mouse il 1β antisera  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: The Relationship Between Decreased Serum Klotho Protein and Cognitive Impairment in Patients After Abdominal Surgery

doi: 10.2147/JIR.S593405

Figure Lengend Snippet: The correlation between serum Klotho and clinical indicators. Pearson correlation test was performed between Klotho with ( A ) duration of anesthesia, ( B ) postoperative MoCA scores, ( C ) postoperative CRP, ( D ) postoperative IL-1β, and ( E ) postoperative IL-6 in all 93 cases of patients after abdominal surgery.

Article Snippet: CRP (catalog no. SEKH-0138) and Klotho (catalog no. SEKH-0280) ELISA kits were purchased from Solarbio (Beijing, China), while IL-1β (catalog no. DLB50) and IL-6 (catalog no. D6050B) ELISA kits were obtained from R&D Systems (Minneapolis).

Techniques:

Journal: Journal of Inflammation Research

Article Title: The Relationship Between Decreased Serum Klotho Protein and Cognitive Impairment in Patients After Abdominal Surgery

doi: 10.2147/JIR.S593405

Figure Lengend Snippet: Proposed mechanism linking decreased postoperative serum Klotho levels to postoperative cognitive dysfunction (POCD) after abdominal surgery. Surgery and prolonged anesthesia lead to a reduction in serum Klotho levels, which contributes to increased oxidative stress and systemic inflammation, as indicated by elevated levels of CRP, IL-1β, and IL-6. These processes promote neuroinflammation and synaptic dysfunction, resulting in cognitive decline and POCD. Arrows indicate the direction of the effects, illustrating how decreased Klotho mediates the relationship between perioperative stress, inflammation, and postoperative cognitive impairment.

Article Snippet: CRP (catalog no. SEKH-0138) and Klotho (catalog no. SEKH-0280) ELISA kits were purchased from Solarbio (Beijing, China), while IL-1β (catalog no. DLB50) and IL-6 (catalog no. D6050B) ELISA kits were obtained from R&D Systems (Minneapolis).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits activation of the NLRP3 inflammasome. (A) Schematic workflow of in vitro NLRP3 inflammasome activation. (B,C) Cell viability was assessed in J774A.1 cells (B) and PMA (500 nM)-differentiated THP-1 cells (C) treated with TMH for 2 or 18 h using the EZ-Cytox assay. (D) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with ATP (5 mM) for 30 min in the presence or absence of MCC950. IL-1β (p17) levels in supernatants were measured by ELISA. (E,F) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with ATP (5 mM) (E) or nigericin (10 μM) (F) for 1 h, with or without MCC950. IL-1β (p17) levels in supernatants were determined by ELISA. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits activation of the NLRP3 inflammasome. (A) Schematic workflow of in vitro NLRP3 inflammasome activation. (B,C) Cell viability was assessed in J774A.1 cells (B) and PMA (500 nM)-differentiated THP-1 cells (C) treated with TMH for 2 or 18 h using the EZ-Cytox assay. (D) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with ATP (5 mM) for 30 min in the presence or absence of MCC950. IL-1β (p17) levels in supernatants were measured by ELISA. (E,F) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with ATP (5 mM) (E) or nigericin (10 μM) (F) for 1 h, with or without MCC950. IL-1β (p17) levels in supernatants were determined by ELISA. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH does not affect other inflammasome or inflammatory pathways. (A,B) LPS-primed J774A.1 cells were treated with TMH for 2 h, followed by stimulation with either dsDNA (1 μg/mL) (A) or flagellin (B) transfected using Lipofectamine 2000 for 3 h. (C) Following TMH treatment (2 h), nuclear and cytosolic fractions of LPS-primed J774A.1 cells were separated and assessed for NF-κB p65 translocation by immunoblotting. (D) HEK293FT cells transfected with the pGL4.32 luciferase reporter vector were treated with TNF-α (20 ng/mL) for 5 h, with or without TMH pre-treatment (2 h). Luciferase activity was measured using the Bright-Glo™ luciferase assay system (Promega). Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Transfection, Translocation Assay, Western Blot, Luciferase, Plasmid Preparation, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits NLRP3 inflammasome activation by suppressing ASC oligomerization and ATPase activity. (A) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with imiquimod (200 μM) for 1 h. (B) Cells were treated with TMH (2 h), stained with MitoSOX (5 μM, 10 min), and stimulated with ATP (5 mM, 5 min). Intracellular ROS were visualized by confocal microscopy. (C,D) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with nigericin (10 μM, 1 h). ASC speck formation was examined by confocal microscopy (C) , and representative images from five fields are shown (D) . (E) Cells were stimulated with or without MCC950 and cross-linked with DSS (2.5 mM, 30 min) before immunoblotting for ASC oligomerization. (F) NLRP3 ATPase activity was measured using the ADP-Glo™ assay to quantify ATP-to-ADP conversion following TMH treatment. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, Activity Assay, Staining, Confocal Microscopy, Western Blot, Glo Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: Toxicity assessment of TMH in zebrafish embryos. (A) Survival curves of zebrafish embryos treated with TMH at concentrations of 1, 10, or 100 μg/mL. Sixty embryos were used per condition and maintained in E3 medium. The control group was treated with DMSO (0.25%, v/v), corresponding to the highest TMH concentration. (B) Survival and phenotype distribution of 5 dpf embryos treated with TMH at distinct concentrations from 1 to 5 dpf. (C) Representative images showing developmental abnormalities observed in embryos treated with 10 μg/mL TMH at 5 dpf. (D) Representative images of Sudan Black B staining in 3 dpf embryos treated with vehicle control (0.0025% DMSO with PTU; upper panel) or TMH (1 μg/mL; lower panel). (E) Quantification of neutrophil counts in the CHT (red dashed outline) following treatment with TMH (1 μg/mL). Data are presented as mean ± SEM of individual embryos. Statistical significance was determined using an unpaired two-tailed Student’s t-test; n.s., not significant (p = 0.9997). Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Control, Concentration Assay, Staining, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH suppresses LPS-induced inflammatory cell recruitment in zebrafish embryos. (A) Representative images of neutrophil recruitment in the CHT in 3 dpf zebrafish embryos following LPS stimulation, assessed by Sudan Black B staining. Top panel: control embryos treated with 0.0025% DMSO (n = 29); middle panel: LPS-treated embryos (10 μg/mL; n = 32); bottom panel: embryos co-treated with LPS (10 μg/mL) and TMH (1 μg/mL; n = 33). (B) Quantification of neutrophil accumulation in the CHT following LPS stimulation with or without TMH treatment using Sudan Black B staining from (A) . (C) Representative images of whole-mount in situ hybridization showing mpx -positive neutrophils in 3 dpf embryos under LPS and TMH treatment conditions. Top panel (DMSO 0.0025%; n = 34); middle panel (LPS 10 μg/mL; n = 35); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 39). (D) Quantification of mpx -positive cells within the CHT from (C) . (E) Representative confocal images of macrophages in 3 dpf Tg ( mpeg1 :EGFP) zebrafish embryos following LPS and TMH treatment. Top panel (DMSO 0.0025%; n = 8); middle panel (LPS 10 μg/mL; n = 11); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 13). (F) Quantification of mpeg1 :EGFP-positive macrophages in the CHT region shown in (E) . Red dashed outline indicates the CHT. All data are presented as mean ± SEM. Statistical significance was determined using an unpaired two-tailed Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.05; ****P < 0.0001. Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Staining, Control, In Situ Hybridization, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Secretory VSMC-derived THBS1 promotes macrophage M1 polarization and inflammation in intracranial aneurysms via the CD47/NF-κB axis

doi: 10.3389/fimmu.2026.1790488

Figure Lengend Snippet: Secretory VSMCs promote macrophage M1 polarization and inflammatory activation through the THBS1/CD47/NF-κB axis. (A) ELISA quantification of THBS1 secretion from HA-VSMCs before and after TGF-β1/PDGF-BB stimulation and following shTHBS1 transduction. (B–D) qRT-PCR analysis of macrophage IL-1β, TNF-α, and iNOS mRNA in the Transwell co-culture system. (E, F) ELISA quantification of IL-1β and TNF-α secretion in co-culture supernatants. (G, H) Flow cytometric analysis of CD86 + macrophages and NF-κB transcriptional activity (Dual-luciferase reporter assay). All in vitro experiments were independently performed in three biological replicates (n = 3). Data are presented as mean ± standard deviation (SD). Statistical comparisons among multiple groups were performed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple-comparison test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: At the protein secretion level, commercially available Quantikine ® ELISA kits (R&D Systems, DY201 for IL-1β; DY210 for TNF-α) were used to quantify IL-1β and TNF-α concentrations in the lower-chamber supernatants following the manufacturer’s instructions.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Transduction, Quantitative RT-PCR, Co-Culture Assay, Activity Assay, Luciferase, Reporter Assay, In Vitro, Standard Deviation, Comparison