Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: H1N1 infection affects cell viability, inflammatory cytokine secretion and interactions between HBEpiCs and THP-1 cells. (A) CCK-8 assay revealed that HBEpiC viability decreased in a concentration-dependent manner following H1N1 infection. (B) ELISA revealed that the levels of IL-1β, IL-6, TNF-α, and IL-8 in HBEpiCs decreased with increasing H1N1 infection. (C) CCK-8 assay indicated that supernatants from H1N1-infected HBEpiC cultures reduced the viability of THP-1 cells in a dose-dependent manner. (D) ELISA results suggested that the levels of inflammatory cytokines (IL-1β, IL-6, TNF-α and IL-8) in THP-1 cells were decreased following exposure to supernatants from H1N1-infected HBEpiC cultures. (E) Cell adhesion assay revealed that the number of THP-1 cells adhering to HBEpiCs increased with increasing H1N1 concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) Transwell assay suggested that H1N1 infection enhanced the migration capacity of THP-1 cells, with increased migration observed at higher virus concentrations (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; ** P<0.01, *** P<0.001 vs. control. H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control; MOI, multiplicity of infection.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Infection, CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Adhesion Assay, Transwell Assay, Migration, Virus, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: TP modulates the inflammatory response and immune cell activity in H1N1-infected HBEpiCs and THP-1 cells. (A) No significant changes were observed in HBEpiCs treated with various concentrations of TP (5, 10 and 20 nM) following H1N1 infection compared with the control. (B) After TP treatment, the levels of the inflammatory cytokines IL-1β, IL-6, TNF-α and IL-8 in HBEpiCs were markedly lower than those in the untreated group. (C) The viability of THP-1 cells pretreated with H1N1-infected HBEpiC culture supernatant decreased after TP treatment. (D) The levels of IL-1β, IL-6, TNF-α, and IL-8 in THP-1 cells were markedly lower after TP treatment. (E) The adhesion of THP-1 cells to HBEpiCs induced by H1N1 infection decreased in a dose-dependent manner with increasing TP concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) The migration capacity of THP-1 cells was markedly reduced when the supernatant from H1N1-infected HBEpiC cultures was treated with TP (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. control. TP, triptolide; H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Activity Assay, Infection, Control, Concentration Assay, Migration, Standard Deviation

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation increases expression of neutrophil recruitment and NF-kB signaling genes by BM-hMSCs. Heatmap clustering of the Z-score of the top 20 differentially expressed genes between unstimulated control BM-hMSCs (3 replicates, 1 donor, green) and IL-1 β stimulated BM-hMSCs (3 replicates, 1 donor, purple). Red color indicates higher expression of the genes and blue color indicates decreased expression A . Protein expression of CXCL1 (10 replicates, 4 donors), CCL2 (4 replicates, 2 donors), CXCL5 (4 replicates, 2 donors), and CXCL8/IL-8 (7 replicates, 3 donors) in BM-hMSCs secretome measured by ELLA or ELISA. Data are presented as median, and statistical analysis was performed using unpaired t-test with Welch’s correction B . Bar plot of the top 20 significant Gene Ontology (GO) biological processes representing IL- 1β induced upregulated differentially expressed genes C IL-1 β, Interleukin-1β; BM-hMSCs, Bone marrow derived human mesenchymal cells; CCL2, chemokine (C–C motif) ligand 2; CXCL1, chemokine (C-X-C motif) ligand 1; CXCL2, chemokine (C-X-C motif) ligand 1; CXCL8, chemokine (C-X-C motif) ligand 8; ****, p < 0,0001

Article Snippet: All samples were diluted 1:1 and the analytes were measured using custom made cartridges that included CCL2, CXCL5, and CXCL8 (Bio-techne) or measured by Quantikine ELISA (cat# D8000C, R&D Systems) with 1:1 dilution.

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) Cell viability was assessed with the MTT assay and data shown are means ± SD (n = 3). ( b ) TNFα, IL-6 and IL-8 expression in response to B[a]P exposure. ( c ) Representative Western blot images showing protein expression. All data were obtained from three independent experiments. Statistical p values were determined by a one-way ANOVA, followed by a Tukey post-hoc test comparing the B[a]P treatments to the DMSO control; * p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: Cells were treated with various concentrations of PM2.5 for 24 hr. ( a ) Cell viability was assessed with MTT assay. ( b ) IL-6 and IL-8 from the basolateral sides. ( c ) Representative Western blot images showing CFTR, β2AR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), NF-κB p65, and TRPC6 expression. All data are presented as the mean ± SD (n = 4). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; *, p < 0.05; ¥, p < 0.01; NS, no significance.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: MTT Assay, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ALI cells were treated with DMSO, 100 ng/mL PM 2.5, 10 µM LY 294002 ± PM 2.5, or 10 µM LY 303511 ± PM 2.5 for 24 hrs. ( a ) IL-6 and IL-8 from the basolateral sides were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n=3). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing all treatments to the DMSO control, or PM 2.5 to PM 2.5 ± LY 294002 or LY 303511; *, p < 0.01. ( b ) Representative Western blot images showing CFTR and β-actin expression. β-actin was used to ensure equal loading of protein. All data were obtained from three independent experiments, and each experiment was analyzed in duplication.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

doi: 10.64898/2026.04.21.719931

Figure Lengend Snippet: ( a ) The lung parenchyma from two ferrets were sliced into several sections as shown. Arrows indicate the tracheobronchial tubes. All tissue sections were incubating with DMSO (control), 10 µM B[a]P, or 100 ng/mL PM 2.5 in Pneumacult-EX Plus Basal media for 24 hrs. ( b ) IL 6 secretion was determined using the ferret ELISA kit. Data represent the mean of 6 independent sections. Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; ¥, p < 0.05 (n = 6). ( c ) Representative western blotting images show reduction of CFTR, and β2AR and elevation of TRPC6 expression in explants under these treatments.

Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Immunity, Inflammation and Disease

Article Title: Ginkgolide B Alleviates Airway Inflammation in Hyperoxia Lung Injury

doi: 10.1002/iid3.70364

Figure Lengend Snippet: PAF stimulates neutrophils to be resident locally and secrete IL‐6. (A) The level of PAF in plasma from preterm neonates with BPD ( n = 8) was higher than that in non‐BPD group ( n = 8). (B) The level of PAF in BALF from preterm neonates with BPD ( n = 14) increased higher than that in non‐BPD group ( n = 9). (C) PAF level in BALF was positively correlated with oxygen supplement days in patients with BPD ( n = 14). (D–F) The percentages of CD66b + CD62L + in neutrophils from the blood of healthy neonates at birth decreased sharply after PAF (100 ng/mL) stimulation at different time points. (G) The level of soluble CD62L in the plasma of patients with BPD ( n = 11) or without BPD ( n = 15). (H–K) Relative mRNA expressions of IL6 , LCN2 , IL10 , and MMP12 were examined in neutrophils after PAF (100 ng/mL) stimulation for 4 h. (L) PAF induced neutrophils to form NETs (MPO represents neutrophil activation, and the red arrows indicate the presence of free DNA outside the cells, which signifies the formation of NETs). (M) Clustering of inflammatory cytokine levels in BALF from preterm neonates with and without BPD. (N) Differential inflammatory cytokines in BALF from patients with and without BPD were displayed in heatmap. (O–R) The levels of IL‐6, IL‐8, and GM‐CSF were tested higher in BALF of BPD patients ( n = 9–26) postnatal 1 week than those of non‐BPD ( n = 6–18), except for GRO‐α by ELISA. Each experiment was independently repeated at least three times.

Article Snippet: Human IL‐8 Precoated ELISA Kit , DAKEWE , 1110802 , .

Techniques: Clinical Proteomics, Activation Assay, Enzyme-linked Immunosorbent Assay