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Cadmium exposure increases the susceptibility to testicular autoimmunity in mice
Abstracts for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20
thrombogenic reactions. Our results raise the possibility that sRAGE may be a novel biomarker of RPL.
Goshajinkigan completely recover the severe aspermatogenesis after busulfan treatment in mice
http://dx.doi.org/10.1016/j.jri.2014.09.025
N. Qu ∗ , Y. Hirayanagi, S. Hayashi, S. Hirai, N. Hatayama, M. Kuramasu, Y. Ogawa, M. Itoh
DR6 is epigenetically involved in the pathogenesis of endometriosis by creating the proliferative and anti-apoptotic characteristics Kai 1,∗ ,
Nasu 1,2 ,
Aoyagi 1 ,
Tsukamoto 3 ,
K. K. Y. Y. N. Hijiya 3 , M. Okamoto 1 , M. Moriyama 3 , H. Narahara 1 1
Department of Obstetrics and Gynecology, Japan Division of Obstetrics and Gynecology, Support System for Community Medicine, Japan 3 Department of Molecular Pathology, Faculty of Medicine; Oita University, Oita 879-5593, Japan 2
Objective: Accumulating evidence suggests that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. The goals of this study were to identify the roles of death receptor (DR) 6, the only TNFRSF whose expression is epigenetically repressed in endometriotic cyst stromal cells (ECSCs), in the pathogenesis of endometriosis. Methods: We identified that DR6 was upregulated by valpronic acid (VPA) treatment in ECSCs based on a mRNA microarray data. The expression of DR6 protein was confirmed ex vivo and in vitro. The effects of DR6 on the cell proliferation and apoptosis were investigated in normal endometrial stromal cells (NESCs). Results: DR6 protein expression was attenuated in ECSCs, and its expression was upregulated by VPA stimulation. Immunohistochemical stainings confirmed the decreased staining for DR6 protein in endometriotic tissues. VPA treatment resulted in an accumulation of acetylated histones H4 in the promoter region of the DR6 gene in ECSCs. DR6 knockdown directed the stimulation of cell proliferation and the resistance to apoptosis in NESCs. Conclusions: Our findings suggest that DR6, an epigenetically suppressed TNFRSF gene is involved in the pathogenesis of endometriosis by creating the proliferative and anti-apoptotic characteristics of endometriosis. http://dx.doi.org/10.1016/j.jri.2014.09.026
11
Department of Anatomy, Tokyo Medical University, Tokyo 160-8402, Japan Busulfan (Myleran, 1,4-butanediol methanesulfonate) is used as anticancer chemotherapeutic drugs in childhood and adult chronic myelogenous leukemia as well as an immunosuppressive agent before bone marrow transplantation. It is well known that the male infertility including spermatogenesis disturbance is one of side effect. There is little information about therapeutic drugs on male infertility after busulfan treatment. Some traditional Chinese medicines have been administered for improving spermatogenesis and have been extensively used to treat male infertility. In the present study, we gave goshajinkigan (TJ107; Tsumura Co., Ltd., Tokyo, Japan) to mice already having severe aspermatogenesis after busulfan treatment to determine whether or not the goshajinkigan can recover the aspermatogenesis. Male C57BL/6j mice were received a single intraperitoneal injection of busulfan at 4-week-old and after 60 days fed on the goshajinkigan-including diet or goshajinkiganfree normal diet for another 60 days. The results showed that after busulfan treatment, the progressively decreases in the weight of the testes (TW) and epididymal sperm count (ESC) in normal diet group from 60 days (TW: 0.041 ± 0.0083 g; ESC: 1.575 ± 0.3079 × 105 cells) to 120 days (TW: 0.017 ± 0.0019 g; ESC: 0.22 ± 0.0189 × 105 cells); on the other hand, in goshajinkigan-including diet group, the dramatic recovery of these variables (TW: 0.10 ± 0.0062 g; ESC: 21.68 ± 1.705 × 105 cells) at 120 days, which is similarity to the normal spermatogenesis (TW: 0.098 ± 0.0024 g; ESC:19.8 ± 2.46 × 105 cells). These results suggest that busulfan-induced aspermatogenesis was irreversible unless receiving any medication. However, the supplementation of goshajinkigan can completely recover the regeneration of the injured semniferous epithelium, suggesting that goshajinkigan have a therapeutic effect on busulfan-induced aspermatogenesis. http://dx.doi.org/10.1016/j.jri.2014.09.027 Cadmium exposure increases the susceptibility to testicular autoimmunity in mice Y. Ogawa ∗ , M. Kuramasu, N. Qu, S. Hirai, S. Hayashi, N. Hatayama, M. Itoh Department of Anatomy, Tokyo Medical University, Japan Cadmium, one of various environmental toxicants, is known to suppress systemic immunity and to injure the testicular capillary endothelia with resultant
12
Abstracts for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20
necrosis of testicular tissues in mice and rats treated with high doses. Recently, it also became evident that cadmium can affect the integrity of the blood–testis barrier (BTB), the endocrine function of Leydig cells, apoptosis of germ cells and systemic immunity, even on treatment with a low dose that does not induce spermatogenic disturbance. Experimental autoimmune orchitis (EAO), i.e., an organ-specific autoimmunity of the testis, can be induced by repeated immunization with testicular antigens, and its pathology is characterized by lymphocytic inflammation and spermatogenic disturbance. In the present study, we investigated the morphological and functional changes of testes in mice treated with a low dose of cadmium chloride (CdCl2 ) and also examined its toxicity as to susceptibility to EAO. The results showed that exposure to 3 mg CdCl2/ kg body weight did not affect the spermatogenic state. However, the BTB at the tubuli recti and the rete testis, but not the seminiferous tubules, was slightly weakened, and intra-testicular mRNA expression of IL-6, TNF-␣ and IL-1 was significantly increased by the CdCl2 treatment. Furthermore, immunization with testicular antigens after the CdCl2 exposure significantly augmented the EAO severity. Therefore, exposure to a low dose of CdCl2 induces no significant disturbance of spermatogenesis, however, it does change the immunological microcircumstances in the testis, resulting in increased susceptibility to testicular autoimmunity. http://dx.doi.org/10.1016/j.jri.2014.09.028 Prevention of preterm labor by omega-3 polyunsaturated fatty acids and resolvin E3 E. Inoue ∗ , T. Nagamatsu, K. Kawana, T. Yamashita, Y. Osuga, T. Fujii Department of Obstetrics and Gynecology, Faculty of medicine, The University of Tokyo, Japan Introduction: Metabolites of omega-3 polyunsaturated fatty acids (PUFAs) have anti-inflammatory property, opposing inflammatory actions by omega-6-derived eicosanoids. Recently resolvin E3 is identified as a novel anti-inflammatory substance produced in the metabolic pathway of omega-3 PUFAs. Objective: This study aimed to investigate the possible role of omega 3 PUFAs in the regulation of intrauterine inflammation. We evaluated the relevance of omega-3 PUFAs and its metabolite, resolvin E3 to the pathology of preterm birth using a mouse model. Methods: Preterm birth was induced in mice by administering lipopolysaccharide (LPS) transcervically at day
15 pc. We examined the effect of resolvin E3 on the same model. Resolvin E3 was injected twice at the same time of LPS administration and at 6 h later to wild-type mice. Results: Preterm birth occurred in 87.5% in control mice and 56.3% in the mice after resolvin E3 injection (P = 0.03). The rate of fetal number left in the uterus was 34.9% in control mice and 65.3% in the mice with resolvin E3 injection (P < 0.01). Conclusions: We confirmed that omega-3 PUFAs suppress preterm birth induced by LPS. Our findings suggest that omega-3 PUFAs contribute to the prevention of inflammatory process caused by omega-6 PUFAs-derived eicosanoids induced by LPS injection. Anti-inflammatory metabolites of omega-3 PUFAs can be candidates for a new therapeutic medicine for preterm birth. http://dx.doi.org/10.1016/j.jri.2014.09.029 Interleukin-1 increases expression of tryptophan 2,3-dioxygenase and stimulates tryptophan metabolism in ectopic endomtetrial stromal cells Y. Urata ∗ , Y. Osuga, K. Koga, Y. Hirota, T. Hirata, M. Harada, M. Takamura, G. Izumi, T. Fujii The University of Tokyo, Department of Obstetrics and Gynecology, Japan Objective: Tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO) are enzymes that catalyze the first and rate-limiting step of tryptophan degradation (i.e. from tryptophan to kynurenine) and thereby regulate tryptophan levels. Decrease in tryptophan levels inhibits T cell function and contributes to immune tolerance. While IFN␥ increases IDO expression, a molecule that regulates TDO expression remains to be elucidated. We evaluated the effect of IL-1, a typical endometriosis-associated cytokine, on TDO expression in ectopic endometrial stromal cells (ESCs). Methods and results: ESCs were isolated from ovarian endometriomas and treated with IL-1. IL-1 (1 ng/ml) increased TDO mRNA expression to 6.3-fold of control but had no effect on IDO mRNA expression in ESCs. IL-1 increased TDO protein to 1.3-fold in ESCs and kynurenine production to 1.9-fold in the conditioned medium. TDO siRNA suppressed the increase in IL-1-induced kynurenine, IL-6 and IL-8 production. Conclusions: IL-1 increased TDO expression and stimulated tryptophan metabolism in ESCs. The IL-1-induced increase in TDO expression may enhance immune tolerance to promote endometriosis. http://dx.doi.org/10.1016/j.jri.2014.09.030
thrombogenic reactions. Our results raise the possibility that sRAGE may be a novel biomarker of RPL.
Goshajinkigan completely recover the severe aspermatogenesis after busulfan treatment in mice
http://dx.doi.org/10.1016/j.jri.2014.09.025
N. Qu ∗ , Y. Hirayanagi, S. Hayashi, S. Hirai, N. Hatayama, M. Kuramasu, Y. Ogawa, M. Itoh
DR6 is epigenetically involved in the pathogenesis of endometriosis by creating the proliferative and anti-apoptotic characteristics Kai 1,∗ ,
Nasu 1,2 ,
Aoyagi 1 ,
Tsukamoto 3 ,
K. K. Y. Y. N. Hijiya 3 , M. Okamoto 1 , M. Moriyama 3 , H. Narahara 1 1
Department of Obstetrics and Gynecology, Japan Division of Obstetrics and Gynecology, Support System for Community Medicine, Japan 3 Department of Molecular Pathology, Faculty of Medicine; Oita University, Oita 879-5593, Japan 2
Objective: Accumulating evidence suggests that various epigenetic aberrations play definite roles in the pathogenesis of endometriosis. The goals of this study were to identify the roles of death receptor (DR) 6, the only TNFRSF whose expression is epigenetically repressed in endometriotic cyst stromal cells (ECSCs), in the pathogenesis of endometriosis. Methods: We identified that DR6 was upregulated by valpronic acid (VPA) treatment in ECSCs based on a mRNA microarray data. The expression of DR6 protein was confirmed ex vivo and in vitro. The effects of DR6 on the cell proliferation and apoptosis were investigated in normal endometrial stromal cells (NESCs). Results: DR6 protein expression was attenuated in ECSCs, and its expression was upregulated by VPA stimulation. Immunohistochemical stainings confirmed the decreased staining for DR6 protein in endometriotic tissues. VPA treatment resulted in an accumulation of acetylated histones H4 in the promoter region of the DR6 gene in ECSCs. DR6 knockdown directed the stimulation of cell proliferation and the resistance to apoptosis in NESCs. Conclusions: Our findings suggest that DR6, an epigenetically suppressed TNFRSF gene is involved in the pathogenesis of endometriosis by creating the proliferative and anti-apoptotic characteristics of endometriosis. http://dx.doi.org/10.1016/j.jri.2014.09.026
11
Department of Anatomy, Tokyo Medical University, Tokyo 160-8402, Japan Busulfan (Myleran, 1,4-butanediol methanesulfonate) is used as anticancer chemotherapeutic drugs in childhood and adult chronic myelogenous leukemia as well as an immunosuppressive agent before bone marrow transplantation. It is well known that the male infertility including spermatogenesis disturbance is one of side effect. There is little information about therapeutic drugs on male infertility after busulfan treatment. Some traditional Chinese medicines have been administered for improving spermatogenesis and have been extensively used to treat male infertility. In the present study, we gave goshajinkigan (TJ107; Tsumura Co., Ltd., Tokyo, Japan) to mice already having severe aspermatogenesis after busulfan treatment to determine whether or not the goshajinkigan can recover the aspermatogenesis. Male C57BL/6j mice were received a single intraperitoneal injection of busulfan at 4-week-old and after 60 days fed on the goshajinkigan-including diet or goshajinkiganfree normal diet for another 60 days. The results showed that after busulfan treatment, the progressively decreases in the weight of the testes (TW) and epididymal sperm count (ESC) in normal diet group from 60 days (TW: 0.041 ± 0.0083 g; ESC: 1.575 ± 0.3079 × 105 cells) to 120 days (TW: 0.017 ± 0.0019 g; ESC: 0.22 ± 0.0189 × 105 cells); on the other hand, in goshajinkigan-including diet group, the dramatic recovery of these variables (TW: 0.10 ± 0.0062 g; ESC: 21.68 ± 1.705 × 105 cells) at 120 days, which is similarity to the normal spermatogenesis (TW: 0.098 ± 0.0024 g; ESC:19.8 ± 2.46 × 105 cells). These results suggest that busulfan-induced aspermatogenesis was irreversible unless receiving any medication. However, the supplementation of goshajinkigan can completely recover the regeneration of the injured semniferous epithelium, suggesting that goshajinkigan have a therapeutic effect on busulfan-induced aspermatogenesis. http://dx.doi.org/10.1016/j.jri.2014.09.027 Cadmium exposure increases the susceptibility to testicular autoimmunity in mice Y. Ogawa ∗ , M. Kuramasu, N. Qu, S. Hirai, S. Hayashi, N. Hatayama, M. Itoh Department of Anatomy, Tokyo Medical University, Japan Cadmium, one of various environmental toxicants, is known to suppress systemic immunity and to injure the testicular capillary endothelia with resultant
12
Abstracts for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20
necrosis of testicular tissues in mice and rats treated with high doses. Recently, it also became evident that cadmium can affect the integrity of the blood–testis barrier (BTB), the endocrine function of Leydig cells, apoptosis of germ cells and systemic immunity, even on treatment with a low dose that does not induce spermatogenic disturbance. Experimental autoimmune orchitis (EAO), i.e., an organ-specific autoimmunity of the testis, can be induced by repeated immunization with testicular antigens, and its pathology is characterized by lymphocytic inflammation and spermatogenic disturbance. In the present study, we investigated the morphological and functional changes of testes in mice treated with a low dose of cadmium chloride (CdCl2 ) and also examined its toxicity as to susceptibility to EAO. The results showed that exposure to 3 mg CdCl2/ kg body weight did not affect the spermatogenic state. However, the BTB at the tubuli recti and the rete testis, but not the seminiferous tubules, was slightly weakened, and intra-testicular mRNA expression of IL-6, TNF-␣ and IL-1 was significantly increased by the CdCl2 treatment. Furthermore, immunization with testicular antigens after the CdCl2 exposure significantly augmented the EAO severity. Therefore, exposure to a low dose of CdCl2 induces no significant disturbance of spermatogenesis, however, it does change the immunological microcircumstances in the testis, resulting in increased susceptibility to testicular autoimmunity. http://dx.doi.org/10.1016/j.jri.2014.09.028 Prevention of preterm labor by omega-3 polyunsaturated fatty acids and resolvin E3 E. Inoue ∗ , T. Nagamatsu, K. Kawana, T. Yamashita, Y. Osuga, T. Fujii Department of Obstetrics and Gynecology, Faculty of medicine, The University of Tokyo, Japan Introduction: Metabolites of omega-3 polyunsaturated fatty acids (PUFAs) have anti-inflammatory property, opposing inflammatory actions by omega-6-derived eicosanoids. Recently resolvin E3 is identified as a novel anti-inflammatory substance produced in the metabolic pathway of omega-3 PUFAs. Objective: This study aimed to investigate the possible role of omega 3 PUFAs in the regulation of intrauterine inflammation. We evaluated the relevance of omega-3 PUFAs and its metabolite, resolvin E3 to the pathology of preterm birth using a mouse model. Methods: Preterm birth was induced in mice by administering lipopolysaccharide (LPS) transcervically at day
15 pc. We examined the effect of resolvin E3 on the same model. Resolvin E3 was injected twice at the same time of LPS administration and at 6 h later to wild-type mice. Results: Preterm birth occurred in 87.5% in control mice and 56.3% in the mice after resolvin E3 injection (P = 0.03). The rate of fetal number left in the uterus was 34.9% in control mice and 65.3% in the mice with resolvin E3 injection (P < 0.01). Conclusions: We confirmed that omega-3 PUFAs suppress preterm birth induced by LPS. Our findings suggest that omega-3 PUFAs contribute to the prevention of inflammatory process caused by omega-6 PUFAs-derived eicosanoids induced by LPS injection. Anti-inflammatory metabolites of omega-3 PUFAs can be candidates for a new therapeutic medicine for preterm birth. http://dx.doi.org/10.1016/j.jri.2014.09.029 Interleukin-1 increases expression of tryptophan 2,3-dioxygenase and stimulates tryptophan metabolism in ectopic endomtetrial stromal cells Y. Urata ∗ , Y. Osuga, K. Koga, Y. Hirota, T. Hirata, M. Harada, M. Takamura, G. Izumi, T. Fujii The University of Tokyo, Department of Obstetrics and Gynecology, Japan Objective: Tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO) are enzymes that catalyze the first and rate-limiting step of tryptophan degradation (i.e. from tryptophan to kynurenine) and thereby regulate tryptophan levels. Decrease in tryptophan levels inhibits T cell function and contributes to immune tolerance. While IFN␥ increases IDO expression, a molecule that regulates TDO expression remains to be elucidated. We evaluated the effect of IL-1, a typical endometriosis-associated cytokine, on TDO expression in ectopic endometrial stromal cells (ESCs). Methods and results: ESCs were isolated from ovarian endometriomas and treated with IL-1. IL-1 (1 ng/ml) increased TDO mRNA expression to 6.3-fold of control but had no effect on IDO mRNA expression in ESCs. IL-1 increased TDO protein to 1.3-fold in ESCs and kynurenine production to 1.9-fold in the conditioned medium. TDO siRNA suppressed the increase in IL-1-induced kynurenine, IL-6 and IL-8 production. Conclusions: IL-1 increased TDO expression and stimulated tryptophan metabolism in ESCs. The IL-1-induced increase in TDO expression may enhance immune tolerance to promote endometriosis. http://dx.doi.org/10.1016/j.jri.2014.09.030