Journal: Frontiers in Medicine

Article Title: Myricetin Modulates Macrophage Polarization and Mitigates Liver Inflammation and Fibrosis in a Murine Model of Nonalcoholic Steatohepatitis

doi: 10.3389/fmed.2020.00071

Figure Lengend Snippet: Experimental study design. (A) The chemical structure of myricetin (3,3',4',5,5',7-hexahydroxyflavone). (B) Experimental protocol for assessment the preventive effect of myricetin (Myr) on the development of NASH and fibrosis in mice fed the choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD). Group A: control mice fed normal chow diet (NCD) and treated with the vehicle (Veh, 0.5% CMC-Na). Group B or C: CDAHFD-induced-NASH mice were randomly assigned to a treatment of Myr (100 mg/kg) or Veh by daily orally administration for 8 weeks.

Article Snippet: Mice were maintained in a temperature-controlled room (23 ± 3, 55 ± 10% humidity) with a 12-h light-dark cycle and had unrestricted to normal chow diet (NCD) or a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) consisting of 60% kcal fat and 0.1% methionine (Research Diets, Inc., New Brunswick, NJ, USA) ( , ).

Techniques: Control

Journal: Frontiers in Medicine

Article Title: Myricetin Modulates Macrophage Polarization and Mitigates Liver Inflammation and Fibrosis in a Murine Model of Nonalcoholic Steatohepatitis

doi: 10.3389/fmed.2020.00071

Figure Lengend Snippet: The impact of myricetin treatment on body weight, liver weight, and serum enzymes at 8 weeks in mice fed NCD or CDAHFD.

Article Snippet: Mice were maintained in a temperature-controlled room (23 ± 3, 55 ± 10% humidity) with a 12-h light-dark cycle and had unrestricted to normal chow diet (NCD) or a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) consisting of 60% kcal fat and 0.1% methionine (Research Diets, Inc., New Brunswick, NJ, USA) ( , ).

Techniques:

Journal: Frontiers in Medicine

Article Title: Myricetin Modulates Macrophage Polarization and Mitigates Liver Inflammation and Fibrosis in a Murine Model of Nonalcoholic Steatohepatitis

doi: 10.3389/fmed.2020.00071

Figure Lengend Snippet: Myricetin inhibited liver fibrosis and HSC activation in NASH mice. (A) Masson's trichrome staining of liver sections in mice from fed an NCD or CDAHFD with myricetin (Myr) or Vehicle (Veh). Upper panel (original magnification, ×100) and lower panel (Original magnification, ×200). (B) Immunohistochemical (IHC) images of liver sections stained for α-smooth muscle actin (α-SMA). Bottom row contains images enlarged from the boxed area in the corresponding panel in the top row. Original magnification, ×100 (upper panel) and ×200 (lower panel). (C) Quantitative analysis of Masson's trichrome positive area. (D) Quantification of the α-SMA-positive area. (E) Western blotting analysis of α-SMA protein in livers; results were normalized relative to expression of GAPDH. (F) Transcript levels of fibrotic markers (α-SMA, Col1α1, CTGF, TIMP-1, and MMP-9) were measured by quantitative RT-PCR in whole liver samples ( n = 5). Results were normalized to β-actin mRNA and expressed as folds change compared to NCD-fed mice. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; “NS” indicates not significant.

Article Snippet: Mice were maintained in a temperature-controlled room (23 ± 3, 55 ± 10% humidity) with a 12-h light-dark cycle and had unrestricted to normal chow diet (NCD) or a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) consisting of 60% kcal fat and 0.1% methionine (Research Diets, Inc., New Brunswick, NJ, USA) ( , ).

Techniques: Activation Assay, Staining, Immunohistochemical staining, Western Blot, Expressing, Quantitative RT-PCR

Journal: Frontiers in Medicine

Article Title: Myricetin Modulates Macrophage Polarization and Mitigates Liver Inflammation and Fibrosis in a Murine Model of Nonalcoholic Steatohepatitis

doi: 10.3389/fmed.2020.00071

Figure Lengend Snippet: Myricetin treatment inhibits the TREM-1-TLR2/4-MyD88 signaling in the liver of CDAHFD-treated mice and in LPS-stimulated RAM267.4 cells. (A) Immunofluorescent double staining in liver sections from each group. Liver sections were double stained for TREM-1 (green) and F4/80 (Red) macrophages. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Original magnification: ×200; Scale bar = 50 μm. (B) Quantification of TREM-1 and F4/80 double-positive cells. (C) Western blotting and quantification of TREM-1 expression in lysed liver tissues, with results relative to the expression of GAPDH ( n = 3). (D) Hepatic mRNA expression of TREM-1, TLR2, TLR4, and MyD88 was measured by quantitative RT-PCR. Results are shown as fold change compared with NCD-fed mice and β-actin served as loading control ( n = 5). (E) The mRNA expression of TREM-1, TLR2, TLR4, and MyD88 in RAW264.7 cells were measured by quantitative RT-PCR. The mRNA levels were normalized to β-actin mRNA levels and presented as fold stimulation (mean ± SEM) vs. DMSO. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; “NS” indicates not significant.

Article Snippet: Mice were maintained in a temperature-controlled room (23 ± 3, 55 ± 10% humidity) with a 12-h light-dark cycle and had unrestricted to normal chow diet (NCD) or a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) consisting of 60% kcal fat and 0.1% methionine (Research Diets, Inc., New Brunswick, NJ, USA) ( , ).

Techniques: Double Staining, Staining, Western Blot, Expressing, Quantitative RT-PCR, Control