The significance of Osteopontin in development of necrosis, regeneration, fibrosis and carcinogenesis in the liver : Evaluation using osteopontin transgenic mice.

骨桥蛋白在肝脏坏死、再生、纤维化和癌变发展中的重要性：使用骨桥蛋白转基因小鼠进行评估。

• 批准号: 14570505
• 负责人: MOCHIDA Satoshi
• 金额: $ 2.5万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570505/
• 关键词: Osteopontin; Transgenic Mouse; Cytokine; Liver Necrosis; Autoimmune Hepatitis; Lymphcytes; Fulminant Hepatitis; Extrahepatic Lesions

Osteopontin, an extracellular matrix with RGD sequence, is shown to act as a cytokine essential for initiation of Th1 immune response. Although osteopontin expression is minimal in normal liver, up-regulation of the expression develops in Kupffer cells, macrophages, stellate cells and hepatocytes both in mice and human with liver injury. To clarify the role of osteopontin in the development of liver injury, we made transgenic mice expressing osteopontin exclusively in hepatocytes using the vector containing human serum amyloid P component promoter. In the present study, we evaluated immunological and pathological features of these mice in relation to autoimmune hepatitis.Marked lymphocyte infiltration developed in the periportal areas of the liver later than 12 weeks of age both in male and female mice. These cells were positive for both CD8 and HLA-DR, suggesting that cytotoxic T cells infiltrated in the liver. In these mice, focal necrosis was found in the hepatic lobules. Also, lymp … More hocyte infiltration was observed in the spleen, lung, pancreas and salivary gland, and marked splenomegaly was found later than 24 weeks. Anti-nuclear antibodies in sera assessed by fluorescence immunostaining were positive in 50% of rats between 8 and 48 weeks. Gene expression profile was evaluated in the liver using DNA microarray in mice at 12 weeks. Caplain mRNA expressions were decreased in the liver of transgenic mice compared to non-transgenic mice, while mRNA expressions of metallothionein and glutathione-s-transferase were up-regulated. Serum levels of IFN-γ and IL-10 did not differ between transgenic and non-transgenic mice. The levels increased following intravenous injection of concanavalin-A in both groups. However, IFN-γ level was much more increased in transgenic mice compared to non-transgenic mice, while increase in IL-10 level was less in transgenic mice than in non-transgenic mice. The extent of liver necrosis following concanavalin-A administration was greater in transgenic mice than in non-transgenic mice.Transgenic mice expressing osteopontin in hepatocytes may be a useful model of autoimmune hepatitis, since anti-nuclear antibody was positive in the serum and focal necrosis accompanied by cytotoxic T lymphocyte infiltration developed in the liver. In these mice, mRNA expressions of radical scavenger and detoxifing enzyme were up-regulated prominently in the liver. Also, the mice showed deranged balance in Th1/Th2 cytokine profiles following concanavalin-A administration. Thus, the transgenic mice would be useful tin the study of osteopontin in the development of liver injury in vivo. Less

骨桥蛋白是一种含有RGD序列的细胞外基质，是Th 1免疫应答启动所必需的细胞因子。尽管骨桥蛋白在正常肝脏中的表达是最小的，但在肝损伤的小鼠和人的枯否细胞、巨噬细胞、星状细胞和肝细胞中表达上调。为了阐明骨桥蛋白在肝损伤发展中的作用，我们使用含有人血清淀粉样蛋白P组分启动子的载体制作了仅在肝细胞中表达骨桥蛋白的转基因小鼠。在本研究中，我们评估了这些小鼠与自身免疫性肝炎相关的免疫学和病理学特征。雄性和雌性小鼠在12周龄后肝脏门静脉周围区域均出现明显的淋巴细胞浸润。这些细胞对CD 8和HLA-DR均呈阳性，表明细胞毒性T细胞在肝脏中浸润。在这些小鼠中，在肝小叶中发现局灶性坏死。另外，Lymp ...更多信息 在脾、肺、胰腺和唾液腺中观察到红细胞浸润，并且在24周后发现显著的脾肿大。在8和48周之间，50%的大鼠通过荧光免疫染色评估血清中的抗核抗体呈阳性。在12周时使用DNA微阵列评估小鼠肝脏中的基因表达谱。与非转基因小鼠相比，转基因小鼠肝脏中的Caplain mRNA表达降低，而金属硫蛋白和谷胱甘肽-S-转移酶的mRNA表达上调。转基因小鼠和非转基因小鼠的血清IFN-γ和IL-10水平没有差异。在两组中，静脉注射刀豆球蛋白-A后水平均升高。然而，与非转基因小鼠相比，转基因小鼠中IFN-γ水平显著升高，而转基因小鼠中IL-10水平的升高低于非转基因小鼠。转基因小鼠肝细胞内表达骨桥蛋白，血清抗核抗体阳性，肝细胞出现局灶性坏死，并伴有细胞毒性T淋巴细胞浸润，可作为自身免疫性肝炎的模型。在这些小鼠中，自由基清除剂和解毒酶的mRNA表达在肝脏中显著上调。此外，伴刀豆球蛋白A给药后，小鼠的Th 1/Th 2细胞因子谱出现紊乱。因此，转基因小鼠将有助于研究骨桥蛋白在肝损伤中的作用。少

项目成果

Fujimori K, Mochida S, Matsui A, Ohno A, Fujiwara K.: "Possible mechanisms of evaluation of serum transaminase levels during interferon-beta therapy in chronic hepatitis C patients"Journal of Gastroenterology. 37. 40-46 (2002)

Fujimori K、Mochida S、Matsui A、Ohno A、Fujiwara K.：“慢性丙型肝炎患者干扰素β治疗期间血清转氨酶水平评估的可能机制”胃肠病学杂志。

Matsui A, Ohno A, Nagoshi S, Fujiwara K.: "Plasma Osteopontin Levels in Patients with Fulminant Hepatitis"Hepatology Research. (印刷中). (2004)

Matsui A、Ohno A、Nagoshi S、Fujiwara K.：“暴发性肝炎患者的血浆骨桥蛋白水平”肝病学研究（2004 年）。

Takahashi M, Matsui A, Inao M, Mochida S, Fujiwara K.: "ERK/MAPK-dependent PI3K/Akt phosphorylation through VEGFR-1 after VEGF stimulation in activated Hepatic stellate cells"Hepatology Research. 26. 233-236 (2003)

Takahashi M、Matsui A、Inao M、Mochida S、Fujiwara K.：“激活的肝星状细胞中 VEGF 刺激后，ERK/MAPK 依赖性 PI3K/Akt 通过 VEGFR-1 磷酸化”肝病学研究。

Takahashi M, Matsui A, Inao M, Mochida S, Fujiwara K.: "ERK/MAPK-Dependent P13K/Akt Phosphorylation through VEGFR-1 after VEGF Stimulation in Activated Hepatic Stellate Cells."Hepatologoy Res. 26. 233-236 (2003)

Takahashi M、Matsui A、Inao M、Mochida S、Fujiwara K.：“激活的肝星状细胞中 VEGF 刺激后通过 VEGFR-1 进行 ERK/MAPK 依赖性 P13K/Akt 磷酸化。”肝病学研究。

Matsui A, Mochida S, Ohno A, Nagoshi S, Hirose T, Fujiwara K.: "Plasma Osteopontin Levels in Patients with Fulminant Hepatitis."Hepatology Res. (in press).

Matsui A、Mochida S、Ohno A、Nagoshi S、Hirose T、Fujiwara K.：“暴发性肝炎患者的血浆骨桥蛋白水平。”肝病学研究。