DEAD-box helicase 54 regulates microglial inflammatory response in rats with chronic constriction injuries through NF-κB/NLRP3 signaling axis.

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-1ß, 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-1ß, 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.

Resolvin D1 attenuates mechanical allodynia after burn injury: Involvement of spinal glia, p38 mitogen-activated protein kinase, and brain-derived neurotrophic factor/tropomyosin-related kinase B signaling.

Resolvin D1 (RvD1) suppresses inflammatory, postoperative, and neuropathic pain. The present study assessed the roles and mechanisms of RvD1 in mechanical allodynia after burn injury. A rat model of burn injury was established for analyses, and RvD1 was injected intraperitoneally. Pain behavior and the expression levels of spinal dorsal horn Iba-1 (microglia marker), GFAP (astrocyte marker), p-p38 mitogen-activated protein kinase (MAPK), brain-derived neurotrophic factor (BDNF), and tropomyosin-related kinase B (TrkB) were detected by behavioral and immunocytochemical assays. The results showed that RvD1 attenuated mechanical allodynia after burn injury, prevented microglial and astroglial activation, and downregulated p-p38 MAPK in microglia and BDNF/TrkB following burn injury. Similarly, inhibition of p38 MAPK and BDNF/TrkB signaling attenuated mechanical allodynia after burn injury. In addition, inhibition of p38 MAPK prevented spinal microglial activation and downregulated BDNF/TrkB following burn injury. Furthermore, inhibition of BDNF/TrkB signaling prevented spinal microglial activation and downregulated p-p38 MAPK within spinal microglia. Taken together, this study demonstrated that RvD1 might attenuate mechanical allodynia after burn injury by inhibiting spinal cord glial activation, microglial p38 MAPK, and BDNF/TrkB signaling in the spinal dorsal horn.

Highly sensitive non-enzymatic hydrogen peroxide monitoring platform based on nanoporous gold via a modified solid-phase reaction method.

In this work, nanoporous gold (NPG) fabricated using a modified solid-phase reaction method was developed as an electrocatalyst for the nonenzymatic detection of hydrogen peroxide (H2O2). The NPG morphology and structure were characterized by scanning electron microscopy and high-resolution transmission electron microscopy. The fabricated NPG exhibited a nanoporous framework with numerous structural defects. The NPG-based amperometric H2O2 sensor had a good selectivity, reproducibility, and low detection limit (0.3 µM) under near physiological conditions (pH = 7.4). The sensitivities of this sensor over concentration ranges of 0.002-5 mM and 5-37.5 mM were 159 and 64 µA mM-1 cm-2, respectively. These results indicate that the developed NPG is a promising material for the electrochemical sensing of H2O2.

The Roles of Chemokine CXCL13 in the Development of Bone Cancer Pain and the Regulation of Morphine Analgesia in Rats.

Chemokines are important regulators of immune, inflammatory, and neuronal responses in peripheral and central pain pathway. The aim of this study was to investigate whether chemokine (C-X-C motif) ligand 13 (CXCL13) and its receptor (C-X-C chemokine receptor type 5, CXCR5) involve in the development of bone cancer pain (BCP) and the regulation of morphine analgesia in rats. The change of pain behaviors in BCP rats were measured by testing paw withdrawal threshold (PWT). The levels of CXCL13, CXCR5 and signal pathway proteins (p-p38, p-ERK and p-AKT etc.) in the spinal cord were measured via western blots. The expression of CXCL13 and CXCR5 in spinal cord was increased in BCP rats. The BCP rats showed decrease of PWTs, which was relieved by CXCR5i. Intrathecally injection of murine recombinant CXCL13 (mrCXCL13) decreased the PWTs of BCP rats and opposed morphine-induced analgesia in BCP rats. In BCP rats, the signal pathway proteins (p38, ERK and AKT) in the spinal cord were activated. CXCL13 and morphine had contrary effect on the phosphorylation of these proteins. MrCXCL13 directly increased the levels of p-p38, p-ERK and p-AKT in BCP rats. However, morphine decreased the levels of these proteins in BCP rats. While blocking the activation of p-p38, p-ERK and p-AKT, morphine analgesia was enhanced. These results suggest CXCL13 participated in bone cancer pain and opposed morphine analgesia via p38, ERK and AKT pathways. It may be a target to enhance pain management in cancer pain patients.

[Resolvin D1 inhibits the injury of PC12 cells induced by activated microglia].

Objective To investigate the effect of resolvin D1 (RvD1) on the injury of PC12 cells induced by activated BV-2 microglia and the related mechanisms. Methods BV-2 cells were divided into control group, lipopolysaccharide (LPS)-treated group, RvD1-treated group and RvD1 combined with LPS (RvD1-LPS)-treated group. After BV-2 cells were incubated with the corresponding substances for 12 and 24 hours, the levels of interleukin-1ß (IL-1ß), IL-6, tumor necrosis factor-α (TNF-α) in the supernatants were determined by ELISA. The culture supernatants of BV-2 cells were collected at 24 hours and added into PC12 cells for another 24-hour culture. Thereafter, the survival rate of PC12 cells was tested by MTT assay. The expression of NF-κB p65 protein in BV-2 cells was deteced by Western blotting. Results Compared with the control group, the survival rate of PC12 cells in the LPS group significantly decreased; the levels of IL-1ß, IL-6 and TNF-α in the supernatant of BV-2 cells and the nuclear translocation of NF-κB p65 significantly increased in the LPS group. Compared with the LPS group, the survival rate of PC12 cells in RvD1-LPS group was significantly elevated; the levels of IL-1, IL-6, TNF-α and the nuclear translocation of NF-κB p65 were significantly reduced in RvD1-LPS group. Conclusion RvD1 can inhibit the injury of PC12 cells induced by activated BV-2 microglia through inhibiting the nuclear translocation of NF-κB p65 and inflammatory factor levels in BV-2 cells.