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 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