Figure 3.

Acute mental stress exposure promotes leucocyte recruitment by modulating endothelial cells. (A) Protein levels as mean fluorescent intensities of adhesion molecules expressed by aortic endothelial cells from non-stressed vs. stressed wild-type mice (immediately after 30 min of restraint stress, n = 9–10 per group, Student’s t-test). (B) Pathway analysis of up-regulated genes as assessed by RNA sequencing from fluorescence-activated cell sorting-isolated aortic endothelial cells from non-stressed vs. stressed ApoE^−/− mice (after three episodes of 3 h restraint stress once daily, n = 3 per group). Percentage of up-regulated genes per respective pathway and q-values as assessed by Benjamini–Hochberg correction are shown for the top 10 pathways as defined by P-values. (C) Quantification of plasma chemokine levels measured by ELISA in non-stressed vs. stressed wild-type mice (before, during, and immediately after 3 h of restraint stress, n = 6–18 per group, one-way ANOVA for CCL7, Kruskal–Wallis test for CXCL1, only P-values for comparisons with a control group are annotated). (D) Quantification of stress hormones measured by ELISA in plasma and aortic arches from non-stressed vs. stressed ApoE^−/− mice (immediately after a single 3 h episode of restraint stress, n = 6–14 per group, Student’s t-test, Mann–Whitney U-test for corticosterone). (E–G) Protein levels as mean fluorescent intensities of adhesion molecules expressed by primary murine aortic endothelial cells cultured under high flow conditions (high shear stress, 30 dyn/cm²) incubated with either a stress hormone or respective vehicle [n = 5–8 per group, paired t-test, Wilcoxon test for ICAM1 (all hormones), VCAM1 (corticosterone) and P-selectin (norepinephrine and corticosterone)]. (H–M) Representative images and quantification of monocyte and neutrophil adherence to primary murine aortic endothelial cells cultured under high flow conditions (high shear stress, 30 dyn/cm²) and primed with either a stress hormone or respective vehicle (n = 4 per group, paired t-test). Scale bars represent 100 µm. Data are presented as mean ± SD.