gsdmd inhibition  (MedChemExpress)

Journal: Cell Communication and Signaling : CCS

Article Title: NLRP3-mediated epithelial pyroptosis involved in airway hyperresponsiveness in combined eosinophilic and neutrophilic asthma

doi: 10.1186/s12964-026-02706-5

Figure Lengend Snippet: Evidence of cell pyroptosis existed in EOS + NEU + inflammation asthma. a Protein levels of NLRP3, cleaved caspase-1 and GSDMD-NT were upregulated in HBE after 24 h stimulation with IL-13 and IL-17 A. b , c IL-1β and LDH levels in cell culture supernatants were measured ( n = 6 per group). d Scanning electron microscopy revealed pore formation on the surface of HBEs stimulated with IL-13 and IL-17 A, scale bar, 5 μm. e , f Expression of GSDMD-NT and cleaved caspase-1 was elevated in lung tissues of mice exposed to OVA and ozone, accompanied by the dysfunction of cell junctions. Scale bar, 500 nm. g - i mRNA expression levels of caspase-1, GSDMD and IL-1β were increased. Data were expressed as mean ± SD. Abbreviations: Ctrl, control; EOS, eosinophil; GSDMD, Gasdermin D; GSDMD-NT, Gasdermin D-N-terminal; HBE, human bronchial epithelial cells; IL, interleukin; LDH, lactate dehydrogenase; NEU, neutrophil; NLRP3, NOD-like receptor protein 3; OVA, ovalbumin; SD, standard deviation

Article Snippet: For GSDMD inhibition, disulfiram (50 mg/kg in corn oil; HY-B0240, MedChemExpress, Shanghai, China) or vehicle was administered via intraperitoneal injection before each OVA challenge, following established protocols [ , ].

Techniques: Cell Culture, Electron Microscopy, Expressing, Control, Standard Deviation

Journal: Cell Communication and Signaling : CCS

Article Title: NLRP3-mediated epithelial pyroptosis involved in airway hyperresponsiveness in combined eosinophilic and neutrophilic asthma

doi: 10.1186/s12964-026-02706-5

Figure Lengend Snippet: NLRP3 inhibition alleviates AHR and suppresses GSDMD activation. a , b AHR was reduced in NLRP3 −/− mice after OVA and ozone exposure. c - g HE and PAS staining revealed attenuated airway inflammation and mucus hypersecretion in NLRP3 −/− mice. h Immunofluorescence showed decreased GSDMD-NT expression in lung tissues, scale bars 100 µm. i - l Inflammatory cytokine levels and caspase-1 in BALF or lung tissue were measured using ELISA. m Treatment with MCC950 reduced the expression of NLRP3, cleaved caspase-1 and GSDMD-NT in HBE. n Immunofluorescence demonstrated changes in NLRP3 and GSDMD-NT expression in HBEs treated with MCC950, scale bars 200 µm. Data were expressed as mean ± SD. Abbreviations : AHR Airway hyperresponsiveness, BALF Bronchoalveolar lavage fluid, Ctrl , Control, DAPI 4’,6-diamidino-2-phenylindole, ELISA Enzyme-linked immunosorbent assay, GSDMD Gasdermin D, GSDMD-NT Gasdermin D-N-terminal, HBE Human bronchial epithelial cells, HE Haematoxylin and eosin, NLRP3 NOD-like receptor protein 3, OVA Ovalbumin, PAS Periodic acid-Schiff

Article Snippet: For GSDMD inhibition, disulfiram (50 mg/kg in corn oil; HY-B0240, MedChemExpress, Shanghai, China) or vehicle was administered via intraperitoneal injection before each OVA challenge, following established protocols [ , ].

Techniques: Inhibition, Activation Assay, Staining, Immunofluorescence, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Cell Communication and Signaling : CCS

Article Title: NLRP3-mediated epithelial pyroptosis involved in airway hyperresponsiveness in combined eosinophilic and neutrophilic asthma

doi: 10.1186/s12964-026-02706-5

Figure Lengend Snippet: Inhibition of GSDMD alleviates mixed airway inflammation and AHR. a - d Lung function variables improved following disulfiram treatment ( n = 6 per group). e , f Disulfiram significantly reduced AHR in EOS + NEU + asthmatic mice. g - i HE and PAS staining analysis demonstrated reduced airway inflammation and mucus secretion after GSDMD inhibition. j - l Levels of inflammatory cytokines in BALF were decreased following disulfiram treatment; ( g , h ) HBEs were transfected with GSDMD small interfering RNA (siRNA) for 24 h and were then treated with IL-13/IL17A for another 24 h. The expression of IL-1β was studied using ELISA analysis. Data were expressed as mean ± SD. Abbreviations: AHR Airway hyperresponsiveness, BALF Bronchoalveolar lavage fluid, ELISA Enzyme-linked immunosorbent assay, GAPDH Glyceraldehyde-3-phosphate dehydrogenase, GSDMD Gasdermin D, HBE Human bronchial epithelial cells, HE Haematoxylin and eosin, IL Interleukin, OVA Ovalbumin

Article Snippet: For GSDMD inhibition, disulfiram (50 mg/kg in corn oil; HY-B0240, MedChemExpress, Shanghai, China) or vehicle was administered via intraperitoneal injection before each OVA challenge, following established protocols [ , ].

Techniques: Inhibition, Staining, Transfection, Small Interfering RNA, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 deficiency protects against acetaminophen‑induced liver injury by inhibiting hepatocyte pyroptosis

doi: 10.3892/mmr.2024.13185

Figure Lengend Snippet: APAP overdose induces hepatic NLRP3 activation in mice. C57BL/6 mice were intraperitoneally injected with 300 mg/kg APAP at the indicated time points (n=5). The histopathological dynamic changes of liver tissues shown by (A) H&E staining in liver tissues and (B) statistical quantification of hepatic necrosis area. Serum levels of (C) ALT and (D) AST. (E) IHC staining of NLRP3, caspase-1, IL-1β and GSDMD in liver tissues. (F) Western blotting of NLRP3, pro-cas-1, cl-cas-1, pro-IL-1β, cl-IL-1β, GSDMD and GSDMD-N in liver tissues. Serum levels of (G) IL-1β and (H) IL-18. (I) Mouse liver parenchymal cells were isolated after 24 or 48 h post-treatment with APAP for western blotting of NLRP3, pro-cas-1, cl-cas-1, pro-IL-1β, cl-IL-1β, GSDMD and GSDMD-N. Data are presented as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001. pro-cas-1, pro-caspase-1; cl-cas-1, cl-caspase-1; GSDMD, gasdermin D; GSDMD-N, cleaved GSDMD N-terminal fragment; cl-IL-1β, cleaved IL-1β; APAP, acetaminophen; NLRP3, NLR family pyrin domain containing 3; ALT, alanine aminotransferase; AST, aspartate transaminase; con, control; IHC, immunohistochemistry.

Article Snippet: To evaluate the therapeutic effects of NLRP3 and GSDMD inhibition, NLRP3 inhibitor MCC950 (TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 2 h after APAP and GSDMD inhibitor disulfiram (DSF; TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 4 and 24 h before APAP (n=5 per group), with PBS or vehicle (sesame oil) as control group respectively.

Techniques: Activation Assay, Injection, Staining, Immunohistochemistry, Western Blot, Isolation, Control

Journal: Molecular Medicine Reports

Article Title: NLRP3 deficiency protects against acetaminophen‑induced liver injury by inhibiting hepatocyte pyroptosis

doi: 10.3892/mmr.2024.13185

Figure Lengend Snippet: APAP induces NLRP3 activation and pyroptosis in hepatocytes in vitro (n=3). AML12 cells and mouse primary hepatocytes were stimulated with different concentrations of APAP for 24 h or 12 h. (A) Viability of both cell types. RT-qPCR analysis of (B) Nlrp3 , (C) caspase-1 and (D) Il-1β in these two cell types. (E) Western blotting and (F) quantification of NLRP3, pro-cas-1, cl-cas-1, pro-IL-1β, cl-IL-1β, GSDMD and GSDMD-N in both cell types. Data are presented as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001. pro-cas-1, pro-caspase-1; cl-cas-1, cl-caspase-1; GSDMD, gasdermin D; GSDMD-N, cleaved GSDMD N-terminal fragment; APAP, acetaminophen; NLRP3, NLR family pyrin domain containing 3; ALT, alanine aminotransferase; AST, aspartate transaminase; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: To evaluate the therapeutic effects of NLRP3 and GSDMD inhibition, NLRP3 inhibitor MCC950 (TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 2 h after APAP and GSDMD inhibitor disulfiram (DSF; TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 4 and 24 h before APAP (n=5 per group), with PBS or vehicle (sesame oil) as control group respectively.

Techniques: Activation Assay, In Vitro, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: NLRP3 deficiency protects against acetaminophen‑induced liver injury by inhibiting hepatocyte pyroptosis

doi: 10.3892/mmr.2024.13185

Figure Lengend Snippet: NLRP3 inhibitor MCC950 alleviates APAP-induced liver injury and hepatocyte pyroptosis. C57BL/6 mice were treated with MCC950 or PBS following APAP treatment (n=5). The therapeutic effect of MCC950 in APAP induced liver injury, with PBS treated mice as control. (A) Representative images of H&E staining and the statistical quantification of hepatic necrosis in each group. (B) Representative images of TUNEL staining and the statistical quantification of TUNEL-positive cells/field in two groups. Serum (C) ALT and (D) AST levels for two groups. (E) Viability of AML12 cells after co-treatment with APAP and different concentrations of MCC950 for 24 h. (F) Western blotting of cl-cas-1, cl-IL-1β, GSDMD-N and the statistical quantification of each protein level in AML12 cells after co-treatment with 10 mM APAP and 0, 1, 5, 10 µM MCC950 for 24 h. Data are presented as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001. APAP, acetaminophen; NLRP3, NLR family pyrin domain containing 3; cl-cas-1, cl-caspase-1; GSDMD-N, cleaved gasdermin D N-terminal fragment; cl-IL-1β, cleaved IL-1β.

Article Snippet: To evaluate the therapeutic effects of NLRP3 and GSDMD inhibition, NLRP3 inhibitor MCC950 (TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 2 h after APAP and GSDMD inhibitor disulfiram (DSF; TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 4 and 24 h before APAP (n=5 per group), with PBS or vehicle (sesame oil) as control group respectively.

Techniques: Control, Staining, TUNEL Assay, Western Blot

Journal: Molecular Medicine Reports

Article Title: NLRP3 deficiency protects against acetaminophen‑induced liver injury by inhibiting hepatocyte pyroptosis

doi: 10.3892/mmr.2024.13185

Figure Lengend Snippet: GSDMD inhibitor DSF mitigates APAP-induced liver injury and hepatocyte pyroptosis. C57BL/6 mice were treated with DSF or vehicle prior to APAP treatment (n=5). (A) Representative images of H&E staining and the statistical quantification of hepatic necrosis. (B) IHC staining of hepatic Caspase-3 and the statistical quantification of the positive area. Serum (C) ALT and (D) AST levels for two groups. (E) Western blotting of GSDMD-N, Pro-Cas-3/Cl-Cas-3 and the statistical quantification of each protein level in AML12 cells after co-culture with 10 mM APAP and 0, 1, 2.5, 5 µM DSF for 24 h. Data were presented as the mean ± SD. **P<0.01, ***P<0.001. APAP, acetaminophen; cl-cas-1, cl-caspase-1; pro-cas-1, pro-caspase-1; GSDMD, gasdermin D; GSDMD-N, cleaved GSDMD N-terminal fragment, DSF. disulfiram; IHC, immunohistochemical staining; ALT, alanine aminotransferase; AST, aspartate transaminase.

Article Snippet: To evaluate the therapeutic effects of NLRP3 and GSDMD inhibition, NLRP3 inhibitor MCC950 (TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 2 h after APAP and GSDMD inhibitor disulfiram (DSF; TargetMol Chemicals Inc.) was administered IP into mice at 50 mg/kg 4 and 24 h before APAP (n=5 per group), with PBS or vehicle (sesame oil) as control group respectively.

Techniques: Staining, Immunohistochemistry, Western Blot, Co-Culture Assay, Immunohistochemical staining