DEVELOPMENT OF NOVEL ASSAY SYSTEM FOR ANTI-ALLERGIC COMPOUNDS FROM NATURAL SOURCES AND APPLICATION

天然来源抗过敏化合物新型检测系统的开发及应用

基本信息

批准号：
17510185
负责人：
ISHIGURO Kyoko
金额：
$ 2.37万
依托单位：
Mukogawa Women's University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17510185/
关键词：
Blood flow monitoring Hen-egg white lysozyme iNOS KO mouse inducible NO synthase (iNOS)e NOS (6) COX-1 and 2 Allergy preventive medicines in vivo assay system in vivoアッセイ法 NO COX-1、2 PGI_2

项目摘要

Our in vivo assay system developed to search for allergy-preventive substances, assesses the blood flow decrease in tail vein microcirculation of mice subjected to sensitization with hen-egg white lysozyme (HEL). The blood flow decrease appears to be regulated by various factors such as nitric oxide (NO), thromboxane (TX) A2, prostacyclin (PGI2) and endothelin (ET)-1 together with cyclooxygenase (COX)-1, 2, and inducible nitric oxide synthase (iNOS).In this study, we examined in detail the roles of iNOS in this assay system using an iNOS knockout (KO) mouse. We found that the blood flow decrease in the HEL-sensitized iNOS KO mice was slightly weaker than that in their wild type (WT) mice. This blood flow decrease was not affected by a selective COX-1 inhibitor, a COX-2 inhibitor and a PGI2 agonist unlike the case of the WT mice. However, it was inhibited by a nonselective NOS inhibitor, a specific TXA2 synthase inhibitor and a specific ET-1 receptor blocker as in the case of the WT mice. The present results indicate that the blood flow decrease occurs via two pathways; one is an iNOS-independent response involving TXA2 and ET-1, and the other is an iNOS-dependent response involving COX-1, COX-2 and PGI2. Constitutive nitric oxide synthase (cNOS) appears to play some roles in the blood flow decrease and iNOS acts as an exacerbation factor. Our method using HEL-sensitized should be useful for searching for agents that can prevent allergy via new mechanisms.The other hand we also demonstrated allergy-preventive activity of extract of Populus sieboldii, Xanthorrhoea hastilis and Lonicera japonica using this in vivo assay system. By bioassay-directed fractionation of these plans, new allergy-preventive compounds were isolated.

我们开发的体内检测系统是为了寻找过敏预防物质，评估用鸡蛋清溶菌酶 (HEL) 致敏的小鼠尾静脉微循环血流量的减少情况。血流量减少似乎受到多种因素的调节，例如一氧化氮 (NO)、血栓素 (TX) A2、前列环素 (PGI2) 和内皮素 (ET)-1 以及环氧合酶 (COX)-1、2 和诱导型硝酸盐氧化物合酶 (iNOS)。在这项研究中，我们使用 iNOS 敲除 (KO) 小鼠详细检查了 iNOS 在该检测系统中的作用。我们发现 HEL 致敏的 iNOS KO 小鼠的血流量减少略弱于野生型 (WT) 小鼠。与 WT 小鼠的情况不同，这种血流量减少不受选择性 COX-1 抑制剂、COX-2 抑制剂和 PGI2 激动剂的影响。然而，与 WT 小鼠一样，它受到非选择性 NOS 抑制剂、特定 TXA2 合酶抑制剂和特定 ET-1 受体阻滞剂的抑制。目前的结果表明血流量减少通过两个途径发生：一种是涉及 TXA2 和 ET-1 的 iNOS 独立反应，另一种是涉及 COX-1、COX-2 和 PGI2 的 iNOS 依赖性反应。组成型一氧化氮合酶 (cNOS) 似乎在血流量减少中发挥一定作用，而 iNOS 则充当恶化因素。我们使用 HEL 致敏的方法应该有助于寻找可以通过新机制预防过敏的药物。另一方面，我们还使用这种体内测定系统证明了西氏杨、Xanthorrhoea hastilis 和忍冬提取物的过敏预防活性。通过对这些计划进行生物测定指导的分离，分离出新的过敏预防化合物。