Hypercoagulability Impairs Plaque Stability in Diabetes-Induced Atherosclerosis

Figure 1. Plasma D-dimer, body weight, blood glucose and plasma lipid levels. (A) Plasma D-dimer levels. (B) Experimental design. Body weight (C), blood glucose levels (D), total plasma cholesterol levels (E), plasma triglyceride levels (F) and plasma HDL levels (G). ApoE-/- and TMPro/Pro ApoE-/- control mice (Cont, normal chow diet, citrate instead of streptozotocin injections), ApoE-/- DM mice and TMPro/Pro ApoE-/- DM (normal chow diet, streptozotocin injections). Each dot represents data obtained from one mouse specimen (n = at least 6 mice per group); ** p < 0.01; ns: non-significant; Mann–Whitney U test (A); two-way ANOVA (C–G).

Figure 2. Smaller plaques in hyperglycemic TMPro/Pro ApoE-/- versus hyperglycemic ApoE-/- mice. (A,B): Representative images of thoracic aortae showing en face plaques as detected by Oil Red O staining (A) and dot-plot summarizing data (total area stained with Oil Red O, (B) in control (normal chow diet, Cont) and diabetic (STZ-induced hyperglycemia, DM) mice. Representative images showing plaques within the aortic arch vessel (C) and dot plot summarizing plaque score (D). ApoE-/- and TMPro/Pro ApoE-/- control mice (Cont, normal chow diet, citrate instead of streptozotocin injections), ApoE-/- DM mice and TMPro/Pro ApoE-/- DM (normal chow diet, streptozotocin injections). Each dot represents data obtained from one mouse specimen (n = at least 6 mice per group); ** p < 0.01; two-way ANOVA.

Figure 3. Less stable plaques in hyperglycemic TMPro/Pro ApoE-/- versus hyperglycemic ApoE-/- mice. (A,B) Representative images showing MOVATs staining (A, lower panel) and Oil Red O staining of brachiocephalic arteries (A, upper panel). Dot plot summarizing the plaque area data (B). Necrotic core area within plaque is indicated by a * and fibrous caps thickness by a black arrow. (C) Dot-plot summarizing lipid deposition within brachiocephalic arteries, Oil Red O-staining. (D,E) Dot plot summarizing the total vessel lumen area (D) and extent of stenosis (E) derived from morphometric analyses of MOVAT-stained images of brachiocephalic arteries. (F,G) Morphometric analyses of MOVATs stained images of brachiocephalic arteries reveal increased necrotic core area (F) and thinner fibrous caps (G), in TMPro/Pro ApoE-/- DM mice compared to ApoE-/- DM. The dot plots in b-f summarize the results obtained from MOVAT-stained brachiocephalic arteries. ApoE-/- and TMPro/Pro ApoE-/- control mice (Cont, normal chow diet, citrate instead of streptozotocin injections), ApoE-/- DM mice and TMPro/Pro ApoE-/- DM (normal chow diet, streptozotocin injections). Each dot represents data obtained from one mouse…

Figure 4. More macrophages and proinflammatory cytokines andfewer smooth muscle cells in hyperglycemic TMPro/Pro ApoE-/- versus hyperglycemic ApoE-/- mice. (A): Representative images showing immunofluorescence staining of macrophages (left panel, MOMA-2, green; DAPI nuclear counterstain, blue) and dot-plot summarizing data (right panel). (B): Representative immunohistochemically stained images showing smooth muscle cells (left panel, α-actin, positive cells detected by HRP-DAB reaction, brown) within lesions and dot-plot summarizing data (right panel). Each dot represents data obtained from one mouse specimen (n = 6, each group); ** p < 0.01, Student’s t-Test.

Figure 5. More proinflammatory cytokines in hyperglycemic TMPro/Pro ApoE-/- versus hyperglycemic ApoE-/- mice. (A–C): Dot plot summarizing data for real-time PCR showing aortic expression of IL-6 (C), IL-1β (D) and TNF-α (E). GAPDH was used as a housekeeping gene. (D–F): Dot plot summarizing data for plasma levels of IL-6 (D), IL-1β (E) and TNF-α (F). ApoE-/- and TMPro/Pro ApoE-/- control mice (Cont, normal chow diet, citrate instead of streptozotocin injections), ApoE-/- DM mice and TMPro/Pro ApoE-/- DM (normal chow diet, streptozotocin injections). Each dot represents data obtained from one mouse specimen (n = at least 6 mice per group); ** p < 0.01, * p < 0.05; ns: non-significant; two-way ANOVA.