Dysfunction of neutrophil during parturition and proteome analysis of placenta and serum in cow

母牛分娩时中性粒细胞功能障碍及胎盘和血清蛋白质组分析

• 批准号: 15380199
• 负责人: INANAMI Osamu
• 金额: $ 9.09万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15380199/
• 关键词: bovine; neutrophil; signal transduction; two dimensional electrophoresis; bovine pregnancy related glycoprotein (bPAG); AKT; NADPH oxidase; keton body

The purpose of this project is to identify and characterize internal factors to reduce neutrophil functions after parturion in cows, since bovine mastitis sometimes occurs during parturition. In the experiments, the analysis of signal transduction pathways for activation of bovine neutrophils was performed. As important molecules for activation of NADPH oxidase in bovine neutrophils, cPKC, p38MAPK, ERK, PI3K and nPKC(pkcδ) were identified, but these factors did not changes during parturition in the cow. Furthermore, I tried to identify causative factors to reduce immune functions of cow after parturition by using two dimensional electrophoresis and other proteome technique. The proteome analysis showed that lactoferrin, transferrin, bovine pregnancy related glycoprotein (bPAG) and keton body in serum and placenta playd as suppressive factors for NADPH oxidase activation of bovine neutrophils. Moreover, to clarify the mechanism of this suppressive effects on neutrophil function, the effects of lactoferrin on LPS-or conA-induced expression of mRNA of INF-gamma, IL-1beta, TNF-alpha, IL12p4O and iNOS in mouse macrophage and feline monocyte was examined. The results indicated that the treatment of lactoferrin decreased LPS-or conA-induced expression of mRNA of INF-gamma and IL-1beta but did not affect that of TNF-alpha, IL12p4O and iNOS. The addition of lactoferrin to culture medium was showed to reduce differentiation form myeloblast to neutrophil in HL60 cells. These results suggest that lactoferrin was associated with regulation of inflammation and immune system mammary gland.

本项目的目的是确定和表征奶牛分娩后中性粒细胞功能降低的内部因素，因为牛乳房炎有时会发生在分娩期间。在实验中，对牛中性粒细胞活化的信号转导途径进行了分析。cPKC、p38 MAPK、ERK、PI 3 K和nPKC（pkcδ）是激活牛中性粒细胞NADPH氧化酶的重要分子，但这些因子在奶牛分娩过程中不发生变化。此外，本研究还利用双向电泳等蛋白质组学技术，对奶牛产后免疫功能下降的原因进行了初步研究。蛋白质组分析表明，血清和胎盘中的乳铁蛋白、转铁蛋白、牛妊娠相关糖蛋白（bPAG）和酮体对牛中性粒细胞NADPH氧化酶的激活起抑制作用。此外，为了阐明这种对中性粒细胞功能的抑制作用的机制，检测了乳铁蛋白对LPS或conA诱导的小鼠巨噬细胞和猫单核细胞中INF-γ、IL-1 β、TNF-α、IL 12 p4 O和iNOS的mRNA表达的影响。结果表明，乳铁蛋白处理降低了LPS或conA诱导的INF-γ和IL-1 β的mRNA表达，但对TNF-α、IL 12 p4 O和iNOS的mRNA表达没有影响。向培养基中添加乳铁蛋白显示减少HL 60细胞中从成髓细胞向中性粒细胞的分化。这些结果表明，乳铁蛋白与乳腺炎症和免疫系统的调节有关。

项目成果

A novel anticancer ribonucleoside, 1-(3-C-ethynyl-beta-D-ribo-pento-furanosyl)cytosine, enhances radiation-induced cell death in tumor cells.

一种新型抗癌核糖核苷 1-(3-C-乙炔基-β-D-核糖-戊呋喃糖基)胞嘧啶可增强肿瘤细胞中辐射诱导的细胞死亡。

• 发表时间: 2004
• 期刊: Radiation Research 162
• 作者: Inanami O.;Iizuka D.;Iwahara A.;Yamamori T.;Kon Y.;Asanuma T.;Matsuda A.;Kashiwakura I.;Kitazato K.;Kuwabara M.
• 通讯作者: Kuwabara M.

Induction of apoptosis through activation of SAPK/JNK followed by the expression of death receptor Fas in X-irradiated cells

在 X 射线照射的细胞中，通过 SAPK/JNK 的激活以及死亡受体 Fas 的表达来诱导细胞凋亡

• 发表时间: 2003
• 期刊: Journal of Radiation Research 44
• 作者: Kuwabara M.;Takahashi K.;Inanami O.
• 通讯作者: Inanami O.

Activation of c-kit by stem cell factor induces radioresistance to apoptosis through ERK-dependent expression of survivin in HL60 cells

• DOI: 10.1269/jrr.45.557
• 发表时间: 2004-12-01
• 期刊: JOURNAL OF RADIATION RESEARCH
• 影响因子: 2
• 作者: Hosseinimehr, SJ;Inanami, O;Kuwabara, M
• 通讯作者: Kuwabara, M

血管内皮細胞のシグナル伝達と活性酸素

血管内皮细胞的信号转导和活性氧

• 发表时间: 2005
• 作者: 丹波 光一;稲波 修
• 通讯作者: 稲波 修

Kuwabara M, Takahashi K, Inanami O.: "Induction of apoptosis through the activation of SAPK/JNK followed by the expression of death receptor Fas in X-irradiated cells"Journal of Radiation Research(Tokyo). 44(3) :. 203-209 (2003)

Kuwabara M、Takahashi K、Inanami O.：“通过 SAPK/JNK 的激活以及 X 射线照射的细胞中死亡受体 Fas 的表达来诱导细胞凋亡”放射线研究杂志（东京）。