Differences in Distribution and Biological Effects of F3O4@PEG Nanoparticles in Normotensive and Hypertensive Rats—Focus on Vascular Function and Liver

Figures

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  Figure 1. An example of the original recording of noradrenaline—(a) and serotonin-induced (b) contractions and acetylcholine-induced endothelium-dependent relaxations, followed by spontaneous endothelium-dependent contractions in the femoral arteries of control and USPION-treated rats. Abbreviations: ACh, acetylcholine; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats; NA, noradrenaline; 5-HT, 5-hydroxytryptamine (serotonin).

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  Figure 2. Total distance travelled (a), relative distance travelled in the central zone (b), time spent in the central zone (c), and central entries in the open-field test (d). Results are mean ± SEM. See the Results section for detailed 3-way ANOVA of results. Abbreviations: USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 3. Blood pressure (a), relative blood pressure changes (b), heart rate (c), and relative heart rate (d) in rats. Results are mean ± SEM. See the Results section for detailed ANOVA results. + p < 0.05 vs. basal, * p < 0.05 vs. SHRc, x p < 0.05 vs. WKYu. Abbreviations: USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 4. Nitric oxide synthase activity in the brainstem. Results are mean ± SEM. Abbreviations: NOS, nitric oxide synthase activity; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 5. Nitric oxide synthase activity in the aorta (a), eNOS gene expression (b) iNOS gene expression (c), correlation between eNOS and iNOS gene expression (d), correlation between eNOS gene expression and total NOS activity (e), and correlation between iNOS gene expression and total NOS activity (f). Results are mean ± SEM. See the Results section for detailed 2-way ANOVA results. Abbreviations: eNOS, endothelial nitric oxide synthase; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 6. Nitric oxide synthase activity in the liver (a), eNOS gene expression (b) iNOS gene expression (c), correlation between eNOS and iNOS gene expression (d), correlation between eNOS gene expression and total NOS activity (e), and correlation between iNOS gene expression and total NOS activity (f). Results are mean ± SEM. See the Results section for detailed 2-way ANOVA results. * p < 0.05 vs. SHRc, x p < 0.05 vs. the respective WKY group. Abbreviations: eNOS, endothelial nitric oxide synthase; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 7. Gene expressions of SOD1 (a), SOD2 (b), NRF2 (c), PPARγ (d), DMT1 (e), and TfR1 (f) genes in the aorta. Results are mean ± SEM. See the Results section for detailed 2-way ANOVA results. Abbreviations: NRF2, nuclear factor erythroid 2–related factor 2; PPARγ, peroxisome proliferator-activated receptor gamma; SOD1, superoxide dismutase 1; SOD2, superoxide dismutase 2; DMT1, divalent metal transporter 1; TfR1, transferrin receptor 1; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 8. Gene expression of SOD1 (a), SOD2 (b), NRF2 (c), PPARγ (d), DMT1 (e) TfR1 (f), HAMP (g), and FTH1 (h) in the liver. Results are mean ± SEM. See the Results section for detailed 2-way ANOVA results. * p < 0.05 vs. SHRc, x p < 0.05 vs. WKYu. Abbreviations: NRF2, nuclear factor erythroid 2–related factor 2; PPARγ, peroxisome proliferator-activated receptor gamma; SOD1, superoxide dismutase 1; SOD2, superoxide dismutase 2; DMT1, divalent metal transporter 1; TfR1, transferrin receptor 1; HAMP, hepcidin; FTH1, ferritin H1 subunit; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 9. 5-hydroxytryptamine—(a) and noradrenaline-induced (b) contractions in the femoral arteries in the control and USPION-treated rats. Abbreviations: WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats; NA, noradrenaline; 5-HT, 5-hydroxytryptamine (serotonin).

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  Figure 10. Acetylcholine-induced endothelium-dependent relaxation (a) and acetylcholine-induced endothelium-dependent contractions (b) in the femoral arteries. The values represent the mean ± SEM. See the Results section for detailed 2-way ANOVA results. * p < 0.02 SHRc vs. SHRu. Abbreviations: ACh, acetylcholine; 5-HT, 5-hydroxytryptamine; USPIONs, ultra-small superparamagnetic iron oxide nanoparticles; WKYc, control Wistar–Kyoto rats; WKYu, USPION-treated Wistar–Kyoto rats; SHRc, control spontaneously hypertensive rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 11. USPION-originated iron content in urine (a), faeces (b), blood (c), serum (d), aorta (e), and liver (f). The values represent the mean ± SEM. x p < 0.05 vs. WKYu, n = 5–6/group. Abbreviations: WKYu, USPION-treated Wistar–Kyoto rats; SHRu, USPION-treated spontaneously hypertensive rats.

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  Figure 12. Representative electron micrographs of the unstained ultra-thin sections of the aorta of USPION-treated Wistar–Kyoto rats (a,b) demonstrating distribution of nanoparticles (solid arrows) within the elastic layers. In USPION-treated spontaneously hypertensive rats, nanoparticles were present in the smooth muscle cells (dashed arrows), as well as within the elastic layers (solid arrows) (c,d). In the liver, a heterogeneous distribution of nanoparticles was observed in hepatocytes of WKY (e) and SHR (f). Original magnifications: ×4000. Abbreviations: smc, vascular smooth muscle cells; m, mitochondria.