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人外周血单个核细胞表达孤啡肽/痛敏肽,但不表达μ、δ或κ阿片受体。

Human peripheral blood mononuclear cells express nociceptin/orphanin FQ, but not mu, delta, or kappa opioid receptors.

作者信息

Williams John P, Thompson Jonathan P, McDonald John, Barnes Timothy A, Cote Tom, Rowbotham David J, Lambert David G

机构信息

Department of Cardiovascular Sciences, Pharmacology and Therapeutics Group, Division of Anaesthesia, Critical Care and Pain Management, University of Leicester, Leicester Royal Infirmary, Leicester, United Kingdom.

出版信息

Anesth Analg. 2007 Oct;105(4):998-1005, table of contents. doi: 10.1213/01.ane.0000278865.11991.9d.

Abstract

BACKGROUND

Expression of opioid receptors on peripheral blood mononuclear cells (PBMC) is controversial. These receptors are currently classified as classical (MOP/mu/mu, DOP/delta/delta and KOP/kappa/kappa) and nonclassical NOP (nociceptin/orphanin FQ; N/OFQ).

METHODS

In this volunteer study we probed for the expression of both classical and nonclassical opioid receptors using 1) radioligand binding, 2) specific antibody binding, and 3) polymerase chain reaction-based experimental paradigms.

RESULTS

Membranes prepared from PBMC from healthy volunteers did not bind either [3H]diprenorphine (a nonselective radioligand for classical opioid receptors) or [3H]N/OFQ. There was significant concentration-dependent binding of each radioligand to control tissues expressing recombinant MOP and NOP. In addition, using fluorescence-activated cell sorting paradigms, there was no binding of fluorescent naloxone or either of two MOP antibodies to whole PBMC, though fluorescent naloxone did bind to recombinant MOP (as a positive control). Using primers specific for classical and nonclassical opioid receptors, and RNA extracted from the PBMC of 10 healthy volunteers, we were also unable to detect MOP, DOP, and KOP transcripts. In contrast, NOP was detected in all samples.

CONCLUSIONS

Despite using several complementary experimental strategies, we failed to demonstrate protein for classical or nonclassical opioid receptors on PBMC from healthy volunteers. We detected NOP mRNA, suggesting low-density NOP expression on these immunocytes. It is possible that N/OFQ, produced by the PBMC itself, may be involved in the control of immune function.

摘要

背景

外周血单核细胞(PBMC)上阿片受体的表达存在争议。这些受体目前分为经典型(MOP/μ/μ、DOP/δ/δ和KOP/κ/κ)和非经典型NOP(孤啡肽/孤啡肽FQ;N/OFQ)。

方法

在这项志愿者研究中,我们使用1)放射性配体结合、2)特异性抗体结合和3)基于聚合酶链反应的实验范式来探究经典和非经典阿片受体的表达。

结果

从健康志愿者的PBMC制备的膜不与[3H]二丙诺啡(一种用于经典阿片受体的非选择性放射性配体)或[3H]N/OFQ结合。每种放射性配体与表达重组MOP和NOP的对照组织有显著的浓度依赖性结合。此外,使用荧光激活细胞分选范式,荧光纳洛酮或两种MOP抗体中的任何一种都不与完整的PBMC结合,尽管荧光纳洛酮确实与重组MOP结合(作为阳性对照)。使用针对经典和非经典阿片受体的特异性引物以及从10名健康志愿者的PBMC中提取的RNA,我们也未能检测到MOP、DOP和KOP转录本。相比之下,在所有样本中都检测到了NOP。

结论

尽管使用了几种互补的实验策略,但我们未能在健康志愿者的PBMC上证明经典或非经典阿片受体的蛋白表达。我们检测到了NOP mRNA,表明这些免疫细胞上存在低密度的NOP表达。PBMC自身产生的N/OFQ可能参与免疫功能的调节。

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