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Effect of vitamin D3 supplementation on metal and redox homeostasis in hormone treated prostate cancer patients
A. Russo et al. / Free Radical Biology and Medicine 96 (2016) S15–S20
observed, consistent with interaction of the enzyme with fibronectin and site-specific oxidant formation. A number of the modifications identified with both oxidant systems were mapped to functional domains on the protein. These include reversible and irreversible oxidation of Cys-135 in the fibrin I/heparin I binding domain, oxidation of Met-1283 in the cell-binding domain, oxidation of Met-2224 in the fibrin II binding domain, and irreversible oxidation of Cys-374 in the collagen-binding domain. Conclusions: The observed modifications are consistent with oxidation by the myeloperoxidase/HOCl system inducing significant changes in fibronectin structure and function, which may modulate ECM assembly and integrity, including the capacity to support cell adhesion. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.157 YI-10
Effect of vitamin D3 supplementation on metal and redox homeostasis in hormone treated prostate cancer patients Krisztina Süle 1,2, Klára Szentmihályi 1,2, Gergő Szabó 1, Dénes Kleiner 1, Imre Varga 3, Anna Egresi 1, Zoltán May 2, Péter Nyirády 4, Anna Blázovics 1 1
Institute of Farmacognosy, Semmelweis University, Budapest, Hungary 2 Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, Hungary 3 Medical Center of Dunakeszi, Dunakeszi, Hungary 4 Department of Urology and Urooncological Centre, Semmelweis University, Budapest, Hungary
Prostate cancer is one of the most common malignant diseases in Europe. Metal element status could be a good prognostic index in prostate adenocarcinoma, therefore the aim of our investigation was to demonstrate the effect of vitamin-D3 consumption on metal-homeostasis. Methods: We examined the metal element status in erythrocytes of healthy and prostate cancer patients with total androgen blockade with and without vitamin D3 supplementation. Forty three volunteers were divided into 5 groups: healthy (N ¼8); precancerous (N ¼9); healthy with vitamin D3(N ¼ 6); precancerous with vitamin D3(N ¼8); neoplasma prostata with vitamin D3(N ¼11). The micro and macro element (20 elements) concentrations of erythrocytes were determined with ICP-OES. Permission: IKEB 01/2015. Results: Vitamin D3 supplementation had positive effect on the concentrations of Mn, Cu, Fe, Mg, Cr, Ca, Ni in erythrocytes of prostate cancer patients. In some cases, concentration of P, B, Ba, Sr, Zn were slightly altered by vitamin D3 consumption negatively. Conclusion: Vitamin D3 supplementation affected in a positive way the outcome of the disease that does not only appear in the clinical laboratory results or blood redox parameters but the metal
S19
element status. However, further comprehensive investigations are required to understand the role of vitamin D3 on element status of prostate cancer patients. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.158 YI-11
α-130 -OH is the main product of α-tocopherol metabolism and influences CYP4F2 and PPARγ gene expression in HepG2 human hepatocarcinoma cells Pierangelo Torquato 1, Desirée Bartolini 1, Danilo Giusepponi 2, Giorgio Saluti 2, Angelo Russo 1, Carolina Barola 1, Marc Birringer 3, Roberta Galarini 2, Francesco Galli 1 1
Dept. of Pharmaceutical Sciences, University of Perugia, Perugia, Italy 2 Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, Perugia, Italy 3 University of Applied Sciences Fulda, Fulda, Hessen, Germany
Cytochrome P-450 mediated ω-oxidation is the earliest step in the enzymatic metabolism of vitamin E (tocopherols and tocotrienols). It generates the long chain carboxychromanol metabolites (α-LCMs) α-13’-OH and its corresponding carboxylic acid α13’-COOH. These have been demonstrated to be active derivatives of vitamin E metabolism with in vitro effects on inflammatory and lipid metabolism pathways. Since an unbiased analysis procedure has been developed in our laboratories to measure such LCMs in human blood and tissues (Ciffolilli et al. FRBM 2015; Torquato et al. JPBA 2016), in this study we aim at provide a first quantitative characterization of vitamin E metabolism in human hepatocites supplemented with vitamin E and to explore if these metabolites may have themselves a biological role in the regulation of CYP4F2 gene, the main isoenzyme proposed to have tocopherol ω-oxidase activity, and of other genes at the interface with lipid metabolism and inflammation such as the nuclear receptor PPARγ. When supplemented with a bolus of RRR-α-tocopherol (25 or 100 mM), HepG2 cells showed a significant up-regulation of CYP4F2 protein and then generated both the α-LCMs. At different time points in this in vitro observation, α-13’-OH was recovered to a much greater extent than α-13’-COOH in the culture medium being almost the exclusive form measured at 24 hrs post-supplementation. Preliminary quantification data suggest that 1 x 106 HepG2 cells supplemented with 25 mM RRR-α-tocopherol approximately produce in 24 hrs from 50 to 160 pmol of α-13’OH, which is indicative of a robust and steady metabolism of this vitamin. α-13’-COOH increased progressively to reach a molar ratio over α-13’-OH of 1/30 at 48 hrs, 1/14 at 72 hr and then of 1/16 at 96 hours. These findings demonstrate that the first step of ω-oxidation is not supported by a rapid conversion by aldehyde or alcohol dehydrogenase enzymes to α-13’-COOH for further degradation to short-chain metabolites. As a consequence, α-13’-OH is the main LCM formed during HepG2 metabolism of RRR-α-tocopherol and significant intracellular concentrations (in the low micromolar range) of this LCM are expected to form during supplementation.
observed, consistent with interaction of the enzyme with fibronectin and site-specific oxidant formation. A number of the modifications identified with both oxidant systems were mapped to functional domains on the protein. These include reversible and irreversible oxidation of Cys-135 in the fibrin I/heparin I binding domain, oxidation of Met-1283 in the cell-binding domain, oxidation of Met-2224 in the fibrin II binding domain, and irreversible oxidation of Cys-374 in the collagen-binding domain. Conclusions: The observed modifications are consistent with oxidation by the myeloperoxidase/HOCl system inducing significant changes in fibronectin structure and function, which may modulate ECM assembly and integrity, including the capacity to support cell adhesion. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.157 YI-10
Effect of vitamin D3 supplementation on metal and redox homeostasis in hormone treated prostate cancer patients Krisztina Süle 1,2, Klára Szentmihályi 1,2, Gergő Szabó 1, Dénes Kleiner 1, Imre Varga 3, Anna Egresi 1, Zoltán May 2, Péter Nyirády 4, Anna Blázovics 1 1
Institute of Farmacognosy, Semmelweis University, Budapest, Hungary 2 Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, Hungary 3 Medical Center of Dunakeszi, Dunakeszi, Hungary 4 Department of Urology and Urooncological Centre, Semmelweis University, Budapest, Hungary
Prostate cancer is one of the most common malignant diseases in Europe. Metal element status could be a good prognostic index in prostate adenocarcinoma, therefore the aim of our investigation was to demonstrate the effect of vitamin-D3 consumption on metal-homeostasis. Methods: We examined the metal element status in erythrocytes of healthy and prostate cancer patients with total androgen blockade with and without vitamin D3 supplementation. Forty three volunteers were divided into 5 groups: healthy (N ¼8); precancerous (N ¼9); healthy with vitamin D3(N ¼ 6); precancerous with vitamin D3(N ¼8); neoplasma prostata with vitamin D3(N ¼11). The micro and macro element (20 elements) concentrations of erythrocytes were determined with ICP-OES. Permission: IKEB 01/2015. Results: Vitamin D3 supplementation had positive effect on the concentrations of Mn, Cu, Fe, Mg, Cr, Ca, Ni in erythrocytes of prostate cancer patients. In some cases, concentration of P, B, Ba, Sr, Zn were slightly altered by vitamin D3 consumption negatively. Conclusion: Vitamin D3 supplementation affected in a positive way the outcome of the disease that does not only appear in the clinical laboratory results or blood redox parameters but the metal
S19
element status. However, further comprehensive investigations are required to understand the role of vitamin D3 on element status of prostate cancer patients. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.158 YI-11
α-130 -OH is the main product of α-tocopherol metabolism and influences CYP4F2 and PPARγ gene expression in HepG2 human hepatocarcinoma cells Pierangelo Torquato 1, Desirée Bartolini 1, Danilo Giusepponi 2, Giorgio Saluti 2, Angelo Russo 1, Carolina Barola 1, Marc Birringer 3, Roberta Galarini 2, Francesco Galli 1 1
Dept. of Pharmaceutical Sciences, University of Perugia, Perugia, Italy 2 Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, Perugia, Italy 3 University of Applied Sciences Fulda, Fulda, Hessen, Germany
Cytochrome P-450 mediated ω-oxidation is the earliest step in the enzymatic metabolism of vitamin E (tocopherols and tocotrienols). It generates the long chain carboxychromanol metabolites (α-LCMs) α-13’-OH and its corresponding carboxylic acid α13’-COOH. These have been demonstrated to be active derivatives of vitamin E metabolism with in vitro effects on inflammatory and lipid metabolism pathways. Since an unbiased analysis procedure has been developed in our laboratories to measure such LCMs in human blood and tissues (Ciffolilli et al. FRBM 2015; Torquato et al. JPBA 2016), in this study we aim at provide a first quantitative characterization of vitamin E metabolism in human hepatocites supplemented with vitamin E and to explore if these metabolites may have themselves a biological role in the regulation of CYP4F2 gene, the main isoenzyme proposed to have tocopherol ω-oxidase activity, and of other genes at the interface with lipid metabolism and inflammation such as the nuclear receptor PPARγ. When supplemented with a bolus of RRR-α-tocopherol (25 or 100 mM), HepG2 cells showed a significant up-regulation of CYP4F2 protein and then generated both the α-LCMs. At different time points in this in vitro observation, α-13’-OH was recovered to a much greater extent than α-13’-COOH in the culture medium being almost the exclusive form measured at 24 hrs post-supplementation. Preliminary quantification data suggest that 1 x 106 HepG2 cells supplemented with 25 mM RRR-α-tocopherol approximately produce in 24 hrs from 50 to 160 pmol of α-13’OH, which is indicative of a robust and steady metabolism of this vitamin. α-13’-COOH increased progressively to reach a molar ratio over α-13’-OH of 1/30 at 48 hrs, 1/14 at 72 hr and then of 1/16 at 96 hours. These findings demonstrate that the first step of ω-oxidation is not supported by a rapid conversion by aldehyde or alcohol dehydrogenase enzymes to α-13’-COOH for further degradation to short-chain metabolites. As a consequence, α-13’-OH is the main LCM formed during HepG2 metabolism of RRR-α-tocopherol and significant intracellular concentrations (in the low micromolar range) of this LCM are expected to form during supplementation.