Fig 2: The relative expressions of proteins (A,B) and mRNAs (C) of the critical nodes in the TLR4/MyD88/NF-κB signaling pathways. Effects of TAK242 on TLR4, MyD88, TRAF6, P-IκBα and P-p65 expression stimulated by AMP-ZnONPs or not (D,E). The results were expressed as the mean ± SD (n = 3) (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. AMP-ZnONPs).

Fig 3: IRAK1 and TRAF6 are involved in expression of miR-146a-5p-suppressed inflammatory factors. TargetScan Human predicted (A) IRAK1 and (B) TRAF6 as target proteins of miR-146a-5p. Following transfection with miR-146a-5p mimics or inhibitor, Caco-2 cells were incubated with 10 ng/ml IL-1β for 24 h and collected. IRAK1 and TRAF6 protein was extracted, separated and analyzed by western blotting. (C) Relative protein expression of (D) IRAK1 and (E) TRAF6 in control, IL-1β, mimic NC, mimic NC + IL-1β, miR-146a-5p mimic and miR-146a-5p mimic + IL-1β groups were analyzed with Image J. (F) Relative protein expression of (G) IRAK1 and (H) TRAF6 in control, IL-1β, inhibitor NC, inhibitor NC + IL-1β, miR-146a-5p inhibitor and miR-146a-5p inhibitor + IL-1β groups were analyzed with Image J. *Ρ<0.05, **Ρ<0.01, ***Ρ<0.001 and ****Ρ<0.0001. IRAK1, interleukin 1 receptor associated kinase 1; TRAF6, tumor necrosis factor receptor-associated factor 6; UTR, untranslated region; miR, microRNA; NC, negative control.

Fig 4: Effect of LNT on the TLR4 signaling pathway in the mouse lung. The lungs of 5 dpi mice were collected, and the mRNA levels of TLR4 (A), MyD88 (B), TRAF6 (C) and NF-κB P65 (D) in the TLR4 signaling pathway were detected by RT–qPCR. LNT inhibited the expression levels of the four mRNAs. ###p < 0.001, vs control group (n=10); *p < 0.05, **p < 0.01, ***p < 0.001, vs. model group (n=10).

Fig 5: hucMSC-EV treat salpingitis by promoting macrophage polarization from M1 to M2 and inhibiting the TLR4 signaling pathway. hucMSC-EV downregulated M1 macrophages and cytokines TNF-α and IL-1β and upregulated M2 macrophages and cytokine IL-10. hucMSC-EV downregulated the TLR4, TRAF6, and p65 proteins to treat salpingitis and upregulated the pregnancy rate.