones in hearts 14 and 28 days after MI in the control and β-catenin gene pre-treated groups. *P < 0.05; **P < 0.01. n = 6 per group. UTMD, ultrasound-targeted microbubble destruction; MI, myocardial infarction

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  Fig. 4. Effect of UTMD β-Catenin gene-targeted transfection on cardiac function and the number of cardiac stem cells in mice after myocardial infarction. A TUNEL was used to determine myocardial tissue apoptosis. B–C Left immunofluorescent images of new myocardial arterioles (B) and capillaries (C) of the infarct border zone of the UTMD β-catenin gene pre-treated and control mice 28 days after MI. Red fluorescence indicating labeled with anti-α-SMA/anti-factor VIII antibodies, and blue fluorescence representing nuclei stained with DAPI, Scale bar = 50 μm; right pooled data for numbers of new myocardial arterioles of the UTMD β-catenin gene pre-treated and control mice 28 days after MI. **P < 0.01, n = 9 per group. D Flow cytometry analysis of C-kit + /CD34 + and Scal + /CD34 + SCSs of the control and β-catenin gene-treated groups; Flow cytometry analysis of C-kit + /CD34- and Scal + /CD34- SCSs of the control and β-catenin gene-treated groups. *P < 0.05; **P < 0.01, n = 4 per group. E Upper representative images of colonies of CFU and CFU-F cultured by cardiac single cells from the control and β-catenin gene-treated groups. Scale bar = 50 μm; below pooled data of colonies of CFU and…