Angiopoietin-1 Promotes the Integrity of Neovascularization in the Subcutaneous Matrigel of Type 1 Diabetic Rats

Figures

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  Figure 1. Effect of Ang-1 on neovascularization in diabetic rats. (a) Observe the appearance of the Matrigel. (b) H&E staining of Matrigel. Black arrows represent inflammatory cells, ∗ represents neovascularization, ×200. (c) STZ-induced type 1 diabetes in rats with blood glucose and urinary protein, where △ represents P <0.05 compared with the control group.

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  Figure 2. Electron micrographs of the newly formed microvessels in the Matrigel. Ultrastructural characterization of microvessels in the diabetes group of Matrigel; the microvessels have poorly organized basement membrane and swollen endothelial cells, as well as damaged endothelial tissue (Black arrow). Few pericytes are distributed around the endothelium, magnification ×20000. Ang-1 treated Matrigel showed complete endothelial structure with the typical tight junction (black arrows). Pericytes were observed to be in direct contact with endothelial cells. Multiple elongated pericytes are embedded within the intact basement membrane, magnification ×20000.

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  Figure 3. Immunofluorescence analysis of pericyte coverage of microvessels in the Matrigel. (a) Ang-1 treatment of diabetic rats; the neovascular marker JG12 (red) and the expression of desmin (green), a pericyte marker, were confirmed by immunofluorescence, 50μm. (b) New angiogenic marker JG12 (red) and pericyte α-SMA (green) expression were confirmed by double-labeling technique, 50μm.

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  Figure 4. Vascular maturity-related protein expression after Ang-1 treatment. (a) Immunohistochemical staining of ZO-1, occludin, VE-cadherin, and Connexin40 in the four groups was performed. Several proteins are expressed on the envelope of endothelial cells (black arrow), × 200. (b) The relative quantification of ZO-1, occludin, VE-cadherin, and Connexin40 in four groups. △ represents P <0.05 compared with the control group, ∗ represents P <0.05 compared with the Ang-1 treatment group, and N/D represents no expression.

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  Figure 5. Analysis of Vascular maturity-related protein expression by RT-PCR. The mRNA expression of α-SMA, desmin, ZO-1, VE-cadherin, and Connexin40 was detected. △ represents P <0.05 compared with the control group, ∗ represents P <0.05 compared with the Ang-1 treatment group, and N/D represents no expression.