Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 2. ALOX5 expression is increased in RA CD4+ T cells. (A) Heatmap shows the expression of RA-related genes in peripheral blood CD4+ T cells from NCs and patients with RA. n = 8 NCs and 10 patients with RA. ALOX5 is highlighted with an arrow and red box. (B) Enrichment analysis for the top 10 diseases and functions represented by the transcripts showing differential expression in the NC and RA groups. Those in which ALOX5 was involved are outlined in red boxes. (C) ALOX5 transcript amounts in NC and RA CD4+ T cells as measured by RT-qPCR. n = 5 patients per group. (D) Representative immunoblot shows ALOX5 abundance in isolated CD4+ T cells from NCs and patients with RA. n = 5 patients per group. β-Actin is a loading control. (E) Quantification of ALOX5 protein in (D). Each protein quantification is based on data combined from three inde- pendent Western blots. n = 5 patients per group. (F) Representative immunoblot shows ALOX5 abundance in CD4+ T cells isolated from patients with RA treated with cortico- steroids (CS+, n = 3 patients) or untreated (CS−, n = 3 patients). ALOX5 protein abundance was quantified on the basis of data from three independent experiments. All data are presented as the mean ± SEM. Statistical significance was determined by Student’s t test; **P < 0.01.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques: Expressing, Quantitative Proteomics, Quantitative RT-PCR, Western Blot, Isolation, Control

Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 7. Association of ALOX5 with clinical remission in RA. (A) Immunoblotting for and quantification of ALOX5 abundance in NCs and patients with RA in dif- ferent phases of disease. β-Actin is a loading control. Each lane represents a single patient. Images are representative of n = 3 independent experiments with samples from different sets of patients. (B) Immunofluorescence staining for ALOX5 and CD3 in human synovial tissue from patients with high disease activity or in remission and quantification of the percentage of CD3+ cells that were also ALOX5+. n = 4 patients per group. (C) Calculated ALOX5 scores for 56 patients with RA with high disease activity or in remission. (D) ROC curve analyses were performed to assess the associations of high RA activity with several clinical parameters. The area under the curve values were 0.9572 for CRP (P < 0.0001), 0.8863 for the ESR (P < 0.0001), 0.7893 for the ALOX5 score (P = 0.0002), 0.6845 for RF (P = 0.0178), and 0.6501 for CCP (P = 0.0540). (E) Patients with RA in the remission and high disease activity groups were stratified into ALOX5high and ALOX5low groups (χ2, P = 0.0002). (F) Correlation matrix of the ALOX5 score and other clinical parameters in high-activity patients and remission patients. The power of the correlation is represented by circle color and size. For the patient data in (C) to (F), n = 27 patients in remission and 29 patients with high disease activity. All data are shown as the mean ± SEM. Statistical significances were determined by one-way ANOVA followed by Bonferroni’s multiple-comparisons test for multigroup comparisons or Student’s t test; *P < 0.05, **P < 0.01, and ****P < 0.0001.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Activity Assay

Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 8. Schematic model illustrating the proposed mechanism of CD4+ T cell pyroptosis in RA. In RA CD4+ T cells, ALOX5 is overexpressed, leading to an enhanced conversion of AA to LTB4, which subsequently increases ORAI3 abundance. ORAI3 oligomers form a Ca2+ channel that is activated by AA, and its increase results in increased Ca2+ influx into the cytoplasm. This surge in Ca2+ activates NLRP3, prompting it to form an inflammasome complex with the adapter protein ASC and pro- CASP1. This leads to autoprocessing of pro-CASP1 and the generation of active CASP1, which, in turn, cleaves GSDMD to yield GSDMD-N, inducing pyroptosis in CD4+ T cells.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: ACACA depletion activates NF-κB signaling in ARIPC cells. ( A ) Western blot analysis of key proteins of the NF-κB signaling pathway and inflammatory signaling in ACACA-depleted DU145 cells. ( B-C ) Western blot analysis of P65 and pP65 after treatment with TOFA for 72 h at the indicated concentrations (µg/mL) in the indicated ARIPC cells. ( D ) and ( E ) Western blot analysis of P65 and pP65 expression following ACACA silencing in the indicated ARIPC cells. ( F-I ) Immunofluorescence analysis of P65 in ACACA-depleted DU145 cells generated with different shRNA sequences (shACACA#1 and shACACA#2), with the quantification presented in ( G ) and ( I ), respectively. ( J-K ) Immunofluorescence analysis of P65 in DU145 cells after TOFA treatment (72 h), with quantification presented in ( K ). ( L ) Western blot analysis of P65 and pP65 and ( M ) qPCR analysis of P65 after arachidonic acid (AA) treatment (50 µM, 72 h). ( N ) Western blot analysis of P65 and pP65 and ( M ) qPCR analysis of P65 after cPLA2 silencing in ACACA-depleted DU145 cells. ( O ) qPCR analysis of P65. Data are presented as the means ± SD in ( G, I, K, M, O ). * P < 0.05

Article Snippet: The ACACA inhibitor TOFA (Abcam, #ab141578), the NF-κB inhibitor QNZ/EVP4593 (Selleckchem, #S4902), arachidonic acid (Selleckchem, #S6185), AR agonist R1881 ( GLPBIO, #GC19800), and AR antagonist Enzalutamide (Selleckchem, #S1250) were used at specified concentrations and time points as indicated.

Techniques: Western Blot, Expressing, Immunofluorescence, Generated, shRNA

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: Depletion of ACACA causes increased arachidonic acid activity in prostate cancer cells. ( A ) Levels of two metabolites associated with arachidonic acid from metabolomic analysis data after ACACA knockdown in DU145 cells. ( B-M ) Heatmap and individual panels of arachidonic acid and its metabolite levels in ACACA-depleted PC3 cells. ( N-O ) Heatmaps showing the transcriptional expression levels of arachidonic acid pathway metabolic enzymes in the indicated ARIPC cells. ( P-Q ) Indicated pathways significantly enriched in ACACA-depleted DU145 cells. Data are represented as mean ± SD in ( A , C-M ). * P < 0.05

Article Snippet: The ACACA inhibitor TOFA (Abcam, #ab141578), the NF-κB inhibitor QNZ/EVP4593 (Selleckchem, #S4902), arachidonic acid (Selleckchem, #S6185), AR agonist R1881 ( GLPBIO, #GC19800), and AR antagonist Enzalutamide (Selleckchem, #S1250) were used at specified concentrations and time points as indicated.

Techniques: Activity Assay, Knockdown, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: cPLA2-driven arachidonic acid (AA) promotes inflammation and migration in ACACA-depleted ARIPC cells. ( A ) qPCR analysis of inflammation biomarker-related gene expression in ACACA-depleted DU145 cells after 72 h of treatment with 50 µM AA. ( B-E ) Evaluation of the migration potential of ACACA-depleted DU145 cells following AA treatment (72 h) at the indicated concentrations, via wound healing ( B ) and Transwell assays ( D ), with the quantification presented in ( C ) and ( E ), respectively. ( F ) Transcriptome data and ( G ) Western blot analysis of cPLA2 expression in ACACA-depleted DU145 cells. ( H ) Correlation analysis between cPLA2 and ACACA expression levels in the SU2C PolyA cohort. ( I ) Violin plots illustrating cPLA2 expression levels in mCRPC samples divided into high and low ACACA expression groups in the SU2C PolyA cohort. ( J ) qPCR analysis of cPLA2 expression after 72 h of treatment with silencing RNA in ACACA-depleted DU145 cells. ( K ) AA concentration measured by ELISA following cPLA2 silencing for 72 h in ACACA-depleted DU145 cells. ( L ) qPCR analysis of inflammation biomarker-related gene expression after cPLA2 silencing in ACACA-depleted DU145 cells. ( M-P ) Evaluation of the migration potential of ACACA-depleted ARIPC cells after cPLA2 silencing via wound healing ( M ) and Transwell ( O ) assays, with the quantification presented in ( N ) and ( P ), respectively. The data are presented as the means ± SDs in A, C, E, F, I, J-L, N , and P. * P < 0.05

Article Snippet: The ACACA inhibitor TOFA (Abcam, #ab141578), the NF-κB inhibitor QNZ/EVP4593 (Selleckchem, #S4902), arachidonic acid (Selleckchem, #S6185), AR agonist R1881 ( GLPBIO, #GC19800), and AR antagonist Enzalutamide (Selleckchem, #S1250) were used at specified concentrations and time points as indicated.

Techniques: Migration, Biomarker Discovery, Gene Expression, Western Blot, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: A Metabolomics Study on the Bone Protective Effects of a Lignan-Rich Fraction From Sambucus Williamsii Ramulus in Aged Rats

doi: 10.3389/fphar.2018.00932

Figure Lengend Snippet: Identified differential metabolites in serum accountable for the discrimination between OVX rats and Sham rats by UPLC-MS in negative ESI mode.

Article Snippet: Taurocholic acid was obtained from Steraloids (Newport, RI, USA). p -Cresyl sulfate potassium salt and arachidonic acid were supplied from Toronto Research Chemicals (North York, ON, Canada).

Techniques:

Journal: Advanced Science

Article Title: The Long Non‐Coding RNA Obesity‐Related (Obr) Contributes To Lipid Metabolism Through Epigenetic Regulation

doi: 10.1002/advs.202401939

Figure Lengend Snippet: Obr is a target of Pparγ, and the HFD diet induces Obr expression through Pparγ. A,B) C9 cells were treated with two different concentrations of Pparγ antagonist (Sw‐9662) or agonist Troglitazone, and the expression of Pparγ and Obr was analyzed by qRT‐RCR. The expression of Obr decreased (A) or increased (B) in response to changes in Pparγ expression. C) The Pparγ response elements (PREs) regions were tested in the Obr promoter. D,E) C9 liver cells were treated with Pparγ antagonist (Sw‐9662) or the agonist Troglitazone, and the binding of Pparγ to two different PRE‐regions of the Obr promoter was analyzed by ChIP analysis. The binding decreased in the presence of the antagonist (D) and increased with the activation of Pparγ (E). F) C9 cells were treated with two different concentrations of polyunsaturated lipid, Arachidonic acid, which is present in HFD, and the expression of Pparγ and Obr was analyzed by qRT‐RCR. Both the expression of Obr and Pparγ were increased in response to the addition of Arachidonic acid. G) C9 cells were treated with polyunsaturated lipid, Arachidonic acid as above, and the binding of Pparγ to two different PREs mentioned above was tested by ChIP analysis. The binding increased in the presence of Arachidonic acid. H) Using liver tissue samples from normal chow and HFD‐fed animals, we have also verified the binding of Pparγ to the PREs of the Obr promoter mentioned above by ChIP analysis. The binding of Paprγ to the Obr promoter was increased in the livers of HFD‐fed animals. I) The model depicts the association of Obr with the CREB histone acetyltransferase complex. An increase in the lipid levels from the HFD or other sources increased the expression of Pparγ, which increased the expression of Obr. Obr associated with the CREB histone acetyltransferase complex, and this association promoted H3K27 acetylation at promoters of H3K27 acetylation at promoters of the target genes. Data are given as mean ± SE. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: C9 cells were cultured in 6 well plates for 24 h before treating the cells with 10 and 100 μ m GW9662 (MedChem Express), 100, and 150 μ m Troglistazone (Sigma) or 50 and 100 μ m arachidonic acid (Santa Cruz).

Techniques: Expressing, Binding Assay, Activation Assay