Mechanism through which Kupffer cell-derived active oxidants mediate hepatoma cell injury

枯否细胞源活性氧化剂介导肝癌细胞损伤的机制

• 批准号: 07670613
• 负责人: KUROSE Iwao
• 金额: $ 1.6万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670613/
• 关键词: Hepatoma cell; Kupffer cell; NO; TNF-alpha; NF-kappaB; Adhesion molecules; Apoptosis; Laser scanning confocal microscope; 活性酸素; Nitric Oxide (NO); ミトコンドリア; アポトーシス

In 1995, we established the system which can determine productions of nitric oxide (NO) and TNF-alpha and apoptotic hepatoma cells with fragmented DNA and nuclear alterations. In 1996, by using laser scanning confocal microscope, we succeeded to visualize and analyze expression of mRNAs of inducible NO synthase (iNOS) and TNF-alpha. Investigations using these techniques revealed that coculture of Kupffer cells and hepatoma cells stimulate productions of NO and TNF-alpha, and these mediators released from Kupffer cells synergistically induce apoptosis of hepatoma cells. Inhibitor studies using antisense and sense oligodeoxynucleotides against mRNAs of iNOS and TNF-alpha, antibodies against adhesion molecules suggested that interactions via CD18 and ICAM-1 leads to activation and mediator production of Kupffer cells cocultured with hepatoma cells. We further established the method by which activated NF-kappaB can be visualized (Fluorescence in situ DNA-protein binding assay). By this assay, binding between CD18 and ICAM-1 have been demonstrated to lead to activation of NF-kappaB.

在1995年，我们建立了一个系统，可以确定一氧化氮（NO）和TNF-α的生产和凋亡肝癌细胞的DNA片段和核改变。1996年，我们利用激光扫描共聚焦显微镜，成功地观察和分析了诱导型一氧化氮合酶（iNOS）和肿瘤坏死因子-α（TNF-α）mRNA的表达。使用这些技术的研究表明，库普弗细胞和肝癌细胞的共培养刺激NO和TNF-α的产生，这些介质从库普弗细胞释放协同诱导肝癌细胞的凋亡。使用反义和正义寡脱氧核苷酸对iNOS和TNF-α mRNA的抑制剂研究，对粘附分子的抗体表明，通过CD 18和ICAM-1的相互作用导致与肝癌细胞共培养的库普弗细胞的激活和介体产生。我们进一步建立了激活的NF-κ B可以可视化的方法（荧光原位DNA-蛋白结合测定）。通过该测定，已经证明CD 18和ICAM-1之间的结合导致NF-κ B的活化。

项目成果

Kurose I, et al: "Rat Kupffer cell-derived nitric oxide woolulates" Gastroenterology. 109. 1958-1968 (1995)

Kurose I 等人：“大鼠 Kupffer 细胞衍生的一氧化氮羊毛”胃肠病学。

Kurose I, et al.: "Rat Kupffer cell-derived nitric oxide suppresses proliferation and induces apoptosis of syngeneic hepatoma cells." Gastroenterology. 111. 1058-1070 (1996)

Kurose I 等人：“大鼠 Kupffer 细胞来源的一氧化氮抑制同基因肝癌细胞的增殖并诱导细胞凋亡。”

Kurose I,et al.: "Rat Kupffer cell-derived nitric oxide suppresses proliferation and induces apoptosis of syngeneic hepatoma cells" Gastroenterology. 111. 1058-1070 (1996)

Kurose I 等人：“大鼠 Kupffer 细胞来源的一氧化氮抑制同基因肝癌细胞的增殖并诱导细胞凋亡”胃肠病学。

Kurose I,et al.: "Ethanol-induced oxidative stress in the liver." Alcholism : Clin Exp Res. 20. 77A-85A (1996)

Kurose I 等人：“乙醇诱导的肝脏氧化应激。”

Saito H, et al.: "Kupffer cell-mediated mitochondrial dysfunction in hepatoma cells by production of nitric oxide and the involvement of ICAM-1/LFA-1 molecules in their contact :" Int Immunol. 8. 1165-1172 (1996)

Saito H 等人：“库普弗细胞通过一氧化氮的产生以及 ICAM-1/LFA-1 分子参与其接触而介导肝癌细胞中的线粒体功能障碍：”IntImmunol。