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Vasodilatory activity of human quercitin metabolites
P. White et al. / Free Radical Biology and Medicine 75 (2014) S21–S53
fixed. Myeloperoxidase activity was higher in neutrophils of 1-th group patients in comparison with healthy ones. In neutrophils of 2-nd group patients myeloperoxidase activity was higher in comparison with the same of 1 group patients (by 67%, p o0.05). Our results showed the different direction of oxidized proteins formation neutrophils of patients with primary and secondary pneumonia. Besides that the varied degree of myeloperoxidase activity was fixed. Our results require more detailed understanding because they can reflect peculiar mechanisms of pneumonia development and determine the characteristics of their progression.
http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.796
P65
The role of HSP90 in Quercetin-induced apoptosis in human papillary thyroid (B-CPAP) cancer cells Ergul Mutlu Altundag a,b, Ayse Mine Yilmaz a,b, Tolga Kasaci a, Ceyda Corek a,b, Yavuz Taga a,b, A. Suha Yalçin a,b a
Department of Biochemistry, School of Medicine, Marmara University, Istanbul, Turkey b Genetic and Metabolic Diseases Research and Investigation Center, Marmara University, Istanbul, Turkey
S43
Abstract A large number of studies confirmed antihypertensive effect of quercetin. However, due to its low bioavailability, the mechanism of action has been remaining vague and is not likely associated with its antioxidant effect. This study was designed to determinate if quercetin metabolites might be responsible for the activity. During the in vitro study, the vasorelaxant potency of different quercetin metabolites including small phenolic acids formed by bacterial microflora was tested. Metabolites at concentration 10-7 to 10-3 M were tested for relaxation of rat thoracic aorta rings precontracted by phenylephrine. Subsequently the most active structure was selected for in vivo experiments, when the effect on blood pressure and heart rate was monitored after i.v. administration in dose range from 0,2 mg/kg to 25 mg/kg. The most active structure, 3-(3-hydroxyphenyl)propionic acid, initiated vasorelaxation in concentration of 100 nM while quercetin at 500 nM. The major quercetin metabolite formed by human enzymes, 3-glucuronide, was almost inactive. During the in vivo study the 3-(3-hydroxyphenyl)propionic acid decreased systolic blood pressure even at dose of 1 mg/kg without having an effect on heart rate at this dose. In conclusion, some of quercetin metabolites may have be partly responsible for vasorelaxant activity of orally administered quercetin.
Acknowledgments: This work was financed by the grant P303/ 12/G163 of the Czech Science Foundation and by the grant 170/50/ 25003 of Charles University. http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.798
Abstract As in many other cancers, genetic alterations that occur in genes encoding for key proteins of major signalling pathways are the driving force for the tumorigenesis of thyroid cancer. HSP90 is an emerging therapeutic target of interest for the treatment of cancer and is responsible for modulating cellular response to stress by maintaining the function of signalling proteins. In this study, we have worked with B-CPAP, a thyroid papillary cancer cell line which is known to have BRAF V600E mutation. We have administered the flavonoid Quercetin, which is known to induce apoptosis by inhibiting HSP production on various cancer cell lines, at different concentrations (between 10 and 75 mM) for 24 hours. We have measured cell viability using WST-1 assay. We found that the viability decreases in a dose dependent manner. Then, we chose Quercetin concentrations between 10-75 mM for 24 hours, for the apoptosis and cell cycle analysis in flow cytometry by Annexin V and propidium iodide. We have also applied Hoechst stain to cancer cells for visualizing them on fluorescence microscopy and confirmed apoptosis with the presence of apoptotic signs in nuclei of cancer cells. Finally, we used Western blotting to compare HSP90 and Cleaved-PARP levels in cells treated with Quercetin and the control group. In the light of the obtained data, our results suggest that in future studies it would be useful to investigate the apoptotic mechanisms of Quercetin and use combinational therapies on BCPAP cells. Supported by Marmara University Scientific Research Commission (SAG-C-TUP-130313-0063).
http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.797
Food and Nutrition Poster Prize Winner P66
Vasodilatory activity of human quercitin metabolites Najmanová Iveta, Vopršálová Marie, Mladěnka Přemysl Charles University in Prague (Faculty of Pharmacy in Hradec Králové), Department of Pharmacology and Toxicology, Czech Republic
P67
Redox regulation of E3 ubiquitin ligases and their role in skeletal muscle atrophy Olaso-Gonzalez Gloriaa, Ferrando Beatriza, Derbre Fredericb, Salvador-Pascual Andreaa, Cabo Helenaa, Pareja-Galeano Heliosa, Sabater-Pastor Frederica, Gomez-Cabrera Mari Carmena, Vina Josea a
University of Valencia (Faculty of Medicine), Department of Physiology, Spain b University of Rennes2 (UFR APS), Laboratory « Movement, Sport and health Sciences, France
Abstract Muscle atrophy is linked to reactive oxygen species (ROS) production during hindlimb-unloading due, at least in part, to the activation of xanthine oxidase (XO). The major aim of our study was to determine the mechanism by which ROS cause muscle atrophy and its possible prevention by allopurinol, a well-known inhibitor of XO widely used in clinical practice, and indomethacin, a nonsteroidal anti-inflammatory drug. We studied the activation of p38 MAP Kinase and NF-κB pathways, and the expression of two E3 ubiquitin ligases involved in proteolysis, the Muscle atrophy F-Box (MAFb) and Muscle RING Finger-1 (MuRF-1). Male Wistar rats (3 mold) conditioned by 14 days of hindlimb unloading (n¼18), with or without the treatment, were compared with freely ambulating controls (n¼18). After the experimental intervention, soleus muscles were removed, weighted and analyzed to determine oxidative stress and inflammatory parameters. We found that hindlimb unloading induced a significant increase in XO activity in plasma (39%, p¼0.001) and in the protein expression of CuZnSOD and Catalase in skeletal muscle. Inhibitionof XO partially prevented protein carbonylation, both in plasma and in soleus muscle, in the unloaded animals. The most relevant new fact reported is that allopurinol prevents soleus muscle atrophy by 20% after hindlimb unloading. Combining allopurinol and indomethacin we found a further prevention in the atrophy process.
fixed. Myeloperoxidase activity was higher in neutrophils of 1-th group patients in comparison with healthy ones. In neutrophils of 2-nd group patients myeloperoxidase activity was higher in comparison with the same of 1 group patients (by 67%, p o0.05). Our results showed the different direction of oxidized proteins formation neutrophils of patients with primary and secondary pneumonia. Besides that the varied degree of myeloperoxidase activity was fixed. Our results require more detailed understanding because they can reflect peculiar mechanisms of pneumonia development and determine the characteristics of their progression.
http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.796
P65
The role of HSP90 in Quercetin-induced apoptosis in human papillary thyroid (B-CPAP) cancer cells Ergul Mutlu Altundag a,b, Ayse Mine Yilmaz a,b, Tolga Kasaci a, Ceyda Corek a,b, Yavuz Taga a,b, A. Suha Yalçin a,b a
Department of Biochemistry, School of Medicine, Marmara University, Istanbul, Turkey b Genetic and Metabolic Diseases Research and Investigation Center, Marmara University, Istanbul, Turkey
S43
Abstract A large number of studies confirmed antihypertensive effect of quercetin. However, due to its low bioavailability, the mechanism of action has been remaining vague and is not likely associated with its antioxidant effect. This study was designed to determinate if quercetin metabolites might be responsible for the activity. During the in vitro study, the vasorelaxant potency of different quercetin metabolites including small phenolic acids formed by bacterial microflora was tested. Metabolites at concentration 10-7 to 10-3 M were tested for relaxation of rat thoracic aorta rings precontracted by phenylephrine. Subsequently the most active structure was selected for in vivo experiments, when the effect on blood pressure and heart rate was monitored after i.v. administration in dose range from 0,2 mg/kg to 25 mg/kg. The most active structure, 3-(3-hydroxyphenyl)propionic acid, initiated vasorelaxation in concentration of 100 nM while quercetin at 500 nM. The major quercetin metabolite formed by human enzymes, 3-glucuronide, was almost inactive. During the in vivo study the 3-(3-hydroxyphenyl)propionic acid decreased systolic blood pressure even at dose of 1 mg/kg without having an effect on heart rate at this dose. In conclusion, some of quercetin metabolites may have be partly responsible for vasorelaxant activity of orally administered quercetin.
Acknowledgments: This work was financed by the grant P303/ 12/G163 of the Czech Science Foundation and by the grant 170/50/ 25003 of Charles University. http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.798
Abstract As in many other cancers, genetic alterations that occur in genes encoding for key proteins of major signalling pathways are the driving force for the tumorigenesis of thyroid cancer. HSP90 is an emerging therapeutic target of interest for the treatment of cancer and is responsible for modulating cellular response to stress by maintaining the function of signalling proteins. In this study, we have worked with B-CPAP, a thyroid papillary cancer cell line which is known to have BRAF V600E mutation. We have administered the flavonoid Quercetin, which is known to induce apoptosis by inhibiting HSP production on various cancer cell lines, at different concentrations (between 10 and 75 mM) for 24 hours. We have measured cell viability using WST-1 assay. We found that the viability decreases in a dose dependent manner. Then, we chose Quercetin concentrations between 10-75 mM for 24 hours, for the apoptosis and cell cycle analysis in flow cytometry by Annexin V and propidium iodide. We have also applied Hoechst stain to cancer cells for visualizing them on fluorescence microscopy and confirmed apoptosis with the presence of apoptotic signs in nuclei of cancer cells. Finally, we used Western blotting to compare HSP90 and Cleaved-PARP levels in cells treated with Quercetin and the control group. In the light of the obtained data, our results suggest that in future studies it would be useful to investigate the apoptotic mechanisms of Quercetin and use combinational therapies on BCPAP cells. Supported by Marmara University Scientific Research Commission (SAG-C-TUP-130313-0063).
http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.797
Food and Nutrition Poster Prize Winner P66
Vasodilatory activity of human quercitin metabolites Najmanová Iveta, Vopršálová Marie, Mladěnka Přemysl Charles University in Prague (Faculty of Pharmacy in Hradec Králové), Department of Pharmacology and Toxicology, Czech Republic
P67
Redox regulation of E3 ubiquitin ligases and their role in skeletal muscle atrophy Olaso-Gonzalez Gloriaa, Ferrando Beatriza, Derbre Fredericb, Salvador-Pascual Andreaa, Cabo Helenaa, Pareja-Galeano Heliosa, Sabater-Pastor Frederica, Gomez-Cabrera Mari Carmena, Vina Josea a
University of Valencia (Faculty of Medicine), Department of Physiology, Spain b University of Rennes2 (UFR APS), Laboratory « Movement, Sport and health Sciences, France
Abstract Muscle atrophy is linked to reactive oxygen species (ROS) production during hindlimb-unloading due, at least in part, to the activation of xanthine oxidase (XO). The major aim of our study was to determine the mechanism by which ROS cause muscle atrophy and its possible prevention by allopurinol, a well-known inhibitor of XO widely used in clinical practice, and indomethacin, a nonsteroidal anti-inflammatory drug. We studied the activation of p38 MAP Kinase and NF-κB pathways, and the expression of two E3 ubiquitin ligases involved in proteolysis, the Muscle atrophy F-Box (MAFb) and Muscle RING Finger-1 (MuRF-1). Male Wistar rats (3 mold) conditioned by 14 days of hindlimb unloading (n¼18), with or without the treatment, were compared with freely ambulating controls (n¼18). After the experimental intervention, soleus muscles were removed, weighted and analyzed to determine oxidative stress and inflammatory parameters. We found that hindlimb unloading induced a significant increase in XO activity in plasma (39%, p¼0.001) and in the protein expression of CuZnSOD and Catalase in skeletal muscle. Inhibitionof XO partially prevented protein carbonylation, both in plasma and in soleus muscle, in the unloaded animals. The most relevant new fact reported is that allopurinol prevents soleus muscle atrophy by 20% after hindlimb unloading. Combining allopurinol and indomethacin we found a further prevention in the atrophy process.