Construction and evaluation of a combined model of NAA/STZ-induced type 2 diabetes with carotid balloon injury in SD rats

Figures

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  Figure 1. Experimental grouping and protocol. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus; NAA, nicotinamide; STZ, streptozotocin.

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  Figure 2. Comparison of average food intake. (A), Water intake (B), excretion (C), body weight (D), and random blood glucose (E) in rats after modeling. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus; NAA, nicotinamide; STZ, streptozotocin. Two-way ANOVA was performed for statistical comparison, and Geisser–Greenhouse correction was adopted if the sphericity assumption was violated: *: Comparison between the T2DM group and the control group (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). & Comparison between the T2DM+VBI group and the control group (&P < 0.05; &&P < 0.01; &&&P < 0.001; &&&&P < 0.0001); ¥: Comparison between the VBI group and the control group (¥¥P < 0.01); #: Comparison between the T2DM+VBI group and the T2DM group (#P < 0.05; ##P < 0.01; ###P < 0.001; ####P < 0.0001).

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  Figure 3. Comparison of carotid artery histomorphology (HE Staining) after balloon injury. (A) Representative HE-stained images of the carotid artery at 2, 4, and 6 weeks post-balloon injury in each group. (B) Comparison of the neointimal area at different time points after surgery among the groups. (C) Comparison of the arterial proliferation index at different time points after surgery among the groups. (D) Comparison of the percentage of carotid artery stenosis at different time points after surgery among the groups. (E) Comparison of the arterial media area at different time points after surgery among the groups. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus. Scale bar = 100 μm. Two-way ANOVA was performed for statistical comparison, and Geisser–Greenhouse correction was adopted if the sphericity assumption was violated, *P < 0.05; **P < 0.01; ***P < 0.001.

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  Figure 4. Comparison of carotid artery histopathological and immunohistochemical staining at 6 weeks post-balloon injury. (A) Representative images of immunohistochemical staining of the carotid artery in each group. (B) Ratio of the α-SMA-positive area to the neointimal area. (C) Ratio of the F4/80-positive area to the neointimal area. (D) Ratio of the MMP-9-positive area to the neointimal area. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus; α-SMA, α-smooth muscle actin; F4/80, macrophage marker; MMP, matrix metalloproteinases. Scale bar = 50 μm. Unpaired t tests were used for statistical comparisons, and Welch’s correction was applied when variances were unequal, *P < 0.05, **P < 0.01.

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  Figure 5. HE-stained sections of major organs from each group at 6 weeks post-VBI. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus. Scale bar = 50 μm.

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  Figure 6. Analysis and comparison of whole blood cell and plasma biochemical parameters in diabetic rats at 6 weeks post-VBI. (A) RBC: red blood cell count analysis; (B) WBC: total white blood cell count analysis; (C) HGB: hemoglobin quantitative analysis; (D) PLT: platelet count analysis; (E) ALB: plasma albumin measurement; (F) ALT: alanine aminotransferase assay; (G) AST: aspartate aminotransferase assay; (H) SCr: plasma creatinine measurement; (I) BUN: plasma urea nitrogen determination; (J) TC: plasma total cholesterol concentration; (K) LDL-C: plasma low-density lipoprotein cholesterol measurement. VBI, vascular balloon injury; T2DM, type 2 diabetes mellitus. One-way ANOVA with Tukey’s post hoc test and Brown–Forsythe/Welch ANOVA with Dunnett’s T3 test were used for statistical comparison, *P < 0.05.