Pathophysiology and Treatment of Endotoxin Shock

内毒素休克的病理生理学和治疗

• 批准号: 07557016
• 负责人: SHIMIZU Takao
• 金额: $ 8.45万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-07557016/
• 关键词: endotoxin; platelet activating factor; PAF; tyrosine phosphorylation; MAP kinase

Endotoxin causes systemic collapse, disseminated intravascular coagulation and lethality, by infection of gram-negative bacteria. Although many reports the production of cytokines, the precise mechanism of pathophysiological events remained obscure. By cell biology and molecular biology techniques, we obtained following results.(1) Elucidation of relation between endotoxin and PAF receptorWe established transgenic mice overxpressing PAF receptor. These mice showed severe symptoms of endotoxin shock and death by intravenous administration of small doses of LPS.The lethality was efficiently blocked by the use of PAF receptor antagonists, suggesting that PAF-PAFR is involved in the pathogenesis of endotoxin-induced lethality.(2) Elucidation of LPS action on macrophagesBy incubating of macrophages with bacterial endotoxin, PAFR mRNA was increased several fold after 4h. MAP kinase was also activated, and the activation related to the tyrosine phosphorylation of Raf-1 kinase.

内毒素通过革兰氏阴性菌感染导致全身崩溃、弥散性血管内凝血和致死。尽管许多报道了细胞因子的产生，但病理生理事件的确切机制仍然不清楚。通过细胞生物学和分子生物学技术，我们得到了以下结果： (1)内毒素与PAF受体关系的阐明我们建立了过表达PAF受体的转基因小鼠。这些小鼠通过静脉注射小剂量LPS​​表现出严重的内毒素休克和死亡症状。使用PAF受体拮抗剂可有效阻断致死性，这表明PAF-PAFR参与了内毒素诱导致死性的发病机制。(2 ) 阐明 LPS 对巨噬细胞的作用通过将巨噬细胞与细菌内毒素一起孵育，4 小时后 PAFR mRNA 增加数倍。 MAP激酶也被激活，该激活与Raf-1激酶的酪氨酸磷酸化有关。

项目成果

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Kishimoto, S.et al.：“IL-5 对人嗜酸性粒细胞上功能性血小板激活因子受体表达的调节。”

M. Nakamura et al: "Identification of two isoforms of mouse neuropeptide Y-Y1 receptor generated by alternative splicing, Isolatiom, genomic structure, and functional expression of the receptors." J, Biol. Chem.270. 30102-30108 (1995)

M. Nakamura 等人：“通过选择性剪接、分离、基因组结构和受体功能表达产生的小鼠神经肽 Y-Y1 受体的两种亚型的鉴定。”

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Mutoh, H. 等人：“人血小板活化因子受体基因对视黄酸和甲状腺激素的组织特异性反应是由两个不同的启动子引导的。”

Waga,I.: "Microtrap phosphorylation assay of mitogen-activated protein (MAP) kinases,to detect their activation by lipopolysaccharide." J.Immunol.Method. 190. 71-77 (1996)

Waga,I.：“丝裂原激活蛋白 (MAP) 激酶的 Microtrap 磷酸化测定，以检测脂多糖对它们的激活。”

Aoki, Y.et al.: "A radioreceptor binding assay for platelet-activating factor (PAF) receptor using membranes from CHO cells expressing human PAF receptor." J.Jmmunol. Method. 186. 225-231 (1995)

Aoki, Y. 等人：“使用表达人 PAF 受体的 CHO 细胞膜对血小板激活因子 (PAF) 受体进行放射性受体结合测定。”