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Use of ex-vivo testis culture to explore mechanisms of seminiferous tubular atrophy upon PIM inhibition
S164
Abstracts / Toxicology Letters 280S (2017) S162–S165
tion seeking for reliable markers of hypoxic insult. Neither maternal mortality nor pre-term birth was observed. Fetal growth reduction was seen in group of 12 h eGIH only. eGIH (8 h and 12 h) at 15th16th days of gestation DG resulted in downregulation of the levels of heme oxygenase-1, a ubiquitous inducible antioxidant stress protein, and upregulation of the levels of IgG in the amniotic fluid, as assessed by western blot analysis. These data suggest that eGIH can affect the immune and redox homeostasis in the developing fetus lasting until delivery. Histological examination of placental tissue after eGIH revealed perivascular edema in both intervals and arterial wall thickening after 12 h of eGIH. In conclusion this model was able to induce persistent changes in crucial components of development without producing visible harmful effects on mother or fetus. Acknowledgment: This work was supported by the grant VEGA 2/0129/15 and 2/0166/16. http://dx.doi.org/10.1016/j.toxlet.2017.07.458 P-04-10-07 Assessment of oxidative stress inducing effects of perfluorooctanoic acid in mice testes and possible preventive effect of taurine Burcu Ünlü Endirlik 1,2 , Ays¸e Eken 1 , Figen Öztürk 3 , Aylin Gürbay 2 1 Department of Toxicology, Faculty of Pharmacy, Erciyes University, Kayseri, Turkey 2 Department of Toxicology, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey 3 Department of Pathology, Faculty of Medicine, Erciyes University, Kayseri, Turkey
Perfluorooctanoic acid (PFOA) is a persistent compound used in several industrial processes, and commonly detected in humans, wildlife and in the environment. Disruptive adverse outcomes of PFOA on reproductive and developmental systems have been reported besides its other toxic effects. The purpose of the present study was to investigate oxidative stress inducing effects of PFOA in testes of Balb/c mice following administration of the compound in two different concentrations for 10 days. Results of this study showed that PFOA (15 or 30 mg/kg/day, ig, for 10 days) altered oxidative stress parameters in testes of mice, as evident by slight and significant increases in malondialdehyde (in both dose groups), and total glutathione levels (in 15 mg/kg dose group), respectively, and additionally, significant reductions on the activities of antioxidant enzymes. It was also found that PFOA (15 mg/kg) caused a significant decrease in testes weights of mice. Histopathological examinations showed that PFOA did not induce any changes in these organs. Taurine pretreatment (100 mg/kg/d for 5 days) preceding the administration of high dose PFOA (for 10 days) caused significant increases in the activities of catalase and GPx. These results indicate that PFOA treatment introduces an oxidative stress in testes of mice. Further studies are required in order to better understand the causes of the organ weight reducing and oxidative stress inducing effects of PFOA in this model system. Acknowledgment: This study was supported by Hacettepe University Research Foundation (013 D05 301 001). http://dx.doi.org/10.1016/j.toxlet.2017.07.459
P-04-10-08 Use of ex-vivo testis culture to explore mechanisms of seminiferous tubular atrophy upon PIM inhibition Marie H. Larsson 1 , Ulf Andersson 2 , Jayne Harris 3 , Jorrit J. Hornberg 1 , Terri Mitchard 3 1
Discovery Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Gothenburg, Gothenburg, Sweden 2 Regulatory Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Gothenburg, Gothenburg, Sweden 3 Regulatory Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Cambridge, Cambridge, United Kingdom In order to ensure design and selection of innovative drug candidates with the Right Safety profile, we employ a pro-active Discovery Safety strategy. This encompasses early identification of potential safety concerns and subsequent mitigation by influencing chemical design. Seminiferous tubular atrophy was found in rat after oral dosing for 8 days with a PIM inhibitor (Proto-oncogene serine/threonine-protein kinases). As PIMs have been considered as drug targets for various disease indications and are expressed in testis, a 3D ex-vivo seminiferous tubule model of spermatogenesis were applied to evaluate the translation of the in vivo findings to an ex-vivo model and the potential mechanisms of the seminiferous tubular atrophy. Three PIM inhibitors, with different potencies versus PIM1, 2 and 3 were chosen to identify relative contribution of the PIM members and thereby inform the medicinal chemistry program how to steer away from testicular toxicity by optimizing PIM selectivity. After 7 days treatment, a dose-dependent decrease of spermatogonia (60–80% vs. control at top concentration) and a dose-independent increase of young pachytene spermatocytes (120–160% vs. control) was observed. Pan-PIM inhibition were found to cause a larger decrease in secondary spermatocytes (60% vs. 80% of control) and round spermatids (50% vs. 90% of control) compared to PIM1 inhibition alone. These data suggest that in vivo and ex-vivo findings are consistent and that PIM inhibitors have negative effects on the spermatogenic process, with pan-PIM inhibition being more deleterious. http://dx.doi.org/10.1016/j.toxlet.2017.07.460 P-04-10-09 Treatment of human placenta choriocarcinoma cells with two components of cigarette smoke induced growth and epithelial mesenchymal transition via induction of an antioxidant effect Hae-Miru Lee, Kyung-Chul Choi Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea In this study, the effects of formaldehyde (FA) and benzene (Bz) were examined on cell proliferation and epithelial mesenchymal transition (EMT) of JEG-3 human choriocarcinoma cells to confirm relationship between CS components and placenta carcinoma. Upon MTT assay, FA (10−8 M to 10−5 M) and Bz (10−11 M to 10−5 M) increased JEG-3 cell proliferation. Western blot assay revealed that the protein expression of cyclin D1 & E1 increased, while the levels of p21 & p27 were reduced following treatment. In addition, the expression of the epithelial marker, E-cadherin, was significantly
Abstracts / Toxicology Letters 280S (2017) S162–S165
tion seeking for reliable markers of hypoxic insult. Neither maternal mortality nor pre-term birth was observed. Fetal growth reduction was seen in group of 12 h eGIH only. eGIH (8 h and 12 h) at 15th16th days of gestation DG resulted in downregulation of the levels of heme oxygenase-1, a ubiquitous inducible antioxidant stress protein, and upregulation of the levels of IgG in the amniotic fluid, as assessed by western blot analysis. These data suggest that eGIH can affect the immune and redox homeostasis in the developing fetus lasting until delivery. Histological examination of placental tissue after eGIH revealed perivascular edema in both intervals and arterial wall thickening after 12 h of eGIH. In conclusion this model was able to induce persistent changes in crucial components of development without producing visible harmful effects on mother or fetus. Acknowledgment: This work was supported by the grant VEGA 2/0129/15 and 2/0166/16. http://dx.doi.org/10.1016/j.toxlet.2017.07.458 P-04-10-07 Assessment of oxidative stress inducing effects of perfluorooctanoic acid in mice testes and possible preventive effect of taurine Burcu Ünlü Endirlik 1,2 , Ays¸e Eken 1 , Figen Öztürk 3 , Aylin Gürbay 2 1 Department of Toxicology, Faculty of Pharmacy, Erciyes University, Kayseri, Turkey 2 Department of Toxicology, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey 3 Department of Pathology, Faculty of Medicine, Erciyes University, Kayseri, Turkey
Perfluorooctanoic acid (PFOA) is a persistent compound used in several industrial processes, and commonly detected in humans, wildlife and in the environment. Disruptive adverse outcomes of PFOA on reproductive and developmental systems have been reported besides its other toxic effects. The purpose of the present study was to investigate oxidative stress inducing effects of PFOA in testes of Balb/c mice following administration of the compound in two different concentrations for 10 days. Results of this study showed that PFOA (15 or 30 mg/kg/day, ig, for 10 days) altered oxidative stress parameters in testes of mice, as evident by slight and significant increases in malondialdehyde (in both dose groups), and total glutathione levels (in 15 mg/kg dose group), respectively, and additionally, significant reductions on the activities of antioxidant enzymes. It was also found that PFOA (15 mg/kg) caused a significant decrease in testes weights of mice. Histopathological examinations showed that PFOA did not induce any changes in these organs. Taurine pretreatment (100 mg/kg/d for 5 days) preceding the administration of high dose PFOA (for 10 days) caused significant increases in the activities of catalase and GPx. These results indicate that PFOA treatment introduces an oxidative stress in testes of mice. Further studies are required in order to better understand the causes of the organ weight reducing and oxidative stress inducing effects of PFOA in this model system. Acknowledgment: This study was supported by Hacettepe University Research Foundation (013 D05 301 001). http://dx.doi.org/10.1016/j.toxlet.2017.07.459
P-04-10-08 Use of ex-vivo testis culture to explore mechanisms of seminiferous tubular atrophy upon PIM inhibition Marie H. Larsson 1 , Ulf Andersson 2 , Jayne Harris 3 , Jorrit J. Hornberg 1 , Terri Mitchard 3 1
Discovery Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Gothenburg, Gothenburg, Sweden 2 Regulatory Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Gothenburg, Gothenburg, Sweden 3 Regulatory Safety, Drug Safety and Metabolism, Innovative Medicines and Early Development, AstraZeneca R&D Cambridge, Cambridge, United Kingdom In order to ensure design and selection of innovative drug candidates with the Right Safety profile, we employ a pro-active Discovery Safety strategy. This encompasses early identification of potential safety concerns and subsequent mitigation by influencing chemical design. Seminiferous tubular atrophy was found in rat after oral dosing for 8 days with a PIM inhibitor (Proto-oncogene serine/threonine-protein kinases). As PIMs have been considered as drug targets for various disease indications and are expressed in testis, a 3D ex-vivo seminiferous tubule model of spermatogenesis were applied to evaluate the translation of the in vivo findings to an ex-vivo model and the potential mechanisms of the seminiferous tubular atrophy. Three PIM inhibitors, with different potencies versus PIM1, 2 and 3 were chosen to identify relative contribution of the PIM members and thereby inform the medicinal chemistry program how to steer away from testicular toxicity by optimizing PIM selectivity. After 7 days treatment, a dose-dependent decrease of spermatogonia (60–80% vs. control at top concentration) and a dose-independent increase of young pachytene spermatocytes (120–160% vs. control) was observed. Pan-PIM inhibition were found to cause a larger decrease in secondary spermatocytes (60% vs. 80% of control) and round spermatids (50% vs. 90% of control) compared to PIM1 inhibition alone. These data suggest that in vivo and ex-vivo findings are consistent and that PIM inhibitors have negative effects on the spermatogenic process, with pan-PIM inhibition being more deleterious. http://dx.doi.org/10.1016/j.toxlet.2017.07.460 P-04-10-09 Treatment of human placenta choriocarcinoma cells with two components of cigarette smoke induced growth and epithelial mesenchymal transition via induction of an antioxidant effect Hae-Miru Lee, Kyung-Chul Choi Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea In this study, the effects of formaldehyde (FA) and benzene (Bz) were examined on cell proliferation and epithelial mesenchymal transition (EMT) of JEG-3 human choriocarcinoma cells to confirm relationship between CS components and placenta carcinoma. Upon MTT assay, FA (10−8 M to 10−5 M) and Bz (10−11 M to 10−5 M) increased JEG-3 cell proliferation. Western blot assay revealed that the protein expression of cyclin D1 & E1 increased, while the levels of p21 & p27 were reduced following treatment. In addition, the expression of the epithelial marker, E-cadherin, was significantly