Journal: Frontiers in Pharmacology

Article Title: Piperlongumine Is an NLRP3 Inhibitor With Anti-inflammatory Activity

doi: 10.3389/fphar.2021.818326

Figure Lengend Snippet: PL disrupts NLRP3 inflammasome activation in vivo . (A–E) Effects of PL in endotoxemia. Wild-type mice with the C57BL/6 background were pretreated with intraperitoneal injection of PL (50 mg/kg or 100 mg/kg) or N.S 30 min before intraperitoneal injection of LPS (20 mg/kg) for 8 h. ELISA analysis of IL-1β (A) , IL-6 (B) , and TNF-α (C) was performed in mice blood serum. (D) Lung W/D ratio of endotoxemic mice treated with PL (50 mg/kg or 100 mg/kg). (E) Representative images of HE staining in lungs. (F, G) Effects of PL in MSU-induced peritonitis. Wild-type mice with the C57BL/6 background were intraperitoneally injected of PL (100 mg/kg) or N.S 30 min before intraperitoneal injection of MSU (1 mg) for 6 h. ELISA of IL-1β (F) and neutrophils (flow cytometry) (G) in the peritoneal cavity fluid were quantified. Values presented are mean ± SD. Two-way ANOVA and the Bonferroni test were used for the statistical analysis; *** p < 0.001; **** p < 0.0001. Data were collected from three independent experiments.

Article Snippet: Standard LPS (E. coli 0111:B4, Cat No. tlrl-eblps), ultrapure LPS (E. coli 0111:B4, Cat No. tlrl-3pelps), nigericin (Cat No. tlrl-nig), ATP (Cat No. tlrl-atpl), and MSU (Cat No. tlrl-msu) were purchased from InvivoGen (San Diego, CA, United States); the cell lysis buffer (CLB) (Cat No. 9803) was bought from Cell Signaling Technology (Danvers, MA, United States); the mouse immunoglobin IgG protein (Cat No. ab198772) was purchased from Abcam (Cambridge, CB2 0AX, United Kingdom); Protein A/G PLUS-Agarose (Cat No. sc-2003) was obtained from Santa Cruz (Santa Cruz, CA, United States); mouse IL-1β (Cat No. 88–7013), tumor necrosis factor-α (TNF-α) (Cat No. 88-7324), interleukin-6 (IL-6) (Cat No. 88-701364), and a human IL-1β (Cat No. BMS22) ELISA kit was bought from Thermo Fisher (Waltham, MA United States); and the CellTiter-Glo ® Luminescent Cell Viability Assay (Cat No. G7572) was from Promega.

Techniques: Activation Assay, In Vivo, Injection, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry