Distinct molecular profiles in human aortic smooth muscle cells under cyclic stretch and hydrostatic pressure

Figures

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  Fig. 1. Overview of transcriptomic analysis of human aortic smooth muscle cells (HASMCs) under different mechanical stress conditions. A: Experimental design. HASMCs were cultured for 24 h under four different mechanical conditions: Ctrl (control, no mechanical stress), S10 (cyclic 10% stretch, 1 Hz), P120 (cyclic normotensive hydrostatic pressure, 120/60 mmHg, 1 Hz), and P180 (cyclic hypertensive hydrostatic pressure 180/120 mmHg, 1 Hz). Total RNA and culture supernatants were collected for bulk RNA-seq and proteomic analysis, respectively. B: Principal component analysis (PCA) of RNA-seq data. Each dot represents one biological replicate (n = 3 per group). C: Hierarchical clustering of RNA-seq data using Ward’s method.

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  Fig. 2. Differential transcriptomic profiles of HASMCs cultured under S10 or P120 conditions. A: Volcano plot of differentially expressed genes (DEGs) identified in the Ctrl vs S10 comparison and heatmap showing the top 20 upregulated and downregulated DEGs. B: Volcano plot of DEGs identified in the Ctrl vs P120 comparison and heatmap showing the top 20 upregulated and downregulated DEGs. Dashed lines in the volcano plot indicate thresholds of false discovery rate (FDR) < 0.05 and |log2FC|= > 1.25. Heatmaps were generated using variance-stabilizing transformed expression values. Genes were ordered by fold change, and samples were arranged according to experimental group. C, D: Gene Ontology (GO) enrichment analysis of upregulated DEGs in the Ctrl vs S10 and Ctrl vs P120 comparisons, respectively. E, F: Venn diagram showing the overlap of upregulated (E) and downregulated (F) DEGs between the S10 and P120 conditions. Numbers in the Venn diagrams indicate the number of DEGs.

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  Fig. 3. Distinct transcriptomic profiles of HASMCs under hypertensive hydrostatic pressure conditions. A: Volcano plot of DEGs identified in Ctrl vs P180 comparison and heatmap showing the top 20 upregulated and downregulated DEGs. B: GO enrichment analysis of upregulated DEGs in the Ctrl vs P180 comparison. C: Venn diagram showing the overlap of DEGs between the S10 and P180 conditions. D: Venn diagram showing the overlap of DEGs between the P120 and P180 conditions. E: qPCR validation of vasoactive intestinal polypeptide receptor 1 (VIPR1) expression under different mechanical stress conditions (n = 7 per group). Individual data points are shown, and horizontal bars indicate mean ± SD. Statistical analysis was performed using the Kruskal–Wallis test followed by pairwise Mann–Whitney U tests.