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Study on the function of endogenous opioid and substance P at inflammation and pain

内源性阿片类药物和P物质对炎症和疼痛的作用研究

基本信息

批准号：
10671443
负责人：
NISHIMURA Kinya
金额：
$ 1.47万
依托单位：
Juntendo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671443/
关键词：
spinorphin enkephalin aminopeptidase bradykinin Ni-NTA dipeptydylaminopeptidase substance P substance P receptor Spinorphin Leuhistin PMN

项目摘要

We synthesized spinorphin analogous and assayed their inhibitoriy activity toward dipeptidyl peptidase III (DPPIII) among enkephalin degrading enzymes. VVYPW, an N-terminal and C-terminal truncated form of spinorphin, exhibited more potent inhibitory activity.This results indicated that VVYPW had amore effective structure of expression of inhibitory activity toward DPPIII.We attempted to characterize nociceptive sensory fibers into three types, and examined the mode of action of spinorphin-induced analgesia, in comparison to morphine-induced one.So, spinorphin completely blocked 2-metyl-thioadenosine induced responsed, but morphine did not. On the other hand, morphine-induced blockade of bradykinin responses was attenuated by pertussis toxin treatment, while the spinorphin's ones was not. Thus it is suggested that spinorphin has wide spectrum of analgesia which covers the blockade of nociception insensitive to morphine.Regarding to purification of human substance P receptor, we have developed a recombinant Substance P receptor with 6xHis epitope at its N-terminus. The 6xHis tag does not interfere with receptor's ability to bind ligands the 6XHis-SPR can be purified over the Ni-NTA column. This purification procedure constitutes an important step toward obtaining large quantities of homogeneous SPR for structural studies.

我们合成了类似于spinorphin的物质，并测定了它们对脑啡肽降解酶中二肽基肽酶III （DPPIII）的抑制活性。VVYPW是spinorphin的n端和c端截断形式，表现出更强的抑制活性。结果表明，VVYPW具有有效的抑制DPPIII活性的表达结构。我们试图将伤害性感觉纤维分为三种类型，并检查了脊髓啡诱导的镇痛作用模式，与吗啡诱导的镇痛作用模式进行了比较。因此，脊髓啡完全阻断了2-甲基硫腺苷诱导的反应，而吗啡则没有。另一方面，吗啡诱导的缓激肽阻滞反应被百日咳毒素处理减弱，而脊髓啡的阻滞反应则没有。提示脊髓啡具有广谱的镇痛作用，可阻断对吗啡不敏感的痛觉。在纯化人P物质受体方面，我们开发了一种重组P物质受体，其n端具有6xHis表位。6xHis标签不干扰受体结合配体的能力，6xHis - spr可以通过Ni-NTA柱纯化。该纯化过程是获得大量均匀SPR用于结构研究的重要一步。