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Protective effects of hydroxytyrosol-supplemented refined olive oil in animal models of acute inflammation and rheumatoid arthritis S. Silva, B. Sepodes, J. Rocha, R. Direito, A. Fernandes, D. Brites, M. Freitas, E. Fernandes, M.R. Bronze, M.E. Figueira Journal of Nutritional Biochemistry Volume 26, Issue 4, Pages (April 2015) DOI: /j.jnutbio Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 1 Acute inflammation model and impact on rats paw edema (ANOVA, *P<.001 vs. negative control, #P<.001 vs. carrageenan). Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 2 Chronic inflammation model and impact on rats’ paw edema (ANOVA, *P<.001 vs. positive control — RA, +P<.01 vs. ROO). Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 3 Rats’ paws X-rays at day 35 of the RA model; the arrows in the negative control X-ray signalize areas that are prone to arthritic damage, as confirmed in the positive control. Supplemented ROO was supplemented with hydroxytyrosol (5 mg/kg). Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 4 Radiographic score. Scale of 0–3, where 0=no bone damage, 1=tissue swelling and edema, 2=joint erosion and 3=bone erosion and osteophyte formation. **P<.01 vs. positive control — RA. ROO+HT relates to the group receiving hydroxytyrosol-suplemmented (5 mg/kg) ROO. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 5 Histological findings in the collagen-induced arthritic rats: specimen from a vehicle-treated arthritic rat (positive control) exhibits severe synovitis, cartilage damage (large arrow), marked infiltration and pannus (small arrow) and bone resorption (arrow head); specimen from a rat treated with ROO exhibits normal synovium, small cartilage damage (large arrow) and reduced infiltration (small arrow); specimen from a rat treated with hydroxytyrosol-supplemented ROO at 5 mg/kg exhibits normal synovium, no cartilage damage and reduced infiltration (small arrow). Original magnification ×100. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 6 Histological score. Inflammation was scored according to the following criteria: 0=no inflammation; 1=slight thickening of the lining layer or some infiltrating cells in the underlying layer; 2=slight thickening of the lining layer plus some infiltrating cells in the underlying layer; 3=thickening of the lining layer, an influx of cells in the underlying layer and the presence of cells in the synovial space; and 4=synovium highly infiltrated with many inflammatory cells. *P<.01 vs. positive control. ROO+HT relates to the group receiving hydroxytyrosol-suplemmented (5 mg/kg) ROO. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 7 Evaluation of COX-2 and iNOS activation in the collagen-induced arthritic rats after immunostaining of knee joint specimens of arthritic rats for COX-2 (A) and iNOS (B): positive control exhibited marked production of COX-2 and iNOS (brown staining, large arrow); specimens from the treated animals, with ROO and hydroxytyrosol-supplemented ROO at 5 mg/kg, exhibited reduced expression of COX-2 and iNOS. Original magnification ×100. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 8 COX-2 expression quantified using ImageJ software (1.47 v). *P<.01 vs. positive control and **P<.001 vs. positive control. ROO+HT relates to the group receiving hydroxytyrosol-suplemmented (5 mg/kg) ROO. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 9 iNOS expression quantified using ImageJ software (1.47 v). *P<.05 vs. positive control and ***P<.001 vs. positive control. ROO+HT relates to the group receiving hydroxytyrosol-suplemmented (5 mg/kg) ROO. Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 10 Effect of hydroxytyrosol and ROO on the serum levels of TNF-α in the collagen-induced arthritic rats. *P<.005 vs. positive control and+P<.005 vs. ROO. The values are given as the mean±SEM (n=5 per group). Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Fig. 11 Inhibitory effect of hydroxytyrosol (0.25–15.9 μM) on human neutrophils’ oxidative burst stimulated with PMA, as measured by luminol-amplified chemiluminescence. **P<.01 and ***P<.001 compared with the control assay (PMA alone). The values are given as the mean±SEM (n≥4). Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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Journal of Nutritional Biochemistry 2015 26, 360-368DOI: (10. 1016/j
Journal of Nutritional Biochemistry , DOI: ( /j.jnutbio ) Copyright © 2015 Elsevier Inc. Terms and Conditions
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