雷米芬太尼诱导的小胶质细胞炎症:PAK4 介导的 NF-κB/NLRP3 通路的激活与痛觉减退的发生

IF 8.8 2区 医学 Q1 IMMUNOLOGY Brain, Behavior, and Immunity Pub Date : 2024-09-24 DOI:10.1016/j.bbi.2024.09.018
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引用次数: 0

摘要

背景围手术期使用瑞芬太尼会导致术后痛感亢进,从而影响术后恢复并延长住院时间。最近的研究发现,小胶质细胞介导的 NLRP3 炎性体的激活在阿片类药物诱导的痛觉减退中起着关键作用,NF-κB 是 NLRP3 的关键激活点。尽管有这些发现,但瑞芬太尼诱导术后痛觉减退的具体分子机制仍不清楚。本研究旨在建立瑞芬太尼诱导的痛觉减退模型,并采用体外和体内方法研究其分子机制,重点关注 NF-κB/NLRP3 通路。在体内经皮下注射 NLRP3 或 PAK4 拮抗剂,并使用 Von Frey 试验测量大鼠后爪的机械痛阈值。在体外,我们应用 NLRP3 或 PAK4 抑制剂或使用慢病毒感染来沉默 PAK4、NF-κB 和 NLRP3 基因。蛋白质表达通过免疫组化、免疫荧光和 Western 印迹进行评估。结果蛋白质组学分析表明,手术两小时后,瑞芬太尼可上调脊髓组织中的 PAK4 蛋白。此外,瑞芬太尼诱导脊髓背角发生形态学变化,其特征是 PAK4、p-p65、NLRP3 和 Iba-1 蛋白表达增加,进而导致 IL-1β 和 IL-18 水平升高和炎症反应。鞘内注射 NLRP3 或 PAK4 抑制剂可减轻瑞芬太尼引起的痛觉减退及相关变化。在体外,下调 PAK4 可抑制 LPS 诱导的 PAK4、p-p65、NLRP3 和 Caspase-1 的增加。结论瑞芬太尼可通过激活 NF-κB/NLRP3 通路和介导微神经胶质细胞活化,增加脊髓背角细胞中 PAK4 的表达,从而导致术后痛觉减退。
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Remifentanil-induced inflammation in microglial cells: Activation of the PAK4-mediated NF-κB/NLRP3 pathway and onset of hyperalgesia

Background

The perioperative use of remifentanil is associated with postoperative hyperalgesia, which can impair recovery and extend hospitalization. Recent studies have revealed that microglia-mediated activation of the NLRP3 inflammasome plays a critical role in opioid-induced hyperalgesia, with NF-κB acting as a pivotal activation point for NLRP3. Despite these findings, the specific molecular mechanisms underlying remifentanil-induced postoperative hyperalgesia remain unclear. This study aims to develop a model of remifentanil-induced hyperalgesia and investigate the molecular mechanisms, focusing on the NF-κB/NLRP3 pathway, using both in vitro and in vivo approaches.

Method

We established a remifentanil-induced hyperalgesia model and performed proteomic analysis to identify differential protein expression in the spinal cord tissue of rats. NLRP3 or PAK4 antagonists were administered intrathecally in vivo, and mechanical pain thresholds in the hind paws were measured using Von Frey testing. In vitro, we applied NLRP3 or PAK4 inhibitors or used lentivirus infection to silence PAK4, NF-κB, and NLRP3 genes. Protein expression was assessed through immunohistochemistry, immunofluorescence, and Western blotting. Additionally, ELISA was performed to measure IL-1β and IL-18 levels, and RT-qPCR was conducted to evaluate the transcription of target genes.

Results

Proteomic analysis revealed that remifentanil upregulates PAK4 protein in spinal cord tissue two hours after the surgery. In addition, remifentanil induces morphological changes in the spinal cord dorsal horn, characterized by increased expression of PAK4, p-p65, NLRP3 and Iba-1 proteins, which in turn leads to elevated IL-1β and IL-18 levels and an inflammatory response. Intrathecal injection of NLRP3 or PAK4 inhibitors mitigates remifentanil-induced hyperalgesia and associated changes. In vitro, downregulation of PAK4 inhibits the increase in PAK4, p-p65, NLRP3 and Caspase-1 induced by LPS. Conversely, the downregulation of NLRP3 does not impact the levels of PAK4 and p-p65 proteins, aligning with the in vivo results and suggesting that PAK4 acts as an upstream signaling molecule of NLRP3.

Conclusion

Remifentanil can increase PAK4 expression in spinal cord dorsal horn cells by activating the NF-κB/NLRP3 pathway and mediating microglial activation, thereby contributing to postoperative hyperalgesia.
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来源期刊
CiteScore
29.60
自引率
2.00%
发文量
290
审稿时长
28 days
期刊介绍: Established in 1987, Brain, Behavior, and Immunity proudly serves as the official journal of the Psychoneuroimmunology Research Society (PNIRS). This pioneering journal is dedicated to publishing peer-reviewed basic, experimental, and clinical studies that explore the intricate interactions among behavioral, neural, endocrine, and immune systems in both humans and animals. As an international and interdisciplinary platform, Brain, Behavior, and Immunity focuses on original research spanning neuroscience, immunology, integrative physiology, behavioral biology, psychiatry, psychology, and clinical medicine. The journal is inclusive of research conducted at various levels, including molecular, cellular, social, and whole organism perspectives. With a commitment to efficiency, the journal facilitates online submission and review, ensuring timely publication of experimental results. Manuscripts typically undergo peer review and are returned to authors within 30 days of submission. It's worth noting that Brain, Behavior, and Immunity, published eight times a year, does not impose submission fees or page charges, fostering an open and accessible platform for scientific discourse.
期刊最新文献
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