Intraperitoneal Injection of Human Ferritin Heavy Chain Attenuates the Atherosclerotic Process in APOE-Knockout Mice

Figures

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  Figure 2. Analysis of aortic plaque and aortic revascularization in 2 groups of mice. (A) Magnetic resonance imaging and reconstruction results of the abdominal aortas of mice at 20 weeks. (B) Differential analysis of relevant parameters of aortic structure in 2 groups of mice. (C) Results of oil-red O staining and analysis of aorta in 2 groups of mice. ** p < 0.01, *** p < 0.001, **** p < 0.0001, black dots, visualization of numerical results.

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  Figure 1. Illustration of the experimental procedure and analysis of HE staining and Masson staining of aortic plaques in Apo-E(−/−) mice. (A) Two groups of mice were fed on a high-fat diet for 20 weeks. FT-H was administered intraperitoneally to the injection group at week 10 for 10 weeks. At week 20, mice in both groups underwent abdominal aortic MRI, and tissue and blood sampling was performed at the end of imaging. (B) HE staining and Masson staining (10× magnified field of view and 40× magnified field of view) of the aortas of both groups of mice at 20 weeks. Collagen is blue in color and is mainly deposited between the intima and mesentery of the artery. Bar = 100 μm. (C) Graph showing the CVF value analysis results from the Masson staining of aortas in the 2 groups of 8 mice. * p < 0.05, black dots, visualization of numerical results. (D) Images show the HE staining results of the heart, liver, spleen, lungs and kidneys of both groups of mice.

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  Figure 3. Serum biochemical and metabolic parameters of 8 mice in the 2 groups at 20 weeks. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, black dots, visualization of numerical results.

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  Figure 4. Expression analysis of H-FT, L-FT and TfR1 in the aortas of mice from 2 groups. (A) Immunohistochemical results show that the iron metabolism proteins H-FT, L-FT and TfR1 were mainly distributed in the carotid intima and carotid mesentery (n = 4). Positive staining was brownish yellow. Bar = 100 μm. (B) Graph showing the results of the immunohistochemical AOD value analysis of H-FT, L-FT and TfR1 in the aortas of 8 mice from the 2 groups. * p < 0.05, ** p < 0.01, black dots, visualization of numerical results.

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  Figure 5. Prussian blue staining of aorta in 2 groups of mice, and analysis of NRF-2 and GPX-4 expression. (A) Prussian blue staining showed that the area of iron-positive region in the aortas of mice in the injection group was larger than that in the control group. (B–E) Immunohistochemical results showing the distribution of NRF-2 and GPX-4 and the results of AOD values in both groups of mice. Positive staining was brownish yellow. Bars = 100 μm. * p < 0.05, black dots, visualization of numerical results.

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  Figure 6. Expression analysis of MMP-2, MMP-9 and MCP-1 in the aortas of mice from the 2 groups. (A) Immunohistochemical results show that the iron metabolism proteins MMP-2, MMP-9 and MCP-1 were mainly distributed in the carotid intima and carotid mesentery (n = 4). Positive staining was brownish yellow. Bar = 100 μm. (B) Graph showing the results of the immunohistochemical AOD analysis of MMP-2, MMP-9 and MCP-1 in the aortas of a total of 8 mice from the 2 groups. * p < 0.05, black dots, visualization of numerical results.