Fig 1: Effect of PALO and/or MLA on hippocampal contents of (a) cleaved caspase-1, (b) IL-1β, (c) IL-18, and (d) caspase-11, as well as (e) ACh and (f) 5-HT in HFFD/LPS-induced AD model. Values are shown in scatter plots and expressed as mean ± SD (n = 5–6/group). Panel (a) depicts representative cropped blot of cleaved caspase-1 and its densitometric analysis (n = 5/group). Cleaved caspase-1, caspase-11 and 5-HT were analyzed using one-way ANOVA followed by Tukey’s post hoc test (n = 6/group), while Welch’s ANOVA was performed followed by Dunnett’s T3 Multiple Comparison test for IL-1β, IL-18 and ACh to account for heterogeneity (n = 6/group); p < 0.05. The symbols (□) and (○) indicate additive and synergistic interactions, respectively, using coefficient drug index (CDI). PALO (0.1 mg·kg−1; i.p) and/or MLA (5.6 mg·kg−1; i.p) were administered 3 h after LPS injection and for 8 days. 5-HT: serotonin; ASC/TMS1: apoptosis-associated speck-like protein/the target of methylation-induced silencing-1; IL-18: interleukin 18.
Fig 2: Inflammasome-associated cytokines.IL‐1β and IL‐18 protein levels in response to CLP. Plasma (A and D), kidney (B and E), and lung (C and F) IL‐1β and IL‐18 were measured via ELISA in Sham, CLP, and CLP rats treated with MCC950 (n = 12/group). *P < 0.05 versus Sham *p<0.05 vs Sham, #p<0.05 vs CLP.
Fig 3: (A) Western blot assay for detecting the expression of caspase-1, IL-1β, and IL-18. (B) Quantification of the expression levels of caspase-1, IL-1β, and IL-18. (C) ELISA assay for quantifying the expression levels of caspase-1, IL-1β, and IL-18 in the sham, Aβ1-42, Aβ1-42+LPS, and Aβ1-42+LPS+TTP488 groups. (D) Western blot assay for detecting the expression of caspase-1, IL-1β, and IL-18. (E) Quantification of the expression levels of caspase-1, IL-1β, and IL-18. (F) ELISA assay for quantifying the expression levels of caspase-1, IL-1β, and IL-18 in the sham, Aβ1-42, Aβ1-42+control oeNLRP1, Aβ1-42+oeNLRP1, and Aβ1-42+oeNLRP1+TTP488 groups. Protein levels were normalized to those of β-actin (Aβ1-42+LPS or Aβ1-42+oeNLRP1 vs. Aβ1-42 group, **p<0.05; Aβ1-42+LPS+TTP488 or Aβ1-42+oeNLRP1+TTP488 vs. Aβ1-42+LPS or Aβ1-42+oeNLRP1 group, ## p<0.05, n=6 per group).
Fig 4: Inflammasome-associated cytokines. Plasma (A,D), kidney (B,E), and lung (C,F) IL-1β and IL-18 were measured in Sham, CLP, and CLP rats treated with Clopidogrel (n = 12/group) via ELISA. * p < 0.05 vs. Sham, # p < 0.05 vs. CLP, n.d. = not detected.
Fig 5: Methylene blue inhibits the production of mature IL-1β and IL-18 by microglia. (A) Gating strategy for distinguishing and sorting microglia form normal rat spinal cords. CD45lowCD11b+ cells were microglia. N = 3 per group. (B) Apoptosis of microglia cultured for 48 h. V: vehicle phosphate bufferedsaline (PBS); 10: 10 nM methylene blue; 1000: 1000 nM methylene blue. Numbers in the quadrats are percentage of each gated population. N = 3 per group. (C,D) Concentrations of IL-1β and IL-18 in the supernatants. Un: no stimulation. lipopolysaccharide (LPS)+adenosine triphosphate (ATP): stimulation with LPS followed by ATP treatment. V: vehicle; 10~1000: methylene blue concentrations (nM). *p < 0.05; **p < 0.01 in comparison with stimulated cells of the vehicle group. (E–G) mRNA levels of IL-1β, IL-18 and TNF-α in cultured microglia. V: vehicle; M: 500 nM methylene blue. ***p < 0.001 in comparison with unstimulated cells of the vehicle group. N = 6–8 per group.
Supplier Page from Abcam for Rat IL-18 ELISA Kit