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Oxidative stress and Na+-glucose cotransporters Sglt1 and Sglt2 in kidneys of ochratoxin A-treated rats
Abstracts / Toxicology Letters 205S (2011) S180–S300
caspase 3 and glutathione S-transferase were assessed using standard protocols. Results: We found significant depletion of liver and brain low molecular weight antioxidants. On the other hand, the depletion was partially recovered by an increase in the GSH level. Obidoxime was implicated in alteration of apoptotic processes in brain. Overall effects of oxime reactivators are discussed in this study. The pertinent adverse effects and strong modulation of disparate parameters of oxime reactivators, HI-6 and obidoxime are not well understood in antidotal treatment. We found strong impact of oxime reactivators on redox homeostasis and apoptotic processes. The Ministry of Defence of the Czech Republic is gratefully acknowledged for the project no. OVUOFVZ200802 and The Ministry of Education, Youth and Sports is gratefully acknowledged for project Adjuvans. doi:10.1016/j.toxlet.2011.05.932
P2309 Oxidative stress and Na+ -glucose cotransporters Sglt1 and Sglt2 in kidneys of ochratoxin A-treated rats M. Peraica 1,∗ , D. Flajs 1 , M. Mladinic 2 , D. Zeljezic 2 , D. Balen Eror 3 , H. Koepsell 4 , I. Sabolic´ 3 1
Unit of Toxicology, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 2 Mutagenesis Unit, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 3 Unit of Molecular Toxicology, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 4 Unit of Anatomy and Cell Biology, University of Wurzburg, Wurzburg, Germany
OTA is a mould metabolite which contaminates food and feed, and causes nephrotoxicity by the poorly understood mechanism. In this study, male Wistar rats (N = 6) were gavaged with 0, 250 and 500 g OTA/kg b.m./day for 7 days. The 24-h urine was collected in metabolic cages the day before, and after 3 and 6 days of treatment. In animals sacrificed 24 h after the last treatment, blood plasma and kidneys were collected for biochemical and immunochemical analyses. Concentration of glutathione in plasma (mol L−1 ) decreased significantly in animals treated with 250 g (7.28 ± 1.95) and 500 g OTA (8.50 ± 1.30) (controls: 11.00 ± 2.05; mean ± SD; p < 0.05), but remained unchanged in kidney. Plasma malondialdehyde concentration increased with the OTA dose (0.52 ± 0.08, 0.79 ± 0.06 and 0.93 ± 0.11 mol L−1 , respectively). In the hOGG1 comet test, the tail intensity significantly increased in the kidney homogenate of the OTA-treated rats (2.76 ± 10.18% and 3.74 ± 9.88%, respectively) as compared to controls (1.54 ± 1.90%), while the comet tail length was significantly longer only at 500 g OTA (13.12 ± 6.41 m) than in controls (11.75 ± 5.06 m). A transient glucosuria was observed after 3 days of OTA treatment. Western blotting of isolated membranes and immunostaining of the p-formaldehyde fixed tissue cryosections revealed higher expression of Sglt1 and focal damage of proximal tubules (PT) at both OTA doses, and lower expression of Sglt2 at higher dose. The data indicate that OTA treatment in rats induces oxidative stress, and causes transient glucosuria by affecting the expression of glucose transporters and the structure of PT. doi:10.1016/j.toxlet.2011.05.933
S275
P2310 Role of cytoskeleton, cell proliferation and tight junction in wound healing: New therapeutic strategy K. Ghazi 1 , M. Dutot 1 , O. Laprévote 2 , J. Warnet 2 , P. Rat 3,∗ 1
Laboratoire de Chimie et Toxicologie Analytique et Cellulaire, Faculté de Pharmacie, Université Paris DESCARTES, Paris, France, 2 Laboratoire de Chimie et Toxicologie Analytique et Cellulaire, EA 4463, Faculté des Sciences Pharmaceutiques et biologiques, Paris, France, 3 Laboratoire de Toxicologie Analytique et Cellulaire (c-tac), Faculté de pharmacie, Université Paris Descartes, Paris, France Introduction: Skin is a tissue which envelops the body as a protective outer layer. Wound healing stages include inflammation, cell proliferation and migration. Calophyllum Inophyllum oil is tropical nut oil used in traditional medicine. It has wound healing and anti-inflammatory effects. Hyaluronan is the major component of the extracellular matrix, it binds to cells via CD44; this interaction induces cell migration and proliferation. P2X7 receptor is a purinergic receptor witch is activated by extracellular ATP; its basal activation induces cell growth. Calophyllum inophyllum and hyaluronan are both known to modulate P2X7 receptor. The aim of this study was to investigate wound healing effect through P2X7 receptor modulation of this tropical nut oil and hyaluronan on human keratinocytes cells (HaCat). Methods: Tropical nut oil was incubated for 15 min with HaCat cells after or before scratch test. After scratch test cells were washed and incubated with culture cell medium at 2.5% of FCS. Hyaluronan diluted in culture cell medium at 2.5% of FCS was incubated with HaCat after scratch test for 24 h. To determine if P2X7 plays a role in wound healing, we incubated HaCat cells for 20 minutes with Brillant Blue G (BBG) an inhibitor of P2X7 receptor activation before wound healing protocol. Cell proliferation was analysed and the distance between the edges of wound was measured at J0 and J1 (after 24 h). Tight junction expression (ZO-1) was analysed using flow cytometry assay. Results: Hyaluronan at 0.2% induced cell proliferation, wound healing and increase in ZO-1 expression compared to control. We also observed that tropical nut oil induced significant wound healing both when it is preincubated and post-incubated. P2X7 seems to have a critical role in cell growth. Conclusion: Wound effect of hyaluronan could involve cytoskeleton rearrangement. Tropical nut oil could be used before surgical intervention to stimulate wound healing, and after to improve scar aesthetic. doi:10.1016/j.toxlet.2011.05.934
P2311 Protective effect of thymoquinone against liver toxicity induced by aflatoxin B1 in mice O. Sabzevari 1,∗ , A. Nili-Ahmadabadi 1 , F. Tavakoli 1 , H. Rahimi 1 , H. Paydar 1 , G. Hasanzadeh 2 1 Toxicology and Pharmacology, Tehran University of Medical Sciences, Tehran, Iran, 2 Department of Histopathology, Tehran University of Medical Sciences, Tehran, Iran
Purpose: Thymoquinone (TQ) is one of the active components of Nigella sativa. The plant has been used in herbal medicine for the treatment of many diseases including liver complications. The present study aimed to investigate the protective effects of TQ on aflatoxin B1 (AFB1 ) induced liver toxicity of male mice. Methods: Animals were divided into six groups and treated intraperitoneally. Groups 1 (blank) and 2 (positive control) received vehicle, Group
caspase 3 and glutathione S-transferase were assessed using standard protocols. Results: We found significant depletion of liver and brain low molecular weight antioxidants. On the other hand, the depletion was partially recovered by an increase in the GSH level. Obidoxime was implicated in alteration of apoptotic processes in brain. Overall effects of oxime reactivators are discussed in this study. The pertinent adverse effects and strong modulation of disparate parameters of oxime reactivators, HI-6 and obidoxime are not well understood in antidotal treatment. We found strong impact of oxime reactivators on redox homeostasis and apoptotic processes. The Ministry of Defence of the Czech Republic is gratefully acknowledged for the project no. OVUOFVZ200802 and The Ministry of Education, Youth and Sports is gratefully acknowledged for project Adjuvans. doi:10.1016/j.toxlet.2011.05.932
P2309 Oxidative stress and Na+ -glucose cotransporters Sglt1 and Sglt2 in kidneys of ochratoxin A-treated rats M. Peraica 1,∗ , D. Flajs 1 , M. Mladinic 2 , D. Zeljezic 2 , D. Balen Eror 3 , H. Koepsell 4 , I. Sabolic´ 3 1
Unit of Toxicology, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 2 Mutagenesis Unit, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 3 Unit of Molecular Toxicology, Institute for Medical Research and Occupational Health, Zagreb, Croatia, 4 Unit of Anatomy and Cell Biology, University of Wurzburg, Wurzburg, Germany
OTA is a mould metabolite which contaminates food and feed, and causes nephrotoxicity by the poorly understood mechanism. In this study, male Wistar rats (N = 6) were gavaged with 0, 250 and 500 g OTA/kg b.m./day for 7 days. The 24-h urine was collected in metabolic cages the day before, and after 3 and 6 days of treatment. In animals sacrificed 24 h after the last treatment, blood plasma and kidneys were collected for biochemical and immunochemical analyses. Concentration of glutathione in plasma (mol L−1 ) decreased significantly in animals treated with 250 g (7.28 ± 1.95) and 500 g OTA (8.50 ± 1.30) (controls: 11.00 ± 2.05; mean ± SD; p < 0.05), but remained unchanged in kidney. Plasma malondialdehyde concentration increased with the OTA dose (0.52 ± 0.08, 0.79 ± 0.06 and 0.93 ± 0.11 mol L−1 , respectively). In the hOGG1 comet test, the tail intensity significantly increased in the kidney homogenate of the OTA-treated rats (2.76 ± 10.18% and 3.74 ± 9.88%, respectively) as compared to controls (1.54 ± 1.90%), while the comet tail length was significantly longer only at 500 g OTA (13.12 ± 6.41 m) than in controls (11.75 ± 5.06 m). A transient glucosuria was observed after 3 days of OTA treatment. Western blotting of isolated membranes and immunostaining of the p-formaldehyde fixed tissue cryosections revealed higher expression of Sglt1 and focal damage of proximal tubules (PT) at both OTA doses, and lower expression of Sglt2 at higher dose. The data indicate that OTA treatment in rats induces oxidative stress, and causes transient glucosuria by affecting the expression of glucose transporters and the structure of PT. doi:10.1016/j.toxlet.2011.05.933
S275
P2310 Role of cytoskeleton, cell proliferation and tight junction in wound healing: New therapeutic strategy K. Ghazi 1 , M. Dutot 1 , O. Laprévote 2 , J. Warnet 2 , P. Rat 3,∗ 1
Laboratoire de Chimie et Toxicologie Analytique et Cellulaire, Faculté de Pharmacie, Université Paris DESCARTES, Paris, France, 2 Laboratoire de Chimie et Toxicologie Analytique et Cellulaire, EA 4463, Faculté des Sciences Pharmaceutiques et biologiques, Paris, France, 3 Laboratoire de Toxicologie Analytique et Cellulaire (c-tac), Faculté de pharmacie, Université Paris Descartes, Paris, France Introduction: Skin is a tissue which envelops the body as a protective outer layer. Wound healing stages include inflammation, cell proliferation and migration. Calophyllum Inophyllum oil is tropical nut oil used in traditional medicine. It has wound healing and anti-inflammatory effects. Hyaluronan is the major component of the extracellular matrix, it binds to cells via CD44; this interaction induces cell migration and proliferation. P2X7 receptor is a purinergic receptor witch is activated by extracellular ATP; its basal activation induces cell growth. Calophyllum inophyllum and hyaluronan are both known to modulate P2X7 receptor. The aim of this study was to investigate wound healing effect through P2X7 receptor modulation of this tropical nut oil and hyaluronan on human keratinocytes cells (HaCat). Methods: Tropical nut oil was incubated for 15 min with HaCat cells after or before scratch test. After scratch test cells were washed and incubated with culture cell medium at 2.5% of FCS. Hyaluronan diluted in culture cell medium at 2.5% of FCS was incubated with HaCat after scratch test for 24 h. To determine if P2X7 plays a role in wound healing, we incubated HaCat cells for 20 minutes with Brillant Blue G (BBG) an inhibitor of P2X7 receptor activation before wound healing protocol. Cell proliferation was analysed and the distance between the edges of wound was measured at J0 and J1 (after 24 h). Tight junction expression (ZO-1) was analysed using flow cytometry assay. Results: Hyaluronan at 0.2% induced cell proliferation, wound healing and increase in ZO-1 expression compared to control. We also observed that tropical nut oil induced significant wound healing both when it is preincubated and post-incubated. P2X7 seems to have a critical role in cell growth. Conclusion: Wound effect of hyaluronan could involve cytoskeleton rearrangement. Tropical nut oil could be used before surgical intervention to stimulate wound healing, and after to improve scar aesthetic. doi:10.1016/j.toxlet.2011.05.934
P2311 Protective effect of thymoquinone against liver toxicity induced by aflatoxin B1 in mice O. Sabzevari 1,∗ , A. Nili-Ahmadabadi 1 , F. Tavakoli 1 , H. Rahimi 1 , H. Paydar 1 , G. Hasanzadeh 2 1 Toxicology and Pharmacology, Tehran University of Medical Sciences, Tehran, Iran, 2 Department of Histopathology, Tehran University of Medical Sciences, Tehran, Iran
Purpose: Thymoquinone (TQ) is one of the active components of Nigella sativa. The plant has been used in herbal medicine for the treatment of many diseases including liver complications. The present study aimed to investigate the protective effects of TQ on aflatoxin B1 (AFB1 ) induced liver toxicity of male mice. Methods: Animals were divided into six groups and treated intraperitoneally. Groups 1 (blank) and 2 (positive control) received vehicle, Group