論文 - 桑山 岳人
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Severe maternal stress alters placental function, resulting in adipose tissue and liver dysfunction in offspring of mice 査読あり
Kondo R, Ozawa R, Satomi T, Funabayashi K, Iwata H, Kuwayama T, Shirasuna K.
Mol Cell Endocrinol 560 111814 2023年
記述言語：英語 掲載種別：研究論文（学術雑誌）
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PCR 法によるホロホロチョウ（Numida meleagris）の性判別 査読あり
南村昌孝・石田和真・黒澤 亮・高橋幸水・白砂孔明・岩田尚孝・小川 博・桑山岳人
日本家禽学会誌 59 ( J1 ) J1 - J5 2022年04月
担当区分：責任著者 記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：日本家禽学会
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Acetoacetate is a trigger of NLRP3 inflammasome activation in bovine peripheral blood mononuclear cells 査読あり
Onizawa Y, Katoh T, Miura R, Konda K, Noguchi T, Iwata H, Kuwayama T, Hamano S, Shirasuna K.
Vet Immunol Immunopathol 244 110370 2022年
記述言語：英語
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NLRP3 inflammasome is involved in testicular inflammation induced by lipopolysaccharide in mice 査読あり 国際誌
Sano M, Komiyama H, Shinoda R, Ozawa R, Watanabe H, Karasawa T, Takahashi M, Torii Y, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 87 e13527 2022年
記述言語：英語 掲載種別：研究論文（学術雑誌）
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ミナミコアリクイ（<i>Tamandua tetradactyla</i>）の同一ペア繁殖における 複数回妊娠中の血漿中性ステロイドホルモンの動態
日方 希保, 諸橋 菜々穂, 樽 舞帆, 鈴木 雄祐, 中村 智昭, 竹田 正裕, 桑山 岳人, 白砂 孔明
日本野生動物医学会誌 26 ( 4 ) 127 - 134 2021年12月
記述言語：日本語 出版者・発行元：日本野生動物医学会
<p>ミナミコアリクイ（<i>Tamandua tetradactyla</i>）は異節上目有毛目アリクイ科コアリクイ属に分類される哺乳類の一種である。コアリクイの計画的繁殖には，基礎的な情報の蓄積による繁殖生理の解明が必要である。これまでミナミコアリクイの妊娠期間中の血中ホルモン変動に関しては，1個体で1回分の妊娠期間についての報告がされているが，同一個体で複数回の妊娠期間中のホルモン変動に関する報告は存在しない。本研究では，同一雌個体のミナミコアリクイに対して長期間における経時的な採血（約1回/週）を実施し，同一雌雄ペアで合計6回の妊娠期間における血漿中プロジェステロン（P4）またはエストラジオール－17β（E2）濃度の測定を実施した。全6回の妊娠期間中のP4濃度測定の結果から，妊娠期間は156.8±1.7日（152～164日）と推定された。各時期のP4濃度は，妊娠前では0.6±0.1 ng/ml，妊娠初期（妊娠開始～出産100日以上前）では13.2±1.8 ng/ml，妊娠中期（出産50～100日前）では28.1±4.3 ng/ml，妊娠後期（出産日～出産50日前）では48.2±11.8 ng/mlであった。出産後のP4濃度は0.4±0.1 ng/mlと出産前から急激に低下した。血漿中E2濃度は妊娠初期から出産日に向けて徐々に増加した。また，妊娠期間前後で6回の発情周期様の変動がみられ，P4濃度動態から発情周期は45.5±2.4日（37～52日）と推定された。以上から，ミナミコアリクイの同一ペアによる複数回の妊娠中における血漿中性ステロイドホルモン動態を明らかにした。また，妊娠初期でP4濃度上昇が継続的なE2濃度上昇よりも先行して観察されたことから，P4濃度の連続的な上昇を検出することによって早期の妊娠判定が可能であることが示唆された。</p>
DOI： 10.5686/jjzwm.26.127
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IL1B triggers inflammatory cytokine production in bovine oviduct epithelial cells and induces neutrophil accumulation via CCL2 査読あり
Nakamura Y, Aihara R, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 85 e13365 2021年
記述言語：英語
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Uncaria tomentosa extract (AC-11) improves pregnancy hypertension together with suppression of sFlt-1 and sEng 査読あり
26 127 - 132 2021年
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos
KAGEYAMA Mio, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 67 ( 2 ) 141 - 147 2021年
記述言語：英語 出版者・発行元：The Society for Reproduction and Development
<p> The number of mitochondria in blastocysts is a potential marker of embryo quality. However, the molecular mechanisms governing the mitochondrial number in embryos are unclear. This study was conducted to investigate the effect of reduced mitochondrial reactive oxygen species (ROS) levels on mitochondrial biogenesis in porcine embryos. Oocytes were collected from gilt ovaries and activated to generate over 4 cell-stage embryos at day 2 after activation. These embryos were cultured in media containing either 0.1 μM MitoTEMPOL (MitoT), 0.5 μM Mitoquinol (MitoQ), or vehicle (ethanol) for 5 days to determine the rate of development to the blastocyst stage. The mitochondrial number in blastocysts was evaluated by real-time polymerase chain reaction (PCR). Five days after activation, the embryos (early morula stage) were subjected to immunostaining to determine the expression levels of NRF2 in the nucleus. In addition, the expression levels of <i>PGC1α</i> and <i>TFAM</i> in the embryos were examined by reverse transcription PCR. One day of incubation with the antioxidants reduced the ROS content in the embryos but did not affect the rate of development to the blastocyst stage. Blastocysts developed in medium containing MitoT had lower mitochondrial DNA copy numbers and ATP content, whereas MitoQ showed similar but insignificantly trends. Treatment of embryos with either MitoT or MitoQ decreased the expression levels of NRF2 in the nucleus and levels of <i>PGC1α</i> and <i>TFAM</i>. These findings indicate that reductions in mitochondrial ROS levels are associated with low mitochondrial biogenesis in embryos.</p>
DOI： 10.1262/jrd.2020-111
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HIRATA Yoshiki, KATSUKURA Yusuke, HENMI Yuka, OZAWA Ren, SHIMAZAKI Sayaka, KUROSAWA Akira, TORII Yasushi, TAKAHASHI Hironori, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 67 ( 4 ) 257 - 264 2021年
記述言語：英語 出版者・発行元：The Society for Reproduction and Development
<p> Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.</p>
DOI： 10.1262/jrd.2021-034
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos 査読あり
Kageyama M, Ito J, Shirasuna K, Kuwayama T, Iwata H.
J Reprod Dev 67 141 - 147 2021年
記述言語：英語 掲載種別：研究論文（学術雑誌）
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β-hydroxybutyrate suppresses NLRP3 inflammasome-mediated placental inflammation and lipopolysaccharide-induced fetal absorption 査読あり 国際誌
Hirata Y, Shimazaki S, Suzuki S, Henmi Y, Komiyama H, Kuwayama T, Iwata H, Karasawa T, Takahashi M, Takahashi H, Shirasuna K.
J Reprod Immunol 148 103433 2021年
記述言語：英語 掲載種別：研究論文（学術雑誌）
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カピバラ（<i>Hydrochoerus hydrochaeris</i>）の血液および糞を用いた発情周期の推定と妊娠判定方法の検討
諸橋 菜々穂, 佐野 宙矢, 加藤 孝典, 豊田 英人, 中村 智昭, 竹田 正裕, 桑山 岳人, 白砂 孔明
日本野生動物医学会誌 25 ( 4 ) 119 - 127 2020年12月
記述言語：日本語 出版者・発行元：日本野生動物医学会
<p> カピバラ（<i>Hydrochoerus hydrochaeris</i>）は南米に生息する最大の齧歯類である。カピバラの胎盤構造はヒトと類似しているという報告があり，実験動物への応用も期待できる。カピバラの計画的繁殖には繁殖生理の理解が重要だが，その情報は限られる。近年飼育現場等では，ハズバンダリートレーニング（HT）による低ストレスの採血や非侵襲的に採取可能な糞を用いた内分泌モニタリング方法の開発が進められている。本研究では，カピバラの血液および糞を用いた発情周期の特定や妊娠判定法について検討した。非妊娠個体でのHTによる採血とスメア採取を確立し，糞および血中性ステロイドホルモン濃度測定とスメア検査を行った。また，妊娠個体における糞中性ステロイドホルモン濃度測定を行った。非妊娠個体において血中プロジェステロン（P<sub>4</sub>）とエストロン3硫酸（E<sub>1</sub>S）濃度変動から明確な発情周期様の変動が3周期（平均11.7±0.9日）確認でき，血中E<sub>1</sub>S濃度変動は糞中エストラジオール（E<sub>2</sub>）濃度変動やスメアでの無核角化上皮細胞の出現と一致した。糞中E<sub>2</sub>濃度変動からも明確な周期的変動が15周期（平均8.5±0.3日）確認されたが，糞中P<sub>4</sub>濃度変動からは明確な周期性は確認できなかった。妊娠個体では，妊娠中期から出産直前における糞中P<sub>4</sub>およびE<sub>2</sub>濃度上昇が確認された。 以上から，血中P<sub>4</sub>やE<sub>1</sub>Sおよび糞中E<sub>2</sub>濃度測定とスメア検査からカピバラの発情周期が特定でき，糞中P<sub>4</sub>とE<sub>2</sub>濃度測定から妊娠判定実施の可能性が示唆された。</p>
DOI： 10.5686/jjzwm.25.119
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SANO Michiya, SHIMAZAKI Sayaka, KANEKO Yasuaki, KARASAWA Tadayoshi, TAKAHASHI Masafumi, OHKUCHI Akihide, TAKAHASHI Hironori, KUROSAWA Akira, TORII Yasushi, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 66 ( 3 ) 241 - 248 2020年
記述言語：英語 出版者・発行元：The Society for Reproduction and Development
<p> Maternal obesity is one of the major risk factors for pregnancy complications and is associated with low-grade chronic systemic inflammation due to higher levels of pro-inflammatory cytokines such as interleukin (IL)-1β. Pregnant women with obesity have abnormal lipid profiles, characterized by higher levels of free fatty acids, especially palmitic acid (PA). Previously, we reported that PA stimulated IL-1β secretion via activation of NLRP3 inflammasome in human placental cells. These observations led us to hypothesize that higher levels of PA induce NLRP3 inflammasome activation and placental inflammation, resulting in pregnancy complications. However, the effects of PA on NLRP3 inflammasome during pregnancy <i>in vivo</i> remain unclear. Therefore, PA solutions were administered intravenously into pregnant mice on day 12 of gestation. Maternal body weight was significantly decreased and absorption rates were significantly higher in PA-injected mice. The administration of PA significantly increased IL-1β protein and the mRNA expression of NLRP3 inflammasome components (<i>NLRP3</i>, <i>ASC</i>, and caspase-1) within the placenta. In murine placental cell culture, PA significantly stimulated IL-1β secretion, and this secretion was suppressed by a specific NLRP3 inhibitor (MCC950). Simultaneously, the number of macrophages/monocytes and neutrophils, together with the mRNA expression of these chemokines increased significantly in the placentas of PA-treated mice. Treatment with PA induced ASC assembling and IL-1β secretion in macrophages, and this PA-induced IL-1β secretion was significantly suppressed in NLRP3-knockdown macrophages. These results indicate that transient higher levels of PA exposure in pregnant mice activates NLRP3 inflammasome and induces placental inflammation, resulting in the incidence of absorption.</p>
DOI： 10.1262/jrd.2020-007
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KOBAYASHI Mitsuru, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 66 ( 6 ) 539 - 546 2020年
記述言語：英語 出版者・発行元：The Society for Reproduction and Development
<p> We examined the effect of ploidy on mitochondrial DNA (mtDNA) copy number in embryos and the amount of cell-free mitochondrial and nucleic DNA content (cf-mtDNA and cf-nDNA) in spent culture medium (SCM). Oocytes collected from the ovaries were matured, activated, incubated in medium containing cycloheximide (CHX) or CHX and cytochalasin B (CB) for 4.5 h to produce haploid or diploid embryos (H-group and D-group embryos). These embryos were cultured for 7 days, and the blastocysts and SCM were examined. The amount of mtDNA and nDNA was determined by real-time PCR. The rate of development to the blastocyst stage was higher for the D-group than for the H-group. Moreover, D-group blastocysts had less mtDNA compared to the H-group blastocysts. After activation, the mitochondrial content was constant before the blastocyst stage in D-group embryos, but increased earlier in H-group embryos. The amount of cf-mtDNA in the SCM of D-group blastocysts was greater than that of H-group blastocysts. However, when the cf-mtDNA in the SCM of 2 cell-stage embryos (day 2 post-activation) was examined, the amount of cf-mtDNA was greater in the H-group than in the D-group embryos. When D-group embryos were cultured for 7 days, a significant correlation was observed between the total cell number of blastocysts and cf-nDNA content in the SCM. Hence, although careful consideration is needed regarding the time point for evaluating mtDNA content in the embryos and SCM, this study demonstrates that mtDNA in the embryos and SCM was affected by the ploidy of the embryos.</p>
DOI： 10.1262/jrd.2020-097
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Interferon tau regulates cytokine production and cellular function in human trophoblast cell line. 査読あり
Tanikawa N, Seno K, Kawahara-Miki R, Kimura K, Matsuyama S, Iwata H, Kuwayama T, Shirasuna K
J Interferon & Cytokine Research 37 456 - 466 2017年10月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Advanced glycation end products and lipopolysaccharide stimulate interleukin-6 secretion via RAGE/TLR4-NF-kB-ROS pathways and resveratrol attenuates these inflammatory responses in mouse macrophages.
Ohtsu A, Shibutani Y, Seno K, Iwata H, Kuwayama T, Shirasuna K.
Experimental and Therapeutic Medicine 14 4364 - 4370 2017年08月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Age-associated mRNA expression changes in bovine endometrial cells in vitro. 査読あり
Tanikawa N, Ohtsu A, Kawahara-Miki R, Kimura K, Matsuyama S, Iwata H, Kuwayama T, Shirasuna K
Reprod Biol Endocrinol 63 2017年08月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Advanced glycation end products regulate interleukin-1β production in human placenta. 査読あり
Seno K, Sase S, Ozeki A, Takahashi H, Ohkuchi A, Suzuki H, Matsubara S, Iwata H, Kuwayama T, Shirasuna K
J Reprod Dev 63 401 - 408 2017年05月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Palmitic acid stimulates interleukin-8 via the TLR4/NF-kB/ROS pathway and induces mitochondrial dysfunction in bovine oviduct epithelial cells. 査読あり
Ohtsu A, Tanaka H, Seno K, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 77 2017年02月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Promotion of glucose utilization by insulin enhances granulosa cell proliferation and developmental competence of porcine oocyte growth in vitro. 査読あり
Itami N, Munakata Y, Shirasuna K, Kuwayama T, Iwata H
Zygote 25 65 - 74 2016年12月
記述言語：英語 掲載種別：研究論文（学術雑誌）