Role of Lectin-like Oxidized LDL Receptor-1 and Syncytiotrophoblast Extracellular Vesicles in the Vascular Reactivity of Mouse Uterine Arteries During Pregnancy

Figures

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  Figure 1. Nitric oxide contribution to endothelium-dependent vasodilation in uterine arteries from WT and LOX-1tg mice. (a,c) Contribution of nitric oxide to endothelium-dependent vasodilation responses to increasing doses of methylcholine (MCh) in uterine arteries from WT (a,b) and LOX-1tg mice (c,d) incubated overnight with (blue lines) or without STBEVs (black lines). Nitric oxide contribution to vasodilation was assessed by pre-incubation with (dashed lines; triangles) or without (solid lines; squares) the pan nitric oxide synthase inhibitor L-NAME. (b,d) Summary graphs show maximal vasodilation responses to MCh (Emax) in uterine arteries from WT (b) and LOX-1tg mice (d) incubated overnight with (blue bars) or without STBEVs (white bars). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; n.s.: not significant; ##p < 0.01, ####p < 0.0001, overall effect of treatment (control vs. L-NAME) in two-way ANOVA; **p < 0.01, control vs. L-NAME in multiple comparisons post-test. n = 8–10/group.

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  Figure 2. Effect of oxLDL on endothelium-dependent vasodilation in uterine arteries from WT and LOX-1tg mice. (a,c) Contribution of pre-incubation with (dashed lines; circles) or without (solid lines; squares) oxLDL (oxLDL-induced LOX-1 activation; 10 minutes before the start of the curve) to endothelium-dependent vasodilation responses to increasing doses of methylcholine (MCh) in uterine arteries from WT (a,b) and LOX-1tg mice (c,d) incubated overnight with (blue lines) or without STBEVs (black lines). (b,d) Summary graphs show the dose of to MCh necessary to establish 50% of total relaxation (pEC50) in uterine arteries from WT (b) and LOX-1tg mice (d) incubated overnight with (blue bars) or without STBEVs (white bars). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; n.s.: not significant; ##p < 0.01, overall effect of treatment (control vs. oxLDL) in two-way ANOVA; *p < 0.05, control vs. oxLDL in multiple comparisons post-test. n = 8–9/group.

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  Figure 3. Ang II-induced vasoconstriction responses in uterine arteries from WT and LOX-1tg mice. (a) Vascular responses to increasing doses of Ang II in uterine arteries from WT (black lines) and LOX-1tg mice (red lines) incubated overnight with (dashed lines; triangles) or without STBEVs (solid lines; squares). (b) Summary graph show area under the curve (AUC) in uterine arteries from WT (white bars) and LOX-1tg mice (red bars) incubated overnight with (triangles) or without STBEVs (squares). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; n.s.: not significant; †p < 0.05 overall effect of strain (WT vs. LOX-1tg) in two-way ANOVA; *p < 0.05, control vs. STBEVs in multiple comparisons post-test. n = 9–10/group.

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  Figure 4. AT1 receptor contribution to Ang II-induced vasoconstriction responses in uterine arteries from WT and LOX-1tg mice. (a,b) The contribution of the AT1 receptor on the Ang II-induced vasoconstriction responses in uterine arteries from WT (a) and LOX-1tg mice (b) incubated overnight with (blue lines) or without STBEVs (black lines) was assessed by pre-incubation with (dashed lines; diamonds) or without (solid lines; squares) the AT1 receptor antagonist candesartan. (c) Summary graph shows the delta area under the curve (ΔAUC) between the control and candesartan-incubated uterine arteries from WT (white bars) and LOX-1tg mice (red bars) incubated overnight with (triangles) or without STBEVs (squares). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; n.s.: not significant; †p < 0.05 overall effect of strain (WT vs. LOX-1tg); ##p < 0.01, overall effect of treatment (control vs. STBEVs) in two-way ANOVA; **p < 0.01, control vs. STBEVs in multiple comparisons post-test. n = 7–10/group.

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  Figure 5. Uterine artery AT1 receptor expression. (a) Using immunofluorescent staining, AT1 receptor expression was assessed in cryo-sections of uterine arteries from WT (white bars) and LOX-1tg mice (red bars) incubated overnight with (triangles) or without STBEVs (squares). (b) Representative uterine artery images for each of the groups (red = AT1 and blue=nuclear dapi staining). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; #p < 0.05, overall effect of treatment (control vs. STBEVs) in two-way ANOVA; †p < 0.05, overall effect of strain (WT vs. LOX-1tg) in two-way ANOVA; **p < 0.01, control vs. STBEVs in multiple comparisons post-test. n = 5–6/group.

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  Figure 6. Effect of oxLDL on Ang II-induced vasoconstriction responses. (a,b) Contribution of pre-incubation with (dashed lines; circles) or without (solid lines; squares) oxLDL (10 minutes before the start of the curve) to Ang II-induced vasoconstriction responses in uterine arteries from WT (a) and LOX-1tg mice (b) incubated overnight with (blue lines) or without STBEVs (black lines). (c) Summary graph shows delta (Δ) response to Ang II at the highest Ang II dose between the control and oxLDL-incubated uterine arteries from WT (white bars) and LOX-1tg mice (red bars) incubated overnight with (triangles) or without STBEVs (squares). Means with SEM; two-way ANOVA with Sidak’s multiple comparisons post-test; n.s.: not significant; ####p < 0.0001, overall effect of treatment (control vs. STBEVs) in two-way ANOVA; *p < 0.05, **p < 0.01, control vs. STBEVs in multiple comparisons post-test. n = 8–9/group.