Elastic fibers reconstructed using adenovirus-mediated expression of tropoelastin and tested in the elastase model of abdominal aortic aneurysm in rats

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Presentation transcript:

Elastic fibers reconstructed using adenovirus-mediated expression of tropoelastin and tested in the elastase model of abdominal aortic aneurysm in rats Jiang Xiong, MD, Shen Ming Wang, MD, Liu Hua Chen, MD, Ying Lin, MD, Yi Fan Zhu, MD, Cai Sheng Ye, MD Journal of Vascular Surgery Volume 48, Issue 4, Pages 965-973 (October 2008) DOI: 10.1016/j.jvs.2008.04.016 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 The morphology and histology of the aneurysmic rat aorta with and without adenoviral tropoelastin green fluorescent protein (AdTE-GFP) treatment (A) aortic diameters (AD) measurements for rats perfused with elastase and treated with AdTE-GFP, empty adenovirus (AdNull), or phosphate buffered saline (PBS) (N equals 4/each experiment). (B) Frozen section of aneurysmal wall at day 3 after transfection with AdTE-GFP, visualized by fluorescence microscopy (original magnification ×200). (C) The same section shown in B but stained with HE stain (original magnification ×200). (D-I) Histological analysis of the aortic wall of rats after elastase perfusion and transfection with AdTE-GFP (D-G), or AdNull-GFP (H, I). The von Giesson stain for elastin is shown on the left and the GFP antibody stain is shown on the right. The VSMCs transfected with AdTE-GFP stained immunopositive (brown) with GFP antibody. The red arrowheads indicate the VSMCs in the medial layer transfected with AdTE-GFP and stained brown with GFP antibody. (For the inset in D, F, and H original magnification ×40, original magnification for D, F, and H × 100, original magnification for E, G, and I × 200). Journal of Vascular Surgery 2008 48, 965-973DOI: (10.1016/j.jvs.2008.04.016) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 The morphology and histology of the aneurysmic rat aorta with and without adenoviral tropoelastin green fluorescent protein (AdTE-GFP) treatment (A) aortic diameters (AD) measurements for rats perfused with elastase and treated with AdTE-GFP, empty adenovirus (AdNull), or phosphate buffered saline (PBS) (N equals 4/each experiment). (B) Frozen section of aneurysmal wall at day 3 after transfection with AdTE-GFP, visualized by fluorescence microscopy (original magnification ×200). (C) The same section shown in B but stained with HE stain (original magnification ×200). (D-I) Histological analysis of the aortic wall of rats after elastase perfusion and transfection with AdTE-GFP (D-G), or AdNull-GFP (H, I). The von Giesson stain for elastin is shown on the left and the GFP antibody stain is shown on the right. The VSMCs transfected with AdTE-GFP stained immunopositive (brown) with GFP antibody. The red arrowheads indicate the VSMCs in the medial layer transfected with AdTE-GFP and stained brown with GFP antibody. (For the inset in D, F, and H original magnification ×40, original magnification for D, F, and H × 100, original magnification for E, G, and I × 200). Journal of Vascular Surgery 2008 48, 965-973DOI: (10.1016/j.jvs.2008.04.016) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 The expression of tropoelastin (TE) has obviously increased after adenoviral tropoelastin green fluorescent protein (AdTE-GFP) treated: Real-time Quantitative polymerase chain reaction (PCR) analyses were carried out for the transcriptional level of TE from the vascular smooth muscle cells (VSMCs) transfected with AdTE-GFP, empty adenovirus (AdNull), or phosphate buffered saline (PBS) at 1 day, 3 days, and 5 days (A), and in rats treated with the same constructs at 2 weeks and 4 weeks (B). All measured data were normalized to β-actin by calculating a relative expression level defined as the ratio of expression of the target gene over the expression of the housekeeping gene (β-actin) in a given sample. N equals 3/experiment (VSMCs' experiment) and N equals 4/experiment (Rat's experiment). **P < .01; *P < .05; #P > .05. Journal of Vascular Surgery 2008 48, 965-973DOI: (10.1016/j.jvs.2008.04.016) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 The tropoelastin (TE) protein is expressed at vascular smooth muscle cells (VSMCs) or rats after adenoviral tropoelastin green fluorescent protein (AdTE-GFP) treated. Immunoblot analysis of TE protein from the VSMCs transfected with AdTE-GFP, empty adenovirus (AdNull) or phosphate buffered saline (PBS) at 1 day, 3 days, and 5 days (A), and from rats treated with the same constructs at 2 weeks and 4 weeks (B). Euthanasia (E), AdTE-GFP; N, AdNull; (C) Control PBS. (C and D) showed the relative expression level of TE protein that defined as the ratio of integrated optical density (OD) of TE protein band over the integrated OD of β-actin band from the corresponding samples in (A and B), respectively. (E, F, G) The VSMCs expressing GFP were visualized by fluorescence microscope at day 1, 3, and 5 (original magnification ×400) after AdTE-GFP virus transfection. **P < .01; *P < .05. Journal of Vascular Surgery 2008 48, 965-973DOI: (10.1016/j.jvs.2008.04.016) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 The tropoelastin (TE) protein is expressed at vascular smooth muscle cells (VSMCs) or rats after adenoviral tropoelastin green fluorescent protein (AdTE-GFP) treated. Immunoblot analysis of TE protein from the VSMCs transfected with AdTE-GFP, empty adenovirus (AdNull) or phosphate buffered saline (PBS) at 1 day, 3 days, and 5 days (A), and from rats treated with the same constructs at 2 weeks and 4 weeks (B). Euthanasia (E), AdTE-GFP; N, AdNull; (C) Control PBS. (C and D) showed the relative expression level of TE protein that defined as the ratio of integrated optical density (OD) of TE protein band over the integrated OD of β-actin band from the corresponding samples in (A and B), respectively. (E, F, G) The VSMCs expressing GFP were visualized by fluorescence microscope at day 1, 3, and 5 (original magnification ×400) after AdTE-GFP virus transfection. **P < .01; *P < .05. Journal of Vascular Surgery 2008 48, 965-973DOI: (10.1016/j.jvs.2008.04.016) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions