Betaine Modulates NF-κB and NLRP3 Pathways

Betaine (molecular formula: C₅H₁₁NO₂, CAS No.: 107-43-7, purity >98%) was procured from Shanghai Yuanye Bio-Technology Co., Ltd, China. AβO was purchased from China, Peptides Co. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was obtained from Sigma-Aldrich, USA. The NF-κB activator phorbol 12-myristate 13-acetate [19 (link)] (PMA, a protein kinase C activator that can induce NF-κB activation in BV2 cells; this concentration was confirmed in previous studies; molecular formula: C36H56O8, CAS No.: 16561-29-8, purity >98%) was procured from MedChemExpress, China. A protein quantification (BCA assay) kit was provided by Beyotime Institute of Biotechnology, China. Antibodies against NF-κB p65, NLRP3, ASC, and Caspase-1 were provided by Cell Signaling Technology (USA). Enzyme-linked immunosorbent assay (ELISA) kits were procured from CUSABIO Technology, China (IL-1β ELISA kit: detection range: 31.25 pg/mL-2000 pg/mL; IL-18 ELISA kit: detection range: 1.56 pg/mL-100 pg/mL; TNF-α ELISA kit: detection range: 15.6 pg/mL-1000 pg/mL. Kit intra-assay precision: CV% <8%, interassay precision: CV% <10%).