Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Correlations between LL-37 and citLL-37-mediated increase in the mRNA abundance of COX-2 and chemokines. HBEC-3KT cells were stimulated with either (⬤) LL-37 or (🞅) citLL-37 (0.50 µM each). mRNA abundance of COX-2, IL-8, GROα and MIP-3α were examined in cell lysates using qRT-PCR after 4 h. Relative fold changes were calculated compared to unstimulated cells normalized to 1, using the ΔΔCt method after normalization with 18s RNA expression. Pearson’s correlation analysis was performed to determine the correlation between COX-2 and ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α mRNA abundance (fold changes compared to unstimulated cells)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, RNA Expression

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of COX-2 suppresses LL-37-mediated chemokine production. HBEC-3KT cells were pre-treated with COX-2 inhibitor Rofecoxib (20 nM) for 1 h, followed by stimulation with either LL-37, citLL-37 or sLL-37 (0.50 µM each) for 24 h (n = 5). TC supernatants were examined for the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α by ELISA. Chemokine levels shown in pg/mL after background subtraction of levels in unstimulated cells. Each dot represents an independent experiment (n = 5), the bars show IQR with the median line and the whiskers represent the min-max range. Statistical significance was measured using Two-Way ANOVA, and # represents statistical significance compared to unstimulated cells ( ** p < 0.005, *** p < 0.0005, #### or **** p < 0.0001)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of PGE2 receptors (EP1-4) suppresses LL-37-mediated enhancement of chemokines. HBEC-3KT cells were pre-treated with specific inhibitors for PGE2 receptors, EP1 (SC-19220; 20 nM), EP2 (PF-044EP2; 25 nM), EP3 (L-798,106; 20 nM), or EP4 (MF498; 10 nM), 1 h prior to stimulation with either LL-37or sLL-37 (0.50 μM). Tissue culture (TC) supernatants were collected after 24 h and the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α were measured by ELISA. Each dot represents an independent experiment (n=4), the bars show IQR with the median line and the whiskers represent the min-max range. Results shown are after subtracting baseline values obtained from unstimulated cells in each independent experiment. Statistical significance was determined using Two-Way ANOVA (** p < 0.001 and **** p < 0.0001)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Proposed mechanism of LL-37-COX-2 axis for chemokine production and neutrophil migration in the lungs. LL-37 engages the P2X 7 receptor in upregulating COX-2 expression which facilitates increase in the abundance of prostaglandin PGE2. Release of PGE2 may act in an autocrine manner through PGE2 receptors (EP1 − 4), resulting in enhanced production of chemokines including IL-8 and GROα, which facilitates neutrophil migration contributing to airway inflammation. However, citrullination of LL-37 dampens this pathway, potentially acting as a regulatory switch that limits the pro-inflammatory functions of LL-37 in the lungs (Figure created in BioRender.com)

Article Snippet: The abundance of IL-8 (Cat# DY208), GROα (Cat# DY275) and MIP-3α (Cat# DY360) were examined in TC supernatants using ELISA kits obtained from R&D Systems, according to the manufacturer’s instructions.

Techniques: Migration, Expressing

Journal: Pharmaceuticals

Article Title: Scutellarein from Erigeron breviscapus Inhibits Apoptosis-Mediated Epithelial Barrier Disruption and Alleviates Cigarette Smoke-Induced Lung Injury

doi: 10.3390/ph19010113

Figure Lengend Snippet: Scu reduces CS-induced inflammatory responses in BALF and lung tissue. Quantification of leukocytes ( A ), neutrophils ( B ), lymphocytes ( C ), and monocytes ( D ) in BALF from FA or CS-exposed mice receiving Veh (0.3% ( w / v ) CMC-Na solution containing 0.2% ( v / v ) Tween-80 and 0.2% ( v / v ) DMSO), 5, 10, 15 mg/kg Scu or 5 mg/kg Dex. Quantification of the contents of IL-1β ( E ), CXCL1 ( F ), TNF-α ( G ), and MCP-1 ( H ) in BALF from mice treated as in ( A – D ). Relative mRNA expression levels of Il1b ( I ), Cxcl1 ( J ), Tnf ( K ), Mcp-1 ( L ), and Mmp-9 ( M ) in lung tissue samples collected from differently treated mice. ( N ) Quantification of myeloperoxidase (MPO) in lung tissue samples collected from differently treated mice. Data represent mean ± SEM. N = 6; *, p < 0.05, **, p < 0.01, vs. FA-Veh; #, p < 0.05, ##, p < 0.01, vs. CS-Veh, by one-way ANOVA with Bonferroni’s multiple comparisons tests.

Article Snippet: ELISA kit of IL-1β (Cat# E-EL-M0037), TNF-α (Cat# E-EL-M3063), CXCL1 (Cat# E-EL-M0018), MCP1 (Cat# E-EL-M3001) were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Pharmaceuticals

Article Title: Scutellarein from Erigeron breviscapus Inhibits Apoptosis-Mediated Epithelial Barrier Disruption and Alleviates Cigarette Smoke-Induced Lung Injury

doi: 10.3390/ph19010113

Figure Lengend Snippet: Scu significantly alleviates CSC-induced pulmonary epithelial inflammation and barrier damage. Relative mRNA levels of Il1b ( A ), Cxcl1 ( B ), Tnf ( C ), Mcp-1 ( D ), and Mmp-9 ( E ) in MLE-12 cells treated with CSC for 12 h with or without Scu (1 μM) co-treatment. Data were expressed relative to the Veh. ( F ) Representative fluorescent images of MLE-12 cells stained with ZO-1, E-cadherin, Occludin, Claudin-18 (green), and Hoechst 33342 (blue) after being exposed to Veh (0.1% DMSO) or CSC (50 μg/mL) for 12 h with or without Scu (1 μM) co-treatment. Scale bar = 5 μm. ( G ) Changes in the TEER values of MLE-12 cell monolayer receiving Veh (0.1% DMSO) or CSC (50 μg/mL) treatment for 12 h with or without Scu (1 μM) co-treatment. ( H ) Changes in the Pd value of MLE-12 cells receiving Veh or CSC (50 μg/mL) for 12 h with or without Scu (1 μM) co-treatment. Data represent mean ± SEM. N = 6; **, p < 0.01, vs. Veh; #, p < 0.05, ##, p < 0.01, vs. CSC, by one-way ANOVA with Bonferroni’s multiple comparisons tests.

Article Snippet: ELISA kit of IL-1β (Cat# E-EL-M0037), TNF-α (Cat# E-EL-M3063), CXCL1 (Cat# E-EL-M0018), MCP1 (Cat# E-EL-M3001) were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Staining