豚鼠气道中NK1和NK2受体的鉴定。

Identification of both NK1 and NK2 receptors in guinea-pig airways.

作者信息

McKee K T, Millar L, Rodger I W, Metters K M

机构信息

Department of Pharmacology, Merck Frosst Centre for Therapeutic Research, Pointe-Claire, Dorval, Québec, Canada.

出版信息

Br J Pharmacol. 1993 Oct;110(2):693-700. doi: 10.1111/j.1476-5381.1993.tb13867.x.

DOI:10.1111/j.1476-5381.1993.tb13867.x

PMID:7694756

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2175923/

Abstract

NK1 and NK2 receptors have been characterized in guinea-pig lung membrane preparations by use of [125I-Tyr8]-substance P and [125I]-neurokinin A binding assays in conjunction with tachykinin-receptor selective agonists ([Sar9Met(O2)11]substance P for NK1 and [beta Ala8]neurokinin A (4-10) for NK2) and antagonists (CP-99,994 for NK1 and SR48968 for NK2). 2. The presence of high affinity, G-protein-coupled NK1 receptors in guinea-pig lung parenchymal membranes has been confirmed. The rank order of affinity for competing tachykinins was as predicted for an NK1 receptor: substance P = [Sar9Met(O2)11]substance P > substance P-methyl ester = physalaemin > neurokinin A = neurokinin B >> [beta Ala8]neurokinin A (4-10). The novel NK1 antagonist CP-99,994 has a Ki of 0.4 nM at this NK1 site. 3. In order to characterize [125I]-neurokinin A binding to guinea-pig lung, the number of [125I]-neurokinin A specific binding sites was increased 3-4 fold by purification of the parenchymal membranes over discontinuous sucrose gradients. The rank order of affinity determined for NK1- and NK2-receptor agonists and antagonists in competition for these sites showed that the majority (80%) of [125I]-neurokinin A specific binding was also to the NK1 receptor. 4. Under conditions where the guinea-pig lung parenchymal NK1 receptor was fully occupied by a saturating concentration of either [Sar9Met(O2)11]substance P (1 microM) or CP-99,994 (2.7 microM), residual [125I]-neurokinin A specific binding was inhibited in a concentration-dependent manner by both [beta Ala8]neurokinin A and SR48968. This result shows that the NK2 receptor is also present in these preparations. 5. Similar studies using guinea-pig tracheal membranes demonstrated that [125I]-neurokinin A specific binding was composed of a NK1-receptor component (60%), inhibited by both [Sar9Met(02)11]substance P and CP-99,994, and a significant NK2-receptor component, inhibited by both [beta Ala 8]neurokinin A andSR48968.6. In summary, these data demonstrate that guinea-pig lung parenchyma and guinea-pig trachea express both NK1 and NK2 receptors.

摘要

通过使用[125I-酪氨酸8]-P物质和[125I]-神经激肽A结合试验，并结合速激肽受体选择性激动剂（用于NK1的[Sar9Met(O2)11]P物质和用于NK2的[β丙氨酸8]神经激肽A(4-10)）以及拮抗剂（用于NKl的CP-99,994和用于NK2的SR48968），在豚鼠肺膜制剂中对NK1和NK2受体进行了表征。2. 已证实豚鼠肺实质膜中存在高亲和力、G蛋白偶联的NK1受体。对竞争性速激肽亲和力的排序与NK1受体的预测一致：P物质=[Sar9Met(O2)11]P物质>P物质甲酯=雨蛙肽>神经激肽A=神经激肽B>>[β丙氨酸8]神经激肽A(4-10)。新型NK1拮抗剂CP-99,994在该NK1位点的Ki为0.4 nM。3.为了表征[125I]-神经激肽A与豚鼠肺的结合，通过在不连续蔗糖梯度上纯化实质膜，[125I]-神经激肽A特异性结合位点的数量增加了3-4倍。在竞争这些位点时，对NK1和NK2受体激动剂及拮抗剂的亲和力排序表明，[125I]-神经激肽A特异性结合的大部分（80%）也是与NK1受体结合。4. 在豚鼠肺实质NK1受体被饱和浓度的[Sar9Met(O2)11]P物质(1μM)或CP-99,994(2.7μM)完全占据时[125I]-神经激肽A的残余特异性结合受到[β丙氨酸8]神经激肽A和SR48968浓度依赖性的抑制。该结果表明这些制剂中也存在NK2受体。5. 使用豚鼠气管膜的类似研究表明，[125I]-神经激肽A特异性结合由一个NK1受体成分（60%）组成，受[Sar9Met(02)11]P物质和CP-99,994抑制，以及一个显著的NK2受体成分，受[β丙氨酸8]神经激肽A和SR48968抑制。6. 总之，这些数据表明豚鼠肺实质和豚鼠气管均表达NK1和NK2受体。