Abstract：
Neuropathic pain (NP) is caused by damage to or disease of the somatosensory nervous system, but its mechanism is still not fully understood. In this study, DEAD-box helicase 54 (DDX54) was targeted, and its regulatory role was explored in a chronic constriction injury (CCI) rat model. Microglia and HMC3 cells were stimulated with LPS. The interaction between DDX54 and myeloid differentiation factor-88 adapter protein (MYD88) was verified. A CCI of sciatic nerve model in rats was established. Behavioral testing was performed before and after the CCI. The expressions of IL-1β, TNF-α, and IL-6 were upregulated, and those of DDX54, MYD88, NF-κB, and NOD-like receptor 3 (NLRP3) were upregulated in microglia and HMC3 cells after LPS induction. DDX54 knockdown in microglia and HMC3 cells inhibited IL-1β, TNF-α, and IL-6 expressions and downregulated the protein levels of MYD88, p-NF-κB p65 (p-p65), and NLRP3. DDX54 overexpression promoted the stability of MYD88 mRNA. DDX54 binds to the MYD88-3\'-untranslated region (UTR). DDX54 interference in rats could alleviate the decrease of paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) induced by CCI, inhibit Iba1 expression, and reduce inflammatory factors as well as MYD88 and NF-κB expressions. DDX54 promotes the activation of NF-κB/NLRP3 signaling by regulating MYD88 mRNA stability, thereby affecting inflammatory response and NP progression in CCI rats.NEW & NOTEWORTHY The role of DDX54 protein in LPS-induced microglia and a chronic constriction injury (CCI) rat model was investigated for the first time. DDX54 interference can inhibit microglial activation and reduce the secretion of inflammatory factors. The interaction between DDX54 protein and MYD88 mRNA was explored for the first time. DDX54 promotes NF-κB/NLRP3 signaling activation by regulating MYD88 transcription in a CCI rat model.
摘要：
目的神经性疼痛（NP）是由体感神经系统的损害或疾病引起的，但其机制仍未完全理解。在这项研究中,DEAD-box解旋酶54(DDX54)被靶向,并在慢性缩窄性损伤（CCI）大鼠模型中探讨了其调节作用。材料和方法用LPS刺激小胶质细胞和HMC3细胞。验证了DDX54与髓样分化因子88衔接蛋白（MYD88）之间的相互作用。建立大鼠坐骨神经CCI模型。在CCI之前和之后进行行为测试。结果IL-1β的表达,TNFα和IL-6上调，LPS诱导后，小胶质细胞和HMC3细胞中DDX54，MYD88，NF-κB和NLRP3的表达上调。DDX54敲低小胶质细胞和HMC3细胞抑制IL-1β，TNFα和IL-6表达，下调MYD88、p-p65和NLRP3的蛋白水平。DDX54过表达促进了MYD88mRNA的稳定性。DDX54与MYD88-3UTR区结合。DDX54干预大鼠可减轻CCI引起的PWMT和PWTL的降低，抑制Iba1表达，降低炎症因子以及MYD88和NF-κB的表达。结论DDX54通过调节MYD88mRNA的稳定性促进NF-κB/NLRP3信号的激活,从而影响CCI大鼠的炎症反应和NP进展。
