THE ROLE OF GLOMERULAR ENDTHELIAL CELLS IN THE PATHOGENESIS OF GLOMERULONEPHRITIS.

肾小球内皮细胞在肾小球肾炎发病机制中的作用。

• 批准号: 04670573
• 负责人: KAKIZAKI Yoshiki
• 金额: $ 1.34万
• 依托单位: HIROSAKI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670573/
• 关键词: Glomerulonephritis; Monocyte; Macrophage; Glomerular endothelial cell; Monocyte chemoattractant protein-1; Protein kinase; gene expression; MCP-1; 転写因子NFκB; 単球遊走因子; メサンギウム細胞; 細胞外基質成分; 接着分子; 単球

To explore the role of glomerular endothelial cells (GEN) in the pathogenesis of glomerulonephritis, the in vitro production of monocyte chemoattractant protein-1 (MCP-1) by bovine GEN was determined by chemotaxis assay, Northern blot analysis, and immunocytochemistry. Monocyte chemotactic activity of GEN-conditioned media was detectable by a chemotaxis assay using human peripheral blood monocytes. Exposure to human recombinant interleukin-1beta (IL-1beta) and phorbol myristate acetate (PMA) significantly increased the chemotactic activity of GEN-conditioned media. A checcurboard analysis showed that the response of monocytes to GEN-conditioned media was truly chemotactic. Immunoadsorption with a monoclonal antibody to human MCP-1 reduced the chemotactic activity of GEN-conditioned media by 85%. Northern blot analysis revealed that MCP-1 mRNA was constitutively expressed by GEN and that IL-1beta and tumor necrosis factor-alpha (TNF-alpha) increased MCP-1 mRNA levels in a dose- and time … More -dependent manner. PMA also induced an increase in MCP-1 mRNA levels that was suppressed by the protein kinase C inhibitors staurosporine and H-7, whereas dibutyryl cyclic AMP and forskolin had minimal effects, indicating involvement of protein kinase C.However, MCP-1 mRNA expression induced by IL-1beta and TNF-alpha was suppressed by the tyrosine kinase inhibitor genistein, not by staurosporine, H-7, or the protein kinase A inhibitor H-89, suggesting importance of the tyrosine kinase activation on the cytokine-induced MCP-1 gene expression. Dexamethasone had a small inhibitory effect on constitutive and PMA-induced MCP-1 mRNA expression, but no effect on the induction by TNF-alpha.By immunoperoxidase staining with an anti-MCP-l monoclonal antibody, MCP-1 protein was detected in untreated GEN and increased by exposure to PMA,correspondeng to mRNA expression. These results demonstrate the production of MCP-1 by GEN at gene and protein levels as well as bioactivity, and suggest that GEN may participate in the development of glomerulonephritis through the production of MCP-1. Less

为探讨肾小球内皮细胞（glomerular endothelial cells， GEN）在肾小球肾炎发病中的作用，采用趋化性实验、Northern blot分析和免疫细胞化学方法检测牛内皮细胞体外产生单核细胞趋化蛋白-1 （MCP-1）的情况。利用人外周血单核细胞进行趋化试验，检测gen条件培养基的单核细胞趋化活性。暴露于人重组白细胞介素-1 β (il -1 β)和肉豆蔻酸酯phorbol acetate （PMA）显著增加gen条件培养基的趋化活性。棋盘分析显示单核细胞对gen条件培养基的反应是真正的趋化。人MCP-1单克隆抗体免疫吸附使gen条件培养基的趋化活性降低85%。Northern blot分析显示，GEN可组成性表达MCP-1 mRNA， il -1 β和肿瘤坏死因子α (tnf - α)以剂量和时间依赖性方式增加MCP-1 mRNA水平。PMA还诱导MCP-1 mRNA水平的增加，而MCP-1 mRNA水平被蛋白激酶C抑制剂staurosporine和H-7抑制，而二丁基环AMP和forskolin的影响很小，表明与蛋白激酶C有关。然而，il -1 β和tnf - α诱导的MCP-1 mRNA表达被酪氨酸激酶抑制剂genistein抑制，而不是被staurosporine， H-7或蛋白激酶A抑制剂H-89抑制。提示酪氨酸激酶激活对细胞因子诱导的MCP-1基因表达的重要性。地塞米松对组成型和pma诱导的MCP-1 mRNA表达有微弱抑制作用，但对tnf - α诱导的MCP-1 mRNA表达无影响。通过抗MCP-1单克隆抗体免疫过氧化物酶染色，在未处理的GEN中检测到MCP-1蛋白，暴露于PMA后MCP-1蛋白表达增加，与mRNA表达相对应。这些结果证实了GEN在基因和蛋白水平以及生物活性水平上产生MCP-1，提示GEN可能通过产生MCP-1参与肾小球肾炎的发生。少

项目成果

柿崎良樹: "培養糸球体内皮細胞による単求遊走因子産生の検討" 日本小児腎臓病学会雑誌. 6. 332- (1993)

Yoshiki Kakizaki：“培养肾小球内皮细胞产生单一趋化因子的研究”日本小儿肾病学会杂志 6. 332- (1993)。

YOSHIKI KAKIZAKI: "PRODUCTION OF MONOCYTE CHEMOTACTIC FACTOR BY GLOMERULAR ENDOTHELIAL CELLS IN CULTURE." JAPANESE JOURNAL OF PEDIATRIC NEPHROLOGY. VOL.6. 332 (1993)

YOSHIKI KAKIZAKI：“培养中肾小球内皮细胞产生单核细胞趋化因子。”