Unveiling the molecular Culprit of arterial stiffness in vitamin D deficiency and obesity: Potential for novel therapeutic targets

Figures

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  Fig. 1. Pulse wave velocity relative to age in vitamin D deficiency and obese patients (A-group), n = 9. compared to control healthy individuals with normal Vitamin D level and BMI (C-group), n = 11. Values are expressed as mean ± SEM and analyzed using unpaired two-tailed Student t-test. *P ≤ 0.05.

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  Fig. 2. Overall pipeline of RNA sequencing and bioinformatics analysis.

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  Fig. 3. Volcano plot showing the DEGs between the vitamin D deficiency and obese group compared to the healthy control group. The red scatters present the upregulated genes, bleu scatters present the downregulated genes and grey scatters indicate no significant differentially expressed genes between the vitamin D deficiency and obese group compared to the healthy control group. The top significantly expressed genes (FDR <0.05) between the vitamin D deficiency and obese group compared to the healthy control group were labeled.

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  Fig. 4. Functional annotation and enrichment analysis of the top differentially expressed genes resulting in Vit D deficiency and obese group compared to the controls group. The top significant enriched pathways are displayed (Benjamini-Hochberg adjusted p < 0.05).

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  Fig. 5. Gene expression levels of HIF-1 alpha and NADPH Oxidase: HIF-1 alpha (A), NOX-I (B), NOX-II (C), and NOX-V (D) using qRT-PCR analysis in patients with vitamin D deficiency and obese (A-group) compared to healthy individuals with normal level of Vit. D and BMI, control group (C-group). Calculated mean ± SD fold change in gene expression levels in patient's plasma based on two separate experiments. *P ≤ 0.05, **P ≤ 0.01, determined using unpaired two-tailed Student t-test.

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  Fig. 6. Western blot analysis showing HIF-1 alpha and NADPH Oxidase protein expression: HIF-1 alpha (A), NOX-I (B), NOX-II (C), and NOX-V (D) in patients with vitamin D deficiency and obese (A-group) compared to healthy individuals with normal level of Vit. D and BMI, control group (C-group). Calculated mean ± SD fold change in protein-expression levels in patient's plasma based on two separate experiments. *P ≤ 0.05, **P ≤ 0.01, determined using unpaired two-tailed Student t-test. Note that full uncropped images are provided in the supplementary materials.

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  Fig. 7. Relative gene expressions of inflammatory and angiogenesis markers A-D (Il-1b, IL-8, IL-10, and VEGF) in patients with vitamin D deficiency and obese (A-group) compared to healthy individuals with normal level of Vit. D and BMI, control group (C-group). Values are expressed as mean ± SEM; (n = 9). Analyzed using unpaired two-tailed Student t-test. *P ≤ 0.05, **P ≤ 0.01 ***P ≤ 0.001.

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  Fig. 8. Relative proteins expressions of inflammatory and angiogenesis markers A-D (Il-1b, IL-8, IL-10, and VEGF) in patients with vitamin D deficiency and obese (A-group) compared to healthy individuals with normal level of Vit. D and BMI, control group (C-group). Values are expressed as mean ± SEM; (n = 9–11). Analyzed using unpaired two-tailed Student t-test. *P ≤ 0.05, **P ≤ 0.01.