Fig 2: Schematic illustration demonstrates MyD88-dependent signaling pathways of neuropathic pain induced by CCI. Nerve injury produces abundant HMGB1and IL-1β in the DRG and SDH. The binding of HMGB1 and IL-1β to their receptors (TLR2/4 andIL-1R, respectively) activates MyD88 in the DRG and SDH, which phosphorylate NF-κB p65 and ERK. Phosphorylated NF-κB p65 subsequently enter the nucleus to regulate the expression of proinflammation cytokines such as TNF-α. Phosphorylated ERK enter the nucleus to induce transcription factors such as AP-1, which regulates the expression of certain cytokines and activates glial cells. All these signaling events consequently result in central and peripheral sensitizations that produce neuropathic pain

Fig 3: Wistin suppressed LPS-stimulated inflammation by inhibiting the phosphorylation of the p65 and p38 signaling pathways.

Fig 4: Inhibitory effects of CQMUH-011 on TLR4-MyD88 and NF-κB signaling. a Western blot analysis of TLR4 and MyD88 expression in LPS-induced primary microglial cells. Histograms presenting quantification of TLR4 and MyD88 expression normalized to endogenous β-actin in different groups. b Western blot analysis of P-p65, P-IκBα, and IκBα expression in LPS-induced primary microglial cells. Histograms presenting quantification of P-p65 expression normalized to endogenous β-actin and relative ratios of phosphorylated to total IκBα in different groups. Data are expressed as the means ± SD (n = 3 per group). *P < 0.05 LPS group vs. control group; #P < 0.05 CQMUH-011 group vs. LPS group.

Fig 5: Endometriosis upregulated neuronal pNF-κB-p65 in the dorsal root ganglion. a Western blot analyzed the time course of pNF-κB-p65 expression in the DRG from naïve, sham, 3 days, 7 days, 14 days, and 21 days after EM surgery. b Western blot revealed that EM upregulated pNF-κB-p65 protein expression in L3–L5 DRGs. N = 4, one-way ANOVA, *p < 0.05, versus sham and naïve. c–h Immunofluorescence showed neuronal distribution of pNF-κB-p65 in rat DRG of sham and EM 14 days groups. Scale bar 50 μm. i Immunofluorescence staining showed a significantly higher percentage of pNF-κB-p65-immunopositive DRG neurons in EM 14d group compared with that in sham group. n > 100 neurons from 3 rats in each group, *p < 0.05, EM 14d vs sham, chi-square test. j, k Western blot showed that HMGB1, TLR4, MyD88, and pNF-κB-p65 were upregulated in the ipsilateral DRG compared with the contralateral DRG. N = 4, *p < 0.05, Student’s t test, EM 14d-Ipsi versus EM 14d-Contra