Fig 1: HET0016 as a lipid peroxidation inhibition protects against sepsis.A Levels of plasma MDA from healthy control or patients. B Spearman correlation analysis for plasma MDA and D-lactate or APACHE II score in all patients (n = 28). C Kaplan–Meier analysis according to a cutoff value calculated from ROC analysis. D Representative histopathological section images, immunohistochemical or fluorescent images in the intestinal tissues of patients with sepsis. The scale bar represents 50 µm. The quantified fluorescence intensity of 4-HNE was shown in the lower left corner. E Survival analysis of the indicated mice in CLP-induced sepsis with or without treatment of 10 mg/kg HET0016, 20 mg/kg JSH-23, or 10 mg/kg Carnosol at 2 h before CLP and 12, 24, 48, and 72 h after CLP (n = 15 per group). F Immunoblot analysis of p-TBK1, TBK1, p-P65, and P65 in PBMCs from patients with sepsis or healthy control, supplemented with or without HET0016 (5 µM) for 24 h (n = 4 per group). G qPCR analysis of Il-6, Il-1b, and Tnf mRNA in PBMCs from patients with sepsis or healthy control, supplemented with or without HET0016 (5 µM) for 24 h (n = 6 per group). H Production of MDA in PBMCs from patients with sepsis supplemented with or without HET0016 (5 µM) for 24 h (n = 6 per group). Data are shown as mean ± SD, and analysis by one-way ANOVA test. Data in (G, H) are analyzed by paired Student’s t-test. PBMCs peripheral blood mononuclear cells, MODS multiple organ dysfunction syndromes, MDA malondialdehyde, FTH ferritin heavy chain, GPX4 glutathione peroxidase 4, 4-HNE 4-hydroxynonenal. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001.
Fig 2: Dose- and time-dependent TGF-ß1-induced production of TNF-a and IL-6 in mouse podocyte cells. (A-D) Enzyme-linked immunosorbent assays were performed to determine the relative levels of soluble TNF-a (A and B) and IL-6 (C and D) in culture supernatants of podocytes treated with 12 ng/ml TGF-ß1 for the indicated periods (A and C) or treated with TGF-ß1 at the indicated doses for 72 h (B and D). All data are presented as mean ± SD of three independent experiments. **P<0.01. TGF, transforming growth factor; TNF-a, tumor necrosis factor-a; IL-6, interleukin-6.
Fig 3: Curcumin reprogrammed M2 macrophages into an M1-like population in IL-4-induced model. Raw264.7 cells were pretreated with IL-4 (20 ng/mL) for 24 h and then stimulated by curcumin (20 µM). (A,B). The relative mRNA expressions of M1 macrophage markers TNF-a, IL-6, IL-12, CD86, and iNOS and M2 macrophage markers IL-10, CD206, TGF-ß, and Arg-1 were measured by qRT-PCR assay. (C–E). The protein levels of Arg-1 and iNOS were evaluated through Western blot. (F). The secretion of IL-6, TNF-a, and Il-10 was measured by ELISA. (G–J). The percentage of the CD86 (G,H) and CD206 (I,J) positive populations in Raw264.7 cells. Data are presented as the mean ± SD (n = 3). p values were determined by Student’s t-tests or analysis of variance (ANOVA). * p < 0.05; ** p < 0.01; *** p < 0.001.
Fig 4: HDACi inhibited the cytokine expression in 3T3-L1 adipocytes.3T3-L1 cells were treated with TNF-a (50 ng/mL) combined with or without NaB(1 mM) or TSA(0.5 µM) for 8 hr. Total RNA was isolated for realtime RT-PCR to detect the gene expression of IL-1ß and IL-6 (A). Supernatants of cell culture were harvested for ELISA assay to detect IL-1 and IL-6 in the culture medium (B). N: Normal control. *P < 0.05 by t-test (TNF vs NaB or TSA).
Fig 5: SNHG1 interacts with HMGB1 in RAW264.7 cells. (A) The results of the silver staining assay showed the SNHG1 binding protein. (B) GO function analysis of differentially expressed proteins that bind SNHG1. (C) KEGG pathway analysis of differentially expressed proteins that bind SNHG1. (D) Two HMGB1 peptides were detected in the mass spectrometry results. (E, F) The expression of HMGB1 in RAW264.7 cells treated with LPS, as determined by Western blot. (G) Anti-HMGB1 RIP assay was executed in RAW264.7 cells, followed by qRT-PCR. (H) Expression of proinflammatory cytokines (TNF-a, iNOS, IL-18, MCP-1, IL-6 and IL-1ß) in RAW264.7 cells transfected with lentivirus sh-HMGB1 treated with LPS, as determined by qRT-PCR. (I, J) Expression of proinflammatory cytokines (TNF-a, IL-6) in RAW264.7 cells transfected with lentivirus sh-HMGB1 treated with LPS, as determined by ELISA. Data are indicated as the mean ± SD, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Supplier Page from CUSABIO Technology LLC for Mouse Interleukin 6,IL-6 ELISA Kit