Journal: International Journal of Biological Sciences

Article Title: Itaconate Modulates Neutrophil Homeostasis to Ameliorate Airway Inflammation in Diesel Exhaust Particles-exacerbated Asthma via Inhibiting NETs Formation

doi: 10.7150/ijbs.124927

Figure Lengend Snippet: DEP increases ITA production by inducing metabolic reprogramming in neutrophils. (A) Heatmap of differentially accumulated metabolites between control and HDM/DEP groups, and the change of ITA between control and HDM/DEP groups in the sequencing data. (B) Volcano plot of the differentially expressed genes between the control and HDM/DEP groups. (C) The levels of ITA measured by LC/MS in BALF and lung tissues of HDM/DEP-induced asthma (n = 6-8 mice per group). (D) Real-time qPCR analysis of Acod1 mRNA expression in the lung tissues of HDM/DEP-induced asthma (n = 4 mice per group). (E) Uniform manifold approximation and projection (UMAP) plot showing 13 clusters of cells in the lung tissues. (F) The percentage of the 13 clusters of cells in the lung tissues of con, HDM, and HDM/DEP groups. (G) The expression of Acod1 in different cell clusters. (H) Violin plot of the expression of Acod1 in different cell clusters. (I) Representative immunofluorescence staining of ACOD1 in the lung tissues of DEP-exposed asthmatic mice. Ly6G was used to mark neutrophils, and CD68 to mark macrophages. ACOD1 was co-localized with neutrophils. Scale bar, 20 μm. (J) Real-time qPCR analysis of Acod1 mRNA expression in BMDNs treated with DEP. (K) Representative immunofluorescence staining of ACOD1 in BMDNs treated with DEP. Scale bar, 10 μm. (L) Intracellular and extracellular levels of ITA measured by LC/MS in DEP-stimulated BMDNs. (M) Representative immunofluorescence staining of ACOD1 in cell pellets of BALF from healthy control and patients with severe asthma. Neutrophils were marked with CD16b, and macrophages were marked with CD68. ACOD1 was co-localized with neutrophils. Scale bar, 10 μm. (N) Real-time qPCR analysis of ACOD1 mRNA expression in PBNs stimulated with DEP. (O) Intracellular and extracellular levels of ITA measured by LC/MS in DEP-stimulated PBNs. (P) Integrated analysis of metabolomics and transcriptomics. (Q) Western blot analysis of TLRs and NF-κB signaling pathways in DEP-treated BMDNs. (R) Real-time qPCR analysis of Acod1 mRNA expression in DEP-stimulated BMDNs pre-treated with TLR2 inhibitor (C29), TLR4 inhibitor (TAK-242), and NF-κB inhibitor (BTZ). ** P < 0.01, compared with the control group; ## P < 0.01, ### P < 0.001, compared with the DEP group. (S) Western blot analysis of ACOD1 expression in DEP-stimulated BMDNs pre-treated with TLR2 inhibitor (C29), TLR4 inhibitor (TAK-242), and NF-κB inhibitor (BTZ). Data are presented as means ± SEM. ** P < 0.01, *** P < 0.001. HDM, house dust mite; DEP, diesel exhaust particles; LC/MS, Liquid Chromatography-Mass Spectrometry; BALF, bronchoalveolar lavage fluid; ACOD1, aconitate decarboxylase 1; BMDNs, bone marrow-derived neutrophils; PBNs, peripheral blood neutrophils; TLR, toll-like receptor; BTZ, Bortezomib.

Article Snippet: In some experiments, inhibitors targeting toll-like receptor 2 (TLR2) (C29, 100 μM; MedChemExpress, USA), TLR4 (TAK242, 100 nM; MedChemExpress, USA), the NF-κB signaling pathway (Bortezomib, 50 nM; MedChemExpress, USA), as well as ITA or 4-OI, were administered one hour prior to DEP or LPS treatment.

Techniques: Control, Sequencing, Liquid Chromatography with Mass Spectroscopy, Expressing, Immunofluorescence, Staining, Transcriptomics, Western Blot, Protein-Protein interactions, Liquid Chromatography, Mass Spectrometry, Derivative Assay

Journal: Neural Regeneration Research

Article Title: Overexpression of the inwardly rectifying potassium channel Kir4.1 or Kir4.1 Tyr 9 Asp in Müller cells exerts neuroprotective effects in an experimental glaucoma model

doi: 10.4103/NRR.NRR-D-24-00461

Figure Lengend Snippet: Kir4.1 overexpression and Kir4.1 Tyr 9 Asp overexpression in Müller cells suppress the MYD88/IRAK1/TRAF6/NF-κB p65 inflammatory signaling pathway. (A) Representative immunoblots showing changes in MYD88, IRAK1, TRAF6, and NF-κB p65 expression levels in microglia co-cultured with normal or activated Müller cells infected with LV-NC, Kir4.1, and Kir4.1 Tyr 9 Asp lentiviruses. (B, D, F, H) Bar charts summarizing the average densitometric quantification of immunoreactive bands for MYD88 (B), IRAK1 (D), TRAF6 (F), and NF-κB p65 (H) in microglia co-cultured with normal or activated Müller cells. n = 5 for each group. * P < 0.05, ** P < 0.01, vs. the + normal Müller cells group. Unpaired two-tailed t -test. (C, E, G, I) Bar charts summarizing the average densitometric quantification of immunoreactive bands for MYD88 (C), IRAK1 (E), TRAF6 (G), and NF-κB p65 (I) in microglia co-cultured with normal or activated Müller cells. n = 5 for each group. * P < 0.05, ** P < 0.01, vs. LV-NC + normal Müller cells group. One-way analysis of variance (Tukey–Kramer multiple comparisons test) was performed. eGFP: Enhanced green fluorescent protein; IRAK1: IL-1 receptor associated kinase 1; LV-NC: eGFP control lentiviruses; MYD88: myeloid differentiation primary response protein 88; NF-κB P65: nuclear factor kappa B P65; TRAF6: TNF receptor associated factor 6.

Article Snippet: Previous studies have demonstrated that Toll-like receptors play a role in microglial activation and the release of pro-inflammatory factors in the COH retina, a process mediated by the MYD88/IRAK1/TRAF6/NF-κB p65 signaling pathway (Takeda and Akira, 2004; Luo et al., 2010; Weber et al., 2010; Miao et al., 2023).

Techniques: Over Expression, Western Blot, Expressing, Cell Culture, Infection, Two Tailed Test, Control

Journal: Apoptosis

Article Title: Nicotine suppresses ferroptosis in colon cancer cells via HMOX1/NF-κB pathway to reduce oxaliplatin sensitivity

doi: 10.1007/s10495-026-02308-z

Figure Lengend Snippet: Nicotine reduces the sensitivity of colon cancer cells to oxaliplatin through modulation of the HMOX1/NF-κB signaling pathway. a-b The CCK-8 assay demonstrated that 10 µM oxaliplatin treatment for 24 h significantly inhibited the growth of LOVO and SW480 cells, while combined 1 µM nicotine treatment notably enhanced cell viability. Overexpression of HMOX1 reversed the 1 µM nicotine-induced increase in cell viability, and the 100 nM p-NF-κB pathway activator PMA treatment for 24 h partially restored this effect. c-d , k-l The scratch assay and quantitative analysis showed that HMOX1 overexpression reversed the 1 µM nicotine-induced increase in cell migration ability in LOVO and SW480 cells after 24 h of treatment, with 100 nM PMA treatment for 24 h partially restoring this effect. Scale bar = 20 μm. e-h The colony formation assay and quantitative analysis showed that HMOX1 overexpression reversed the 1 µM nicotine-induced enhancement of cell proliferation in LOVO and SW480 cells after 24 h, with 100 nM PMA treatment for 24 h partially restoring this effect. i-j , m-n The invasion assay and quantitative analysis showed that HMOX1 overexpression reversed the 1 µM nicotine-induced increase in cell invasion ability in LOVO and SW480 cells after 24 h of treatment, with 100 nM PMA treatment for 24 h partially restoring this effect. Scale bar = 20 μm. All data are expressed as mean ± standard deviation (independent triplicates). * p < 0.05; ** p < 0.01; *** p < 0.001; ns, no significant difference

Article Snippet: Iron death inducer RSL3 (Selleck, USA) and p-NF-κB signaling pathway activator PMA (Phorbol 12-myristate 13-acetate, Selleck, USA) were dissolved in DMSO to prepare stock solutions, stored at -20 °C in a dark environment, and diluted to working concentrations before use for cell treatment according to the experimental design.

Techniques: CCK-8 Assay, Over Expression, Wound Healing Assay, Migration, Colony Assay, Invasion Assay, Standard Deviation

Journal: International Dental Journal

Article Title: ARA290 Attenuates Apical Periodontitis via SIRT1/NF-κB/IL-1β Pathway Modulation

doi: 10.1016/j.identj.2025.100863

Figure Lengend Snippet: SIRT1/NF-κB/IL-1β signalling pathway was the potential target pathway for ARA290 to regulate the apical periodontitis course. A, Venn diagram of 17 intersecting target proteins from ARA290 and AP. B, The PPI network of intersecting target proteins from ARA290 and AP. C, KEGG pathway analysis of ARA290 in treating apical periodontitis. D, Immunohistochemical staining (scale bar = 50 μm) of SIRT1, acetylated NF-κB and IL-1β in the AP and ARA290 groups. E, The quantification of SIRT1, acetylated NF-κB and IL-1β (F) SIRT1-CD68 double labelling (scale bar = 50 μm) in the AP and ARA290 groups. Green, SIRT1; red, CD68; blue, DAPI. The red arrows represent cells with positive staining in the pathological staining figures; * P < .05, *** P < .005. AP, apical periodontitis; ARA290, apical periodontitis with the treatment of ARA290.

Article Snippet: Furthermore, to ascertain if ARA290 exerted its biological function via targeting the SIRT1/NF-κB signalling pathway during the progression of AP, ARA90 and the SIRT1 inhibitor selisistat (MCE) were used.

Techniques: Immunohistochemical staining, Staining

Journal: International Dental Journal

Article Title: ARA290 Attenuates Apical Periodontitis via SIRT1/NF-κB/IL-1β Pathway Modulation

doi: 10.1016/j.identj.2025.100863

Figure Lengend Snippet: Graphic abstract of the current research. A, In the progression of apical periodontitis, the pathogen-induced macrophage osteoclast differentiation via regulating SIRT1/NF-κB/IL-1β signalling pathway in Mф, which promoted the destruction of periapical bone tissues. B, The treatment of ARA290 could suppress the osteoclast differentiation via SIRT1/NF-κB/IL-1β in Mф, limiting the progression of apical periodontitis. The red arrow, activation; the green arrow, inhibition.

Article Snippet: Furthermore, to ascertain if ARA290 exerted its biological function via targeting the SIRT1/NF-κB signalling pathway during the progression of AP, ARA90 and the SIRT1 inhibitor selisistat (MCE) were used.

Techniques: Activation Assay, Inhibition