Human amnion epithelial cell therapy reduces hypertension-induced vascular stiffening and cognitive impairment

Figures

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  Figure 1. Angiotensin II induced-hypertension and aortic stiffening is reduced by administration of amnion epithelial cells. (A) Immunostaining for HLA-G in angiotensin II and angiotensin II + amnion epithelial cell (AEC) treated mice. Nuclei were stained with haemotoxylin. 400X magnification, scale bar = 50 μm. Representative of n = 3. (B) Effect of amnion epithelial cells (AECs) on angiotensin II-induced hypertension (n = 8–11). All data are mean ± S.E.M. *P < 0.05. Two-way ANOVA with Tukey’s test. (C) Effect of angiotensin II infusion and co-treatment with AECs on pulse wave velocity (n = 6–7). All data are mean ± S.E.M. *P < 0.05. Two-way ANOVA with Sidak’s test. (D) Delta change (day 0 and 14) of pulse wave velocity (n = 6–7). All data are mean ± S.E.M. *P < 0.05. Student’s unpaired t-test.

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  Figure 2. Administration of amnion epithelial cells prevents angiotensin II-induced aortic immune cell infiltration. (A) Representative flow cytometry dot plots showing gating strategy for total leukocytes (CD45+ high) from the aorta of mice infused with vehicle, angiotensin II, vehicle + amnion epithelial cells (AECs) and angiotensin II + AECs. The effect of angiotensin II infusion and co-treatment with AECs on (B) CD45+ leukocytes, (C) CD11b+ myeloid cells, (D) F4/80+ macrophages, (E) Ly6G+ neutrophils, (F) Ly6C+ low monocytes, (G) Ly6C+ high monocytes, (H) CD3+ T cells and (I) CD19+ B cells in the aorta (n = 8–11). All data are mean ± S.E.M. *P < 0.05. Two-way ANOVA with Sidak’s test.

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  Figure 3. Administration of amnion epithelial cells prevents angiotensin II-induced aortic expression of collagen. The effect of angiotensin infusion and co-treatment with amnion epithelial cells (AECs) on mRNA expression of (A) collagen type 1 alpha 1 chain, (B) collagen type 3 alpha 1 chain, (C) collagen type 4 alpha 1 chain and (D) collagen type 5 alpha 1 chain in the aorta (n = 6–8). All data are mean ± S.E.M. *P < 0.05. Two-way ANOVA with Sidak’s test.

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  Figure 4. Administration of amnion epithelial cells improves angiotensin II-induced cognitive impairment. (A) Percentage of time spent in the inner zone in the open field test. (B) Total distance travelled (in cm) in the open field test (C) Representative heatmap plots showing interaction between familiar and novel objects in mice infused with vehicle, angiotensin II, vehicle + amnion epithelilal cells (AECs) and angiotensin II + AECs. (D) Effect of angiotensin II infusion and co-treatment of AECs on recognition memory (n = 7–9). All data are mean ± S.E.M. *P < 0.05. One-sample t-test versus 50%.

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  Figure 5. Administration of amnion epithelial cells modulates genes that are differentially expressed in the brain by angiotensin II-infusion. Volcano plot of differentially expressed genes in brains of mice infused with (A) vehicle versus angiotensin II or (B) angiotensin II versus angiotensin II + AECs. The threshold of differential expression is p-value < 0.05. The horizontal axis is the log2 fold change of genes. The vertical axis is statistical significance scaled as − log 10 p-value. Each dot represents an individual gene (blue: no significant difference; red: upregulated expression; green: downregulated expression). The top upregulated and downregulated genes in brains of mice infused with (C) vehicle versus angiotensin II or (D) angiotensin II versus angiotensin II + AECs (n = 6 per group). Upregulated genes in red and downregulated genes in blue. The colour scale represents the log10 (average FPKM + 1) value.