Journal: bioRxiv

Article Title: OLT1177 (Dapansutrile) inhibits Gasdermin D-dependent IL-1β Release and Pyroptotic Cell Death in Bone Marrow-derived Macrophages

doi: 10.64898/2026.05.31.729061

Figure Lengend Snippet: BMDMs from WT mice were treated with either media alone, lipopolysaccharide, LPS alone, LPS plus nigericin (LPS + NIG), or LPS + NIG plus OLT1177 (NLRP3 inhibitor). Nigericin (10 μM) was added to BMDMs after 3.5 hours of stimulation and cytokines were measured in the culture supernatants at 4h. IL-1β (A), CCL3 (B), and MPO (C) were measured in the culture supernatants by the R&D Duo set ELISA kit. Data are presented as mean ± SEM from three independent biological replicates (BMDMs derived from three independent mice). ****P< .0001.

Article Snippet: After 3.5 hours, the NLRP3 inflammasome formation was induced with 10 μM nigericin (InvivoGen, San Diego, CA) for 30 minutes.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: bioRxiv

Article Title: OLT1177 (Dapansutrile) inhibits Gasdermin D-dependent IL-1β Release and Pyroptotic Cell Death in Bone Marrow-derived Macrophages

doi: 10.64898/2026.05.31.729061

Figure Lengend Snippet: A) BMDMs were treated with lipopolysaccharide, LPS (1μg/mL) and LPS plus nigercin (NIG). Representative western blots show OLT1177 inhibits GSDMD cleavage to GSDMD-N in a dose-dependent manner. (B, C) Densitometric analysis of the GSDMD-N and IL-1β bands from . All concentrations of OLT1177 significantly reduce both GSDMD-N and IL-1β in a dose-dependent manner. D) Western blot shows NLRP3 and β-actin expression in BMDMs. Data are representative of three independent biological replicates (BMDMs derived from three independent mice). ****P< 0.0001.

Article Snippet: After 3.5 hours, the NLRP3 inflammasome formation was induced with 10 μM nigericin (InvivoGen, San Diego, CA) for 30 minutes.

Techniques: Western Blot, Expressing, Derivative Assay

Journal: bioRxiv

Article Title: OLT1177 (Dapansutrile) inhibits Gasdermin D-dependent IL-1β Release and Pyroptotic Cell Death in Bone Marrow-derived Macrophages

doi: 10.64898/2026.05.31.729061

Figure Lengend Snippet: LPS induces cell activation and expression of the NLRP3 gene. Nigericin is a polyether antibiotic and acts as a potassium ionophore, causing NLRP3 inflammasome activation. Caspase-1 cleaves pro-IL-1β and GSDMD to IL-1β and GSDM-N, respectively. Mature IL-1β then exits the cells through the GSDMD-N pores.

Article Snippet: After 3.5 hours, the NLRP3 inflammasome formation was induced with 10 μM nigericin (InvivoGen, San Diego, CA) for 30 minutes.

Techniques: Activation Assay, Expressing

Journal: JCI Insight

Article Title: Combined single-cell transcriptome and immune repertoire analysis reveals hepatic and renal immune injury by heat stroke

doi: 10.1172/jci.insight.189825

Figure Lengend Snippet: ( A ) Subclustering of myeloid cells identified 11 cell subclusters. ( B ) Dot plot illustrating the scaled expression of functional marker genes across each myeloid cell subcluster. ( C ) UMAP plot (left) and violin plot (right) displaying the expression of NLRP3 inflammasome feature scores in each myeloid cell subcluster. ( D ) UMAP plot (left) and violin plot (right) showing the expression of pyroptosis feature scores in each myeloid cell subcluster. ( E ) Bar plot illustrating the relative contribution of each of the 10 myeloid cell subclusters in different sample groups. P values were obtained using the 1-way Kruskal-Wallis test with post hoc Dunn’s test. ( F ) Dot plot showing the expression of inflammasome and pyroptosis features in myeloid cell subtypes derived from the HC, HE, and HS groups. ( G ) Ranking of significantly differentially expressed genes between HS-enriched and HC-enriched myeloid cells. ( H ) GSEA plot illustrating pathways enriched in HS-associated myeloid cell clusters compared with HC-associated clusters. NES, normalized enrichment score.

Article Snippet: The third and the fourth groups were intraperitoneally injected with NLRP3 inflammasome inhibitor (CY-09, Selleck) or TNF-α inhibitor QNZ (Selleck) for 7 days before the induction of the HS model, according to previous reports ( , ).

Techniques: Expressing, Functional Assay, Marker, Derivative Assay

Journal: JCI Insight

Article Title: Combined single-cell transcriptome and immune repertoire analysis reveals hepatic and renal immune injury by heat stroke

doi: 10.1172/jci.insight.189825

Figure Lengend Snippet: ( A ) Diagram of intervention procedures for HS induction in 4 groups. The third and fourth groups were intraperitoneally injected with the NLRP3 inhibitor CY-09 (3 mg/kg/day) or the TNF-α inhibitor QNZ (0.1 mg/kg/day) for 7 consecutive days before HS induction ( n = 10 per group). The other 2 groups received PBS as vehicle control. Mice were euthanized 2 hours after HS onset for sample collection. Serum samples were used for cytokine and biological function measurements. Hepatic and renal tissues were used for pathology staining and Western blotting. ( B ) Representative images of H&E-stained hepatic and renal tissues from the experimental groups. Original magnification, ×400. Scale bars: 50 μm. n = 3 repeats per group. ( C ) Clinical parameters measured include markers for hepatic function (ALT, AST), renal function (serum urea, creatinine), and rhabdomyolysis (creatine kinase); n = 9 repeats per group. Comparisons were conducted using the 1-way ANOVA followed by Tukey’s multiple-comparison test. ( D ) Key plasma cytokines (IL-6, IL-1β, IL-18, TNF-α) and chemokines (CCL2, CCL3, CCL4) quantified by ELISA; n = 9 repeats per group. Comparisons were conducted using 1-way ANOVA followed by Tukey’s multiple-comparison test. ( E and F ) Western blots and bar graphs displaying expression profiles of inflammasome- and pyroptosis-related proteins (NLRP3, TLR4, ASC, GSDMD, caspase 1, caspase 11) and inflammatory cytokines (IL-6, IL-1β, IL-18, TNF-α, MCP-1) in hepatic tissues, with β-actin as loading control; n = 3 repeats per group. Comparisons were conducted using 1-way ANOVA followed by Tukey’s multiple-comparison test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The third and the fourth groups were intraperitoneally injected with NLRP3 inflammasome inhibitor (CY-09, Selleck) or TNF-α inhibitor QNZ (Selleck) for 7 days before the induction of the HS model, according to previous reports ( , ).

Techniques: Injection, Control, Staining, Western Blot, Comparison, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: Inflammation and pyroptosis were increased in CP mice. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. The animals were sacrificed at 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A-C ) Representative photos and quantitative of F4/80 and CD11b staining; ( D ) Serum IL-6 level; ( E ) Serum TNF-α level; ( F & G ) Western blot analysis and quantitative of the expression of NLRP3 in the pancreas; ( H ) Serum IL-18 level; ( I ) Serum IL-1β level; ( J ) Correlation analysis of serum IL-18 and Masson positive aera; ( K ) Correlation analysis of serum IL-1β and Masson positive aera; ( L ) Correlation analysis of serum CIRP and F4/80 positive cells; ( M ) Correlation analysis of serum CIRP and CD11b positive cells; ( N ) Correlation analysis of serum CIRP and F4/80 positive cells; ( O ) Correlation analysis of serum CIRP and CD11b positive cells; ( R ) Correlation analysis of serum CIRP and IL-18; ( S ) Correlation analysis of serum CIRP and IL-1β; ( T ) Correlation analysis of serum CIRP and serum IL-18 in CP patients; ( U ) Correlation analysis of serum CIRP and serum IL-1β in CP patients; ( V ) Correlation analysis of serum CIRP and serum HMGB1 in CP patients. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Statistical significance was determined by t-test or one-way ANOVA with Tukey’s post-hoc test. Correlations were assessed using Spearman’s rank correlation coefficient. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; L-arg, L-arginine; Cer, Cerulein; IL-6, interleukin 6; TNF-α, tumor necrosis factor-α; NLRP3, Nod-like receptor family protein 3; IL-18, interleukin 18; IL-1β, HMGB1, high mobility group protein B1; interleukin 1β

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Staining, Western Blot, Expressing, RNA Binding Assay

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: CIRP deficiency alleviates inflammation and pyroptosis in CP model mice. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. 8 mg/kg C23 was administered 2 h after the last L-arginine or cerulein injection each time. To determine the role of TLR4 in CIRP’s effect in CP, TAK-242, a specific TLR4 receptor inhibitor, were administered through intraperitoneal injection at 1 h after the last injection of L-arginine or cerulein each time. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A & B ) Western blot analysis and quantitative of the expression of NLRP3, Cleaved Caspase-1 and Cleaved Gasdermin D in the pancreas; ( C-E ) Representative photos and quantitative of F4/80 and CD11b staining; ( F ) Serum IL-1β level; ( G ) Serum IL-18 level; ( H-J ) Western blot analysis and quantitative of the expression of NLRP3 and Cleaved Gasdermin D in the pancreas; ( K & L ) Representative photos and quantitative of F4/80 staining. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; KO, knockout; L-arg, L-arginine; NLRP3, Nod-like receptor family protein 3; TLR4, Toll-like receptor 4; IL-18, interleukin 18; IL-1β, interleukin 1β; Cer, Cerulein

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Western Blot, Expressing, Staining, RNA Binding Assay, Knock-Out

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: NLRP3 inhibitor alleviates inflammation and pancreatic fibrosis in Experimental CP. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. At 2 h after the last injection of L-arginine or cerulein each time, normal saline (vehicle) or 5, 20, 100 mg/kg NLRP3 inhibitor was administered by intraperitoneal injection. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A ) Western blot analysis of the expression of CIRP, NLRP3, Cleaved Caspase-1 and Cleaved Gasdermin D in the pancreas; ( B ) Serum IL-1β level; ( C ) Serum IL-6 level; ( D-G ) Representative photos and quantitative of H&E, Sirius red and F4/80 staining; ( H ) Serum HMGB1 level. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; NLRP3, Nod-like receptor family protein 3; IL-6, interleukin 6; IL-1β, interleukin 1β; HMGB1, high mobility group protein B1; Cer, Cerulein; L-arg, L-arginine

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Saline, Western Blot, Expressing, Staining, RNA Binding Assay

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: Gasdermin D inhibitor alleviates inflammation and pancreatic fibrosis in Experimental CP. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. At 2 h after the last injection of L-arginine or cerulein each time, normal saline (vehicle) or 5, 20, 50 mg/kg Disulfiram, a potent gasdermin D inhibitor, was administered by intraperitoneal injection. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A-D ) Representative photos and quantitative of H&E, Sirius red and F4/80 staining; ( E ) Western blot analysis of the expression of CIRP, NLRP3 and Cleaved Gasdermin D in the pancreas; ( F ) Serum IL-18 level. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; NLRP3, Nod-like receptor family protein 3; IL-18, interleukin 18

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Saline, Staining, Western Blot, Expressing, RNA Binding Assay