Journal: Journal of Inflammation Research

Article Title: NHWD-870, a Potent BET Inhibitor, Ameliorated Endotoxemia-Induced Hepatic Inflammation via Suppression of BRD4-STAT1 Signaling and Macrophage M1 Polarization

doi: 10.2147/JIR.S598139

Figure Lengend Snippet: Endotoxemia liver inflammation model was successfully established. ( A) Schematic diagram of LPS modeling (7.5mg/kg). (B) HE staining of liver tissue (magnification 100×). (C and D) Levels of serum ALT( C ) and AST( D ). (E–G) ELISA was used to detect the levels of serum IL6 ( E ), TNFα ( F ), and MCP1 ( G ). (H–J) RT-qPCR was used to detect the mRNA expression of IL6 ( H ), TNFα( I ), and MCP1 ( J ) in the liver tissues of mice. Mice were divided into 3 groups: Saline (n=5), LPS 12h (n=5), LPS 24h (n=5). The data are presented as mean ± SD. (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Article Snippet: Mouse IL6 Precoated ELISA Kit (Cat:1210602), and Mouse TNF-α Precoated ELISA Kit (Cat:1217202) were purchased from Dakewe, China, and MCP1 ELISA kits were purchased from RenjieBio, China and operated according to the instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Saline

Journal: Journal of Inflammation Research

Article Title: NHWD-870, a Potent BET Inhibitor, Ameliorated Endotoxemia-Induced Hepatic Inflammation via Suppression of BRD4-STAT1 Signaling and Macrophage M1 Polarization

doi: 10.2147/JIR.S598139

Figure Lengend Snippet: NWD-870 improved the survival rate and liver inflammation of mice with endotoxemia. ( A) Schematic diagram of the lethal model of endotoxemia. (B) Survival analysis of mice treated with NHWD-870. Mice were divided into five groups as following: LPS (15mg/kg) +vehicle (n=5), LPS+0.125mg/kg NHWD-870 (n=6), LPS+0.25mg/kg NHWD-870 (n=6), LPS+0.5mg/kg NHWD-870 (n=6), LPS+1.0mg/kg NHWD-870 (n=5). (C) Schematic diagram of NHWD-870 treatment of endotoxemia liver inflammation model. (D and E) Serum levels of ALT and AST. (F) HE staining of liver tissue (Magnification 100×). (G-I) ELISA detection of IL6 ( G ), TNFα ( H ), and MCP1( I ) expression levels in mouse serum. (J-L) RT-qPCR detection of IL6 ( J ), TNFα ( K ), and MCP1 ( L ) mRNA expression levels in mouse liver tissue after NHWD-870 intervention. Each time point of endotoxemia model were divided into 3 groups: vehicle (n=5), NHWD-870 (n=5), JQ1 (n=5). (A and B) showed survival study using 15 mg/kg LPS (lethal dose); (C–L) showed mechanistic study using 7.5 mg/kg LPS (sublethal dose) to enable tissue collection at 12h and 24h. Different doses were selected based on distinct experimental objectives as detailed in Methods. Data are presented as mean ± SD. (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Article Snippet: Mouse IL6 Precoated ELISA Kit (Cat:1210602), and Mouse TNF-α Precoated ELISA Kit (Cat:1217202) were purchased from Dakewe, China, and MCP1 ELISA kits were purchased from RenjieBio, China and operated according to the instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Journal of Inflammation Research

Article Title: NHWD-870, a Potent BET Inhibitor, Ameliorated Endotoxemia-Induced Hepatic Inflammation via Suppression of BRD4-STAT1 Signaling and Macrophage M1 Polarization

doi: 10.2147/JIR.S598139

Figure Lengend Snippet: Effects of NHWD-870 on inflammatory cell infiltration in endotoxemia mice and its effects on various organs. ( A ) IHC detection of F4/80 expression in liver tissues. (scale bar 100μm) ( B ) Statistics of the positive area of F4/80 IHC staining. ( C ) IHC detection of MPO expression in liver tissues. (scale bar 100μm) ( D ) Statistics of the positive area of MPO IHC staining. IHC positive areas are brown-yellow. Each time point of endotoxemia model were divided into 3 groups: vehicle (n=5), NHWD-870 (n=5), JQ1 (n=5). ( E ) HE staining of NHWD-870 treatment on the heart, liver, spleen, kidney, and liver morphology of mice. (Magnification 100×) (F-G): ALT ( F ) and AST ( G ) levels after NHWD-870 intervention. ( H-J ) ELISA detection of IL6 ( H ), TNFα ( I ), and MCP1 ( J ) levels in mouse serum. Vehicle group (n=5); NHWD-870 group (n=5). Data are presented as mean ± standard deviation. (*p<0.05, ***p<0.001, ****p<0.0001).

Article Snippet: Mouse IL6 Precoated ELISA Kit (Cat:1210602), and Mouse TNF-α Precoated ELISA Kit (Cat:1217202) were purchased from Dakewe, China, and MCP1 ELISA kits were purchased from RenjieBio, China and operated according to the instructions.

Techniques: Expressing, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Cellular and Molecular Immunology

Article Title: Hepatocyte-derived LRG1 primes the liver for metastasis and impairs immunotherapy

doi: 10.1038/s41423-026-01408-9

Figure Lengend Snippet: Tumor-associated inflammation promotes the expression of LRG1 in hepatocytes by IL6/STAT3. A The relative expression of Lrg1 in AML12 cells co-cultured with various mouse tumor cell lines or treated with serum (5%) from CRC model mice. n = 3 independent experiments. Data are means ± SD. B Western blot analysis of the expression LRG1 and β-actin in AML12 cells after the indicated treatments. C Cytokine array analyses of serum from CRC orthotopic mice model (day 7, day 14, day 21 and day 28) and from CRC intrasplenic mice model (day 5, day 10, day 15 and day 21). Quantitative real-time PCR analyses of the expression Lrg1 D and ELISA analyses of media of AML12 E treated with vehicle or recombinant IL6/G-CSF/CXCL13/CCL12/TIMP1. n = 3 independent experiments. Data are means ± SD. F, G qRT-PCR and Western blot analyses of LRG1 in AML12 cells treated with cytokine combinations. n = 3 independent experiments. Data are means ± SD. H, I qRT-PCR and western blot analysis of the expression LRG1 and β-actin in AML12 cells treated with indicated serum and anti-IL6 or tocilizumab. n = 3 independent experiments. Data are means ± SD. J ELISA analyses of serum samples for IL6 from sham-group, CRC orthotopic model at day 21(PMN) and CRC intrasplenic model at day 21(Met) in BALB/c. n = 4. Data are means ± SD. K Correlation between serological IL6 and LRG1 levels of CRC patients is shown using Pearson’s correlation analysis. Dots represent individual samples. n = 161. L, M Schematic of the experimental design L . Representative images of liver metastases M in each group ( n = 6 in Lrg1 (+/+)Hep-HTVi-Ctrl group and Lrg1 (+/+)Hep-HTVi-IL6 group, n = 5 in Lrg1 (Δ/Δ)Hep-HTVi-Ctrl group and Lrg1 (Δ/Δ)Hep-HTVi-IL6 group). Scale bars, 1 cm. N Schematic of the experimental design. Representative bioluminescence images and analyses of liver metastases in CRC mouse model treated with anti-IL6 or IgG. O Schematic of the experimental design. P ELISA analyses of samples for IL6 levels from liver interstitial fluid, peripheral blood and tumor interstitial fluid of mice from different groups as indicated. n = 4. Data are means ± SD. Q ELISA analyses of samples for LRG1 levels from liver interstitial fluid and peripheral blood of mice from different groups as indicated. n = 4. Data are means ± SD. R Western blot analyses of LRG1 and β-actin in hepatocyte of mice from different groups as indicated. Statistical significance was determined using two-tailed unpaired Student’s t test A, D–F, H, and N–P

Article Snippet: ELISA kits were used to measure the levels of human LRG1 (Ray Bio), human IL6 (Boster, EK0410), mouse LRG1 (ELK Biotechnology), and mouse IL6 (Boster, EK0411) in cell culture supernatants or serum samples according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Recombinant, Quantitative RT-PCR, Two Tailed Test