Zhang Lan, Stamer Ulrike M, Moolan-Vadackumchery Robin, Stüber Frank
Department of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, Switzerland.
Department for BioMedical Research, University of Bern, 3008 Bern, Switzerland.
Cells. 2024 Dec 20;13(24):2111. doi: 10.3390/cells13242111.
The nociceptin receptor (NOP) and nociceptin are involved in the pathways of pain and inflammation. The potent role of nuclear factor-κB (NFκB) in the modulation of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β on the nociceptin system in human THP-1 cells under inflammatory conditions were investigated. Cells were stimulated without/with phorbol-myristate-acetate (PMA), TNF-α, IL-1β, or PMA combined with individual cytokines. To examine NFκB's contribution to the regulation of the nociceptin system, PMA-stimulated cells were treated with NFκB inhibitor BAY 11-7082, JSH-23, or anacardic acid before culturing with TNF-α or IL-1β. and prepronociceptin () mRNA were quantified by RT-qPCR; cell membrane NOP and intracellular nociceptin protein levels were measured by flow cytometry. Phosphorylation and localization of NFκB/p65 were determined using ImageStream. PMA + TNF-α decreased mRNA compared to stimulation with PMA alone, while PMA + IL-1β did not. BAY 11-7082 and JSH-23 reversed the repression of by PMA + TNF-α. TNF-α and IL-1β attenuated PMA's upregulating effects on . None of the inhibitors preserved the upregulation of in PMA + TNF-α and PMA + IL-1β cultures. TNF-α strongly mediated the nuclear translocation of NFκB/p65 in PMA-treated cells, while IL-1β did not. Proinflammatory cytokines suppressed and mRNA in PMA-induced human THP-1 cells. NFκB signaling seems to be an important regulator controlling the transcription of NOP. These findings suggest that the nociceptin system may play an anti-inflammatory role during immune responses.
孤啡肽受体（NOP）和孤啡肽参与疼痛和炎症通路。研究了核因子-κB（NFκB）在炎症条件下对人THP-1细胞中肿瘤坏死因子-α（TNF-α）和白细胞介素（IL）-1β调节孤啡肽系统的重要作用。用/不用佛波酯-肉豆蔻酸酯-乙酸酯（PMA）、TNF-α、IL-1β或PMA与单个细胞因子联合刺激细胞。为了研究NFκB对孤啡肽系统调节的作用，在用TNF-α或IL-1β培养之前，用NFκB抑制剂BAY 11-7082、JSH-23或没食子酸处理PMA刺激的细胞。通过逆转录定量聚合酶链反应（RT-qPCR）对前体孤啡肽原（）mRNA进行定量；通过流式细胞术测量细胞膜NOP和细胞内孤啡肽蛋白水平。使用图像流式细胞仪确定NFκB/p65的磷酸化和定位。与单独用PMA刺激相比，PMA + TNF-α降低了mRNA水平，而PMA + IL-1β则没有。BAY 11-7082和JSH-23逆转了PMA + TNF-α对的抑制作用。TNF-α和IL-1β减弱了PMA对的上调作用。在PMA + TNF-α和PMA + IL-1β培养物中，没有一种抑制剂能保持的上调。TNF-α强烈介导PMA处理细胞中NFκB/p65的核转位，而IL-1β则不能。促炎细胞因子抑制PMA诱导的人THP-1细胞中的和mRNA。NFκB信号似乎是控制NOP转录的重要调节因子。这些发现表明，孤啡肽系统可能在免疫反应中发挥抗炎作用。
