Figure 1 YDZG inhibits NLRP3 inflammasome activation. (A)
Chemical structure of Yadanzigan (YDZG). (B) THP-1 cells were pretreated
with 40 μM YDZG for 1 h before being stimulated with LPS (1
μg·mL-1) for another 6 h, and treated with ATP (5 mM)
for 30 min before harvesting. Proteins were isolated and
immunoprecipitated with an antibody against ASC (n=5). (C) THP-1 cells
were pretreated with the indicated concentrations of YDZG for 1 h before
being stimulated with LPS for another 2 h, and were then treated with
ATP (5 mM) for 30 min. Caspase-1 p20 was determined using the
immunofluorescence assay described in the Methods section (n = 3). The
primary antibody anti-caspase-1 p20 (1:200) and secondary antibody goat
anti-Rabbit Alexa Fluor 488 (1:200) were employed (scale bar = 10 μm).
(D) Immunoblot analysis of supernatants (SN) or cell lysates (CL) of
THP-1 cells, primed with LPS, followed by stimulation with ATP for 30
min (n=5). THP-1 cells were pretreated with the indicated concentrations
of YDZG for 1 h before being stimulated with LPS for another 6 h and
then treated with ATP (5 mM) for 30 min. The expression of IL-1β, was
determined using ELISA (E) and qRT-PCR (F) (n=6). Data are reported as
mean ± SEM; *P < 0.05 compared with the LPS alone
group.