Expression profiles of NLRP3, ASC, and inflammatory cytokines in MDMs and sera from patients with type 2 diabetes (T2D) and
healthy controls (HCs). A, C–E: MDMs were obtained from 47 patients with T2D and 57 HCs. The MDMs were incubated with media containing autologous sera and
stimulated with ultrapure LPS (A, C, and E; 10 ng/mL) and Pam3CSK4 (Pam, E; 10 ng/mL) for 4 h. B: MDMs were cultured from T2D patients (n = 10) and HCs (n = 10). The protein levels of NLRP3 and ASC were measured by Western blot analysis. The intensity of each band for each protein
was quantified and normalized relative to the housekeeping gene β-actin (B, right). D: Sera collected from the peripheral blood of T2D patients (n = 47) and HCs (n = 57). The mRNA expression of Il1β, Nlrp3, and Asc (A) and Il6, Il8, Tnfα, and Ccl2 (E) was analyzed by quantitative real-time RT-PCR. IL-1β and IL-18 production in culture supernatants (C) and sera (D) were measured by ELISA. Serum cytokine levels were determined in samples pretreated with protease inhibitors (4% volume/volume).
A and E: Results are expressed as the mean concentration of triplicate samples. Data are representative of two independent experiments.
B–D: Data are expressed as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. HCs. U, untreated.