Journal: Journal of Agricultural and Food Chemistry
Article Title: Natural Dietary Flavonoid Apigenin Mitigates Ulcerative Colitis via Modulating the AMPK/NF-κB/NLRP3 Signaling Axis
doi: 10.1021/acs.jafc.5c15405
Figure Lengend Snippet: Api inhibits both in vivo and in vitro activation of NLRP3 inflammasome. (A) Relative mRNA expression of Nlrp3 in colonic tissues from Control, DSS, and DSS + Api groups (qPCR; n = 5). (B) Western-blot analysis of inflammasome-related proteins in colonic tissue: cleaved and pro-IL-1β, cleaved and pro-caspase-1, NLRP3, ASC, GSDMD; β-actin was used as a loading control. (C) ELISA quantification of IL-1β in BMDM supernatants after LPS + nigericin stimulation with or without Api ( n = 3). (D) qPCR analysis of IL-1β , Nlrp3 , and GSDMD transcripts in BMDMs under the same conditions as in (C) ( n = 3). (E) SPR sensorgram confirming the direct interaction between Api and recombinant NLRP3. (F) Molecular-docking model showing the predicted binding of Api to the NACHT domain of NLRP3 (PDB: 7ALV); calculated binding energy, −8.6 kcal/mol. ### p < 0.001 versus Control; * p < 0.05, ** p < 0.01, *** p < 0.001 versus DSS.
Article Snippet: Commercially available recombinant human NLRP3 protein (CSB-EP822275HU7) and AMPK α1/β2/γ1 heterotrimer protein (HY- P76143 , MCE) were immobilized (∼3,000 RU) on a CM5 Chip (GE Healthcare, Piscataway, NJ, USA) according to a standard amine coupling procedure.
Techniques: In Vivo, In Vitro, Activation Assay, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay