Figure 4.

Role of smooth muscle cells in thrombin generation. (A) ETP values measured at the surface of vascular smooth muscle cells (VSMCs) from LZR and MSZR, with LZR or MSZR platelet free plasma (PFP), and with or without 1.5 g/l exogenous added palmitic acid (PAL). Results are mean ± standard error of the mean, n = 3 with 6 wells per condition per experiment. ^*p < 0.05 vs. LZR VSMC, ^#p < 0.05 vs. LRZ PFP and LRZ VMSC. (B) VSMC associated procoagulant activity reported as phosphatidylserine (PS) equivalent in LRZ and MSZR. Results are mean ± standard error of the mean (n = 25). ^*p < 0.05 vs. LZR. (C) Typical Western blot and (D) quantification analysis of VSMC differentiation markers (αSMA, SM-MHC, and smoothelin) and integrin subunits (α_v and β₃) in cultured VSMCs. Results, expressed as fold change vs. VSMCs from LZR, are mean ± standard error of the mean (n = 6). ^*p < 0.05, MSZR vs. LZR. (E) Representative images of gelatinolytic metalloproteinase activity in the aorta was measured using in situ gelatin zymography for each group of Zucker rats. Fluorescence as marker for intra-plaque gelatinolytic activity was quantified. Nuclei were visualized by DAPI staining. (F) Average wall fluorescence of the gelatinolytic metalloproteinase activity in the aorta. (G) Representative images of zymography gels of LZR and MSZR VSCMCs supernatant at 4, 8, and 20 h. (H) Relative MMP-2 activity in LZR and MSZR VSMC supernatant at 4, 8, and 20 h. Results are mean ± standard error of the mean (n = 5). ^*p < 0.05, MSZR vs. LZR. ELISAs results of MMP-9 (I) and VCAM-1 (J) measured in PFP (n = 17–22). ^*p < 0.05 vs. LZR at the same age; ^#p < 0.05 vs. 25 week-old rats in the same strain.