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Fumonisin B1 affects “in vitro” intestinal epithelial cell barrier integrity and reduces claudin expression
Abstracts / Toxicology Letters 258S (2016) S62–S324
P16-073 Expression of H3K9ac, H3K9me2 mediated by signal pathway of NMDAR-ERK in the hippocampus of rats chronically exposed to aluminum Q. Niu ∗ , Z. Li, P. Kang, Q. Zhang, X. Lu, J. Nie, J. Song, L. Wang School of Public Health, Shanxi Medical University, Taiyuan, China To explore the impact of chronic aluminum exposure on learning and memory ability through histone modification by studying the changes of N-methyl-d-aspartic acid receptor (NMDAR), extracellular regulated protein kinases (Phospho-ERK) H3K9ac, H3K9me2, HP1, and BDNF in rat hippocampus. Methods: 24 healthy SPF grade SD male rats were randomly divided into four groups by weight, including control group and low, medium, high dose aluminum exposed group, 6 rats in each group. The Al-exposed rats drank water containing different doses of aluminum chloride (AlCl3 ) (2, 12, and 72 mg/kg Al3+ ) for 120 days, expression of NMDAR, P-ERK, H3K9ac, H3K9me2, HP1, and BNDNF was detected with Western blot. Results: The results of Western-blot test demonstrated that the expression of NMDAR in the Al-exposed groups were significantly lower than those of the control group (P < 0.05), and P-ERK, H3K9ac, and BDNF in the Al-exposed groups were significantly lower than those of the control group too (P < 0.01), while the expressions of H3K9me2 and HP1in the Al-exposed groups were both significantly higher than those in the control group (P < 0.05). Conclusion: Chronic aluminum exposure may change the histone modification via inhibiting the signal pathway of NMDAR-ERK, and therefore impair the ability of learning and memory in rats. http://dx.doi.org/10.1016/j.toxlet.2016.06.1912 P16-074 Fumonisin B1 affects “in vitro” intestinal epithelial cell barrier integrity and reduces claudin expression ∗ , M.A. Martínez, I. Ares, E. Ramos, V. ˜ M.R. Martínez Larranaga Castellano, M. Martínez, A. Anadón, A. Romero
Department of Toxicology and Pharmacology, Faculty of Veterinary Medicine, Universidad Complutense de Madrid, 28040 Madrid, Spain Although the literature about mycotoxins is rich in reports investigating cellular mechanisms, cellular toxicity, associated pathology and animal performance, studies on the effect of these compounds on the gastrointestinal tract is limited. The intestinal tract represents the first barrier to ingested chemicals or food contaminants. The gut barrier is formed to a large extent by tight junctions, the most important components of the intercellular junctional complexes regulating cell adhesion and selective permeability in the gut. Because of transmembrane proteins claudins and occludin determine adhesion and permeability characteristics of the epithelium in terms of specificity and tightness, the aim of the present study was to evaluate the expression of claudin-3, claudin4 and occludin in Caco-2 cells exposed to Fumonisin 1 (FB1). Our results showed that FB1 treatment strongly reduced the expression of claudin-3, claudin-4 and occludin. The decrease of mRNA expression of claudin-3 reached values of 29%, 36%, 38% and 60% for cells treated with 1, 3, 10 and 30 M FB1, respectively. The decrease of mRNA expression of claudin-4 reached values of 40%, 58%, 60% and 80% for cells treated with 1, 3, 10 and 30 M FB1, respectively. The decrease of mRNA expression of occludin reached values of 70%,
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80%, 80% and 92% for cells treated with 1, 3, 10 and 30 M FB1, respectively. These observations may help to explain, at a molecular level, some of the “in vivo” effects of these mycotoxins on the intestine. Acknowledgements: Work supported by Project S2013/ABI2728 (Programa ALIBIRD-Comunidad de Madrid). http://dx.doi.org/10.1016/j.toxlet.2016.06.1913 P16-075 Cytotoxicity of deoxynivalenol and cytoprotection of antioxidants in Caco-2 cells M. Iula, C. Fernández Blanco, A. Juan García, G. Font, M. Ruiz ∗ Laboratory of Toxicology, Faculty of Pharmacy, University of Valencia, Valencia, Spain Deoxynivalenol (DON) is a secondary metabolite of Fusarium fungi producing cell injury. When antioxidant agents (polyphenols and vitamins), commonly present in vegetables, are simultaneously ingested with DON, cytotoxic effects can be interfered. Aims of this work were to determine mechanisms of DON-inducing cell damage and cytoprotection effect of antioxidants in Caco-2 cells. Cytotoxic effect of DON was determined by MTT assay. Reactive oxygen species (ROS) production and lipid peroxidation (LPO) were determined using the fluorescein and TBARS assays, respectively. To determine cytoprotection of antioxidant, Caco-2 cells previously exposed to different concentrations of quercetin, pterostilbene, vitamin C and vitamin E (5, 25 and 50 M) were treated with DON. IC50 values obtained were 9.5 (48 h) and 7 (72 h) M, while no IC50 value was obtained after 24 h. Oxidative stress and cellular antioxidant protection were assayed at 0.5 and 1 M DON. LPO increased by 33% after 1 M DON. ROS generation increased in time dependent manner. The highest increases obtained were 30% (0.5 M DON) and 40% (1 M DON) at 15 min exposure. Cell viability increased at 25 and 50 M vitamin C and quercetin, respectively compared to cells exposed to DON; increases were from 20% to 25% and from 12% to 28% for vitamin C and quercetin, respectively. Cytotoxicity of DON was prevented by vitamin C and quercetin. Acknowledgments: This work was supported by Spanish Ministry of Economy and Competitiveness (AGL2013-43194-P). http://dx.doi.org/10.1016/j.toxlet.2016.06.1914 P16-076 Effects of curcumin on cisplatin cytotoxicity in HeLa cells S. Aydin 1,∗ , M. Becit 1 , A. Basaran 2 , N. Basaran 1 1
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Hacettepe University, 06100 Ankara, Turkey 2 Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, 06100 Ankara, Turkey Phenolic compounds, because of their antioxidant properties, have an important role in the prevention of aging, cancer, and several degenerative and genetic diseases. Curcumin, a phenolic compound, obtained from Curcuma longa, has some beneficial health effects such as antioxidant, antiinflammatory, anticarcinogenic, and antimutagenic effects. The combination of some plant derived compounds, especially phenolic compounds, with antineoplastic drugs such as cisplatin has been suggested to increase the anticancer effects and also decrease their adverse effects in the chemotherapy. However, there are limited studies
P16-073 Expression of H3K9ac, H3K9me2 mediated by signal pathway of NMDAR-ERK in the hippocampus of rats chronically exposed to aluminum Q. Niu ∗ , Z. Li, P. Kang, Q. Zhang, X. Lu, J. Nie, J. Song, L. Wang School of Public Health, Shanxi Medical University, Taiyuan, China To explore the impact of chronic aluminum exposure on learning and memory ability through histone modification by studying the changes of N-methyl-d-aspartic acid receptor (NMDAR), extracellular regulated protein kinases (Phospho-ERK) H3K9ac, H3K9me2, HP1, and BDNF in rat hippocampus. Methods: 24 healthy SPF grade SD male rats were randomly divided into four groups by weight, including control group and low, medium, high dose aluminum exposed group, 6 rats in each group. The Al-exposed rats drank water containing different doses of aluminum chloride (AlCl3 ) (2, 12, and 72 mg/kg Al3+ ) for 120 days, expression of NMDAR, P-ERK, H3K9ac, H3K9me2, HP1, and BNDNF was detected with Western blot. Results: The results of Western-blot test demonstrated that the expression of NMDAR in the Al-exposed groups were significantly lower than those of the control group (P < 0.05), and P-ERK, H3K9ac, and BDNF in the Al-exposed groups were significantly lower than those of the control group too (P < 0.01), while the expressions of H3K9me2 and HP1in the Al-exposed groups were both significantly higher than those in the control group (P < 0.05). Conclusion: Chronic aluminum exposure may change the histone modification via inhibiting the signal pathway of NMDAR-ERK, and therefore impair the ability of learning and memory in rats. http://dx.doi.org/10.1016/j.toxlet.2016.06.1912 P16-074 Fumonisin B1 affects “in vitro” intestinal epithelial cell barrier integrity and reduces claudin expression ∗ , M.A. Martínez, I. Ares, E. Ramos, V. ˜ M.R. Martínez Larranaga Castellano, M. Martínez, A. Anadón, A. Romero
Department of Toxicology and Pharmacology, Faculty of Veterinary Medicine, Universidad Complutense de Madrid, 28040 Madrid, Spain Although the literature about mycotoxins is rich in reports investigating cellular mechanisms, cellular toxicity, associated pathology and animal performance, studies on the effect of these compounds on the gastrointestinal tract is limited. The intestinal tract represents the first barrier to ingested chemicals or food contaminants. The gut barrier is formed to a large extent by tight junctions, the most important components of the intercellular junctional complexes regulating cell adhesion and selective permeability in the gut. Because of transmembrane proteins claudins and occludin determine adhesion and permeability characteristics of the epithelium in terms of specificity and tightness, the aim of the present study was to evaluate the expression of claudin-3, claudin4 and occludin in Caco-2 cells exposed to Fumonisin 1 (FB1). Our results showed that FB1 treatment strongly reduced the expression of claudin-3, claudin-4 and occludin. The decrease of mRNA expression of claudin-3 reached values of 29%, 36%, 38% and 60% for cells treated with 1, 3, 10 and 30 M FB1, respectively. The decrease of mRNA expression of claudin-4 reached values of 40%, 58%, 60% and 80% for cells treated with 1, 3, 10 and 30 M FB1, respectively. The decrease of mRNA expression of occludin reached values of 70%,
S259
80%, 80% and 92% for cells treated with 1, 3, 10 and 30 M FB1, respectively. These observations may help to explain, at a molecular level, some of the “in vivo” effects of these mycotoxins on the intestine. Acknowledgements: Work supported by Project S2013/ABI2728 (Programa ALIBIRD-Comunidad de Madrid). http://dx.doi.org/10.1016/j.toxlet.2016.06.1913 P16-075 Cytotoxicity of deoxynivalenol and cytoprotection of antioxidants in Caco-2 cells M. Iula, C. Fernández Blanco, A. Juan García, G. Font, M. Ruiz ∗ Laboratory of Toxicology, Faculty of Pharmacy, University of Valencia, Valencia, Spain Deoxynivalenol (DON) is a secondary metabolite of Fusarium fungi producing cell injury. When antioxidant agents (polyphenols and vitamins), commonly present in vegetables, are simultaneously ingested with DON, cytotoxic effects can be interfered. Aims of this work were to determine mechanisms of DON-inducing cell damage and cytoprotection effect of antioxidants in Caco-2 cells. Cytotoxic effect of DON was determined by MTT assay. Reactive oxygen species (ROS) production and lipid peroxidation (LPO) were determined using the fluorescein and TBARS assays, respectively. To determine cytoprotection of antioxidant, Caco-2 cells previously exposed to different concentrations of quercetin, pterostilbene, vitamin C and vitamin E (5, 25 and 50 M) were treated with DON. IC50 values obtained were 9.5 (48 h) and 7 (72 h) M, while no IC50 value was obtained after 24 h. Oxidative stress and cellular antioxidant protection were assayed at 0.5 and 1 M DON. LPO increased by 33% after 1 M DON. ROS generation increased in time dependent manner. The highest increases obtained were 30% (0.5 M DON) and 40% (1 M DON) at 15 min exposure. Cell viability increased at 25 and 50 M vitamin C and quercetin, respectively compared to cells exposed to DON; increases were from 20% to 25% and from 12% to 28% for vitamin C and quercetin, respectively. Cytotoxicity of DON was prevented by vitamin C and quercetin. Acknowledgments: This work was supported by Spanish Ministry of Economy and Competitiveness (AGL2013-43194-P). http://dx.doi.org/10.1016/j.toxlet.2016.06.1914 P16-076 Effects of curcumin on cisplatin cytotoxicity in HeLa cells S. Aydin 1,∗ , M. Becit 1 , A. Basaran 2 , N. Basaran 1 1
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Hacettepe University, 06100 Ankara, Turkey 2 Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, 06100 Ankara, Turkey Phenolic compounds, because of their antioxidant properties, have an important role in the prevention of aging, cancer, and several degenerative and genetic diseases. Curcumin, a phenolic compound, obtained from Curcuma longa, has some beneficial health effects such as antioxidant, antiinflammatory, anticarcinogenic, and antimutagenic effects. The combination of some plant derived compounds, especially phenolic compounds, with antineoplastic drugs such as cisplatin has been suggested to increase the anticancer effects and also decrease their adverse effects in the chemotherapy. However, there are limited studies