Effects of Early-Life Stress on Whole-Blood Expression of Vascular Endothelial Dysfunction-Associated Genes

Abstract

Objectives: Early-life stress (ELS) is linked to adult cardiovascular disease, but its immediate effects on adolescent vascular endothelial health remain poorly defined. Here, we evaluated whole-blood expression of vascular endothelial dysfunction-associated genes in ELS-exposed adolescent mice, as well as direct endothelial gene expression changes in dexamethasone-treated human umbilical vein endothelial cells (HUVECs). Methods: BALB/c mice were paired for breeding in a 1:1 male-to-female ratio and allocated into control & experimental groups (6 pairs per group). Offspring from the experimental group underwent maternal separation (3 h /day, postnatal days 1–14) to induce ELS, while control litters remained undisturbed. A total of 24 offspring per group (12 males, 12 females per group) were used for subsequent evaluations. Whole blood from mice was used for gene expression analysis. In parallel, HUVECs were exposed to low-concentration (10 nM) and high-concentration (100 nM) dexamethasone. Target gene expression (Esm1/ESM1, E-selectin/E-SELECTIN, Cst3/CST3, ICAM1, VCAM1) was quantified by Quantitative PCR (qPCR) using the 2-∆∆ct method. Data were analyzed using unpaired t-tests for animal cohort comparisons. Statistical significance was set at p < 0.05. For in vitro HUVECs assays, target gene expression levels were calculated as fold changes relative to vehicle control.   Results: ELS significantly elevated whole-blood Cst3 (p = 0.003) and Esm1 (p = 0.02) expression in adolescent mice across the combined cohort, whereas E-selectin expression remained unaltered. Subgroup analyses revealed a significant transcriptional change in female ELS mice, characterized by an elevation in Cst3 (p = 0.005) and an upward trend in Esm1 (p = 0.060), while male ELS mice showed no significant alterations in any of the targets. Vascular endothelial dysfunction-associated gene expression exhibited concentration-associated differences following dexamethasone exposure. Under low-concentration conditions, CST3 (27.46-fold), ICAM1 (1.46-fold), and VCAM1 (1.41-fold) were upregulated, whereas E-SELECTIN (0.16-fold) and ESM1 (0.42-fold) were downregulated. Under high-concentration conditions, ESM1 (5.83-fold), VCAM1 (4.53-fold), and CST3 (1.61-fold) were upregulated, while ICAM1 (0.49-fold) and E-SELECTIN (0.20-fold) were downregulated. Conclusion: ELS selectively modulated whole-blood expression of vascular endothelial dysfunction-associated genes in adolescent mice, while in the exploratory HUVECs experiment, dexamethasone exposure was associated with distinct gene-expression profiles at 10 nM and 100 nM.  The study identified Cst3 as an exploratory candidate associated with the observed stress response. Future studies in both preclinical models and human adolescent cohorts are needed to confirm its mechanistic role and clinical relevance.