Fig 1: M1 and M2 inflammation are evident in ovarian stroma with obesity and aging, with M2 macrophage phenotype reduced in BGP-15–treated mice.(A) Gene expression of inflammatory cytokines (Tnfa and Il6) in ovarian stromal cells of control (C), obese, and old mice treated with BGP-15 (B) or vehicle (v). Expression was normalized to L19 and was presented relative to controls. Values represent means ± SEM of n = 3 replicate pools of stromal cells from multiple mice. (B) TNFα and IL-6 (normalized to total protein) in ovarian stromal cells. Values represent means ± SEM of n = 3 to 4 replicate samples of material pooled from multiple mice. (C) Gene expression of Ifng and anti-inflammatory cytokines (Tgfb1, Il4, and Il13) in ovarian stromal cells from (A). (D) Localization of macrophages in the ovarian stroma using anti-F4/80 antibody (red; macrophages) and anti–4-HNE antibody (green; lipid peroxidation). Scale bars, 100 μm. (E) Numbers of F4/80+ (left) or 4-HNE+ (right) cells in the ovarian stroma. (F) Localization of iNOS (marker of M1 macrophage; green) and CD163 (marker of M2 macrophage; red) in ovarian stroma by immunohistochemistry. Scale bars, 100 μm. (G) Number of iNOS+ or CD163+ cells within the area of ovarian stroma. *P < 0.05 and **P < 0.01 by one-way ANOVA compared with controls or by two-tailed t test compared to vehicle-treated mice as indicated.
Fig 2: Effect of PDJ on the LPS-induced pro-inflammatory cytokine expression levels in RAW264.7 cells. RAW264.7 cells were pretreated with LPS (1 µg/ml) at 37°C for 1 h and then subsequently treated with PDJ (0, 2.5 or 5 µg/ml) for at 37°C 24 h. mRNA expression levels of the pro-inflammatory cytokines (A) IL-1β, (B) IL-6 and (C) TNFα were measured by reverse transcription-quantitative PCR. Data are presented as the mean ± SEM of three independent experiments. ###P<0.001 vs. untreated group; *P<0.05 vs. LPS-only treated group; **P<0.01 vs. LPS-only treated group; ***P<0.001 vs. LPS-only treated group. PDJ, polyphenolic extract from the Dowijigi flower; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor.
Fig 3: Schematic representation of the NF-κB-mediated inhibition of inflammatory responses by the polyphenolic extract of Dowijigi flower. LPS, lipopolysaccharide; TLR, Toll-like receptor; IκB, inhibitor of κB; IκBα, inhibitor of κBα; COX2, cyclooxygenase-2; iNOS, inducible nitric oxide synthase; IL, interleukin; TNF, tumor necrosis factor; NO, nitric oxide; PGE2, prostaglandin E2; p, phosphorylated.
Supplier Page from Enzo Life Sciences, Inc. for TNF-α (mouse), ELISA kit