Basic research about pathophysiology of encephalopathy with hemolytic uremic syndrome

本研究采用人脑内皮细胞血脑屏障模型，对产志贺毒素肠出血性大肠杆菌感染急性脑病（HUS脑病）合并溶血性尿毒症综合征（HUS）的病理生理进行基础研究。(1)考虑Stx刺激人脑内皮细胞皮肤细胞中IL-8、MCP-1等趋化因子的表达，以及趋化因子对白细胞粘附/移动能力或Gb3表达能力的影响。(2)该细胞培养体系采用抗il -8抗体和抗mcp -1抗体的抗趋化因子抗体，考虑了白细胞粘附/移位能力、Gb3表达能力和Stx的参与。(3)Stx考虑了其对参与趋化因子表达的传递因子NF-kappa B的影响。作为制备阶段的血脑屏障模型，建立人脑内皮细胞培养体系，通过测定培养液检测趋化因子的升高，以Stx刺激下IL-8、MCP-1等趋化因子为导向。趋化因子目前也在本会计年度考虑其对白细胞粘附/移动能力或Gb3表达能力的影响。提取Stx刺激培养的细胞总RNA， northern blot法检测IL-8和MCP-1的表达是否显著。目前使用抗il -8抗体和抗mcp -1抗体的抗趋化因子抗体的实验仍在继续。Stx刺激对传递因子nf - κ B的激活也进行了研究。此外，在本财政年度，作为原始模型的人脑内皮细胞的收购也很困难，正在与相关组织和公司进行谈判。对某一研究课题有意义的实验数据在三年内尚未得出。

In this research, fundamental research on the pathophysiology of the acute encephalopathy (HUS encephalopathy) accompanying the hemolytic uremic syndrome (HUS) by Shiga toxin-producing enterohemorrhagic Escherichia coli infection is done using the blood-brain barrier model by the human cerebral endothelial cell.(1)Chemokine expression of IL-8,MCP-1, etc. in the skin cell in the human cerebral endothelial cell by Stx stimulus, and the influence which chemokine has on leukocyte adhesion/shift ability or Gb3 expression ability are considered.(2)This cell culture system considers participation of leukocyte adhesion/shift ability, Gb3 expression ability, and Stx using the anti-chemokine antibody of anti-IL-8 antibody and anti-MCP-1 antibody.(3)Stx considers the influence, which it has on transfer factor NF-kappa B that participates in chemokine expression. As the blood-brain barrier model in a preparation stage, the cultivation system of the human cerebral endothelial cell were established, and the rise of chemokine was checked for guiding chemokine, such as IL-8 in a Stx stimulus, and MCP-1, by measurement of culture solution.Chemokine is considering the influence which it has on leukocyte adhesion/shift ability or Gb3 expression ability also in the current fiscal year now.Moreover, total RNA was extracted from the cell cultivated by Stx stimulus, the northern blot method of IL-8 and MCP-1 was performed, and significant expression was checked.The experiment using the anti-chemokine antibody of anti-IL-8 antibody and anti-MCP-1 antibody is under continuation now.It is under examination also about the transfer factor NF-kappa B activation by Stx stimulus. Also in the current fiscal year, acquisition of the human cerebral endothelial cell, which is an original model, is difficult, and is negotiating with the related organization and the company.The significant experiment data to a research subject are not drawn yet in three years.