Fig 1: The inflammatory factor levels were suppressed by Celastrol. (a) The level of TNF-𝑎 was measured. (b) The level of IL-31 was measured. (c) The level of IL-6 was measured. (d) The level of IL-1β was measured. ∗ suggests p < 0.05.
Fig 2: Effects of DLE and MC on IL-6, IL-31 and IL-33 expression in LPS-stimulated microglia. Microglia were pretreated with DLE or MC at the indicated concentrations and stimulated with LPS for 24 or 3 h. (A) IL-6, IL-31 and IL-33 mRNA expression levels were determined by reverse transcription PCR. (B) IL-6, IL-31 and IL-33 protein expression levels were determined by western blot analysis. Each bar represents the mean ± SD (n=3). #P<0.05 vs. untreated cells; *P<0.05 vs. LPS alone treated cells. DLE, Diospyros lotus leaf extract; MC, myricitrin; LPS, lipopolysaccharide.
Fig 3: Effect of IL-6 and IL-31 gene silencing on LCN2 and STAT3 activation of astrocytes by LSMCM. Astrocytes were cultured in stimulated microglia conditioned media for 24 h. Prior to receiving LPS-stimulation, microglial cells were silenced for IL-6 or IL-31 or IL-6 plus IL-31 genes. (A) Relative expression levels of LCN2 and STAT3 were evaluated by western blot analysis. Transfection efficiency was confirmed by reverse transcription PCR. (B) IL-6 mRNA and (C) IL-31 mRNA expression levels. Each bar represents the mean ± SD (n=3). #P<0.05 vs. untreated cells; *P<0.05 vs. LSMCM alone treated cells; ns, no significant difference compared to LSMCM alone treated cells; DLE, Diospyros lotus leaf extract; MC, myricitrin; LSMCM, LPS-stimulated microglia culture medium; LPS, lipopolysaccharide; LCN2, lipocalin-2; si, short interfering.
Fig 4: Celastrol has a therapeutic effect on AD‐like symptoms and degranulation of mast cells in Balb/c mice. (A) Toluidine blue staining was used to observe the infiltration and degranulation of mast cells in the dorsal skin and ear skin of mice. The mast cell infiltration, degranulation number and degranulation rate of mast cells were shown on the lower panel. (B) Detection of Th2 cytokines IL‐4, IL‐13 and TNF‐α in mouse lymph nodes by flow cytometry. (C) Detection of Th2 cytokines IL‐4, IL‐13 and TNF‐α in mouse spleen cells by flow cytometry. (D) ELISA detection of chemokines CCL11, CCL17 and cytokine IFN‐γ in mouse skin homogenate, as well as the expression of IL‐1β, TSLP, IL‐17, IL‐33, IL‐5, IL‐31, IL‐23 and IgE. *p < 0.05 and **p < 0.01, compared to the Control group; and #p < 0.05 and ##p < 0.01, compared with the Model group.
Supplier Page from Abcam for Mouse IL-31 ELISA Kit