Journal: Journal of Clinical and Translational Hepatology

Article Title: Investigation of HO-1 Regulation of Liver Fibrosis Related to Nonalcoholic Fatty Liver Disease Through the SIRT1/TGF-ß/Smad3 Pathway

doi: 10.14218/JCTH.2024.00481

Figure Lengend Snippet: (A) liver /body weight. (B) Body weight. Serum (C) ALT and (D) AST in each group. (E) Images of H&E (×200 magnification) staining of representative liver sections and (F) the NAFLD activity score. (G) Images of Masson (×200 magnification) staining of representative liver sections and (H) the Metavir score. (I) Images of Sirius (×200 magnification) red staining of representative liver sections and (J) Relative collagen content (%). Values are the mean ± SD (n = 6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. MCD, methionine-and-choline-deficient diet; NAFLD, nonalcoholic fatty liver disease; H&E, hematoxylin-eosin. ALT, alanine aminotransferase; AST, aspartate transaminase; Znpp, zincprotoporphyria.

Article Snippet: The mice were randomly divided into four groups (n = 6) and received group-specific diets for eight weeks: (1) Control group: Mice were fed a normal diet; (2) Methionine- and choline-deficient (MCD) group: Mice were fed an MCD diet (Research Diets, Inc., NJ, New Brunswick, USA); (3) Hemin group: Mice were fed an MCD diet and treated with the HO-1 chemical inducer Hemin three times per week; (4) Zinc protoporphyrin (Znpp) group: Mice were fed an MCD diet and treated with the HO-1 inhibitor Znpp three times per week.

Techniques: Staining, Activity Assay

Journal: Journal of Clinical and Translational Hepatology

Article Title: Investigation of HO-1 Regulation of Liver Fibrosis Related to Nonalcoholic Fatty Liver Disease Through the SIRT1/TGF-ß/Smad3 Pathway

doi: 10.14218/JCTH.2024.00481

Figure Lengend Snippet: (A–F) The expression levels of HO-1, SIRT1, TGF-β, Smad3, Collagen1 and α-SMA were measured by RT-qPCR. (G) Immunofluorescence (×200 magnification) double staining between of HO-1 and SIRT1 in liver tissue. (H–K) Immunofluorescence double staining semi-quantitative analysis. GAPDH served as the loading control. Data are presented as representative results of three independent experiments. Values are the mean ± SD (n = 6 per group). *** p < 0.001, **** p < 0.0001. MCD, methionine-and-choline-deficient diet; HO-1, heme oxygenase 1; SIRT1, Sirtuin 1; TGF-β, transforming growth factor beta; α-SMA, alpha-smooth muscle actin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Znpp, zincprotoporphyria.

Article Snippet: The mice were randomly divided into four groups (n = 6) and received group-specific diets for eight weeks: (1) Control group: Mice were fed a normal diet; (2) Methionine- and choline-deficient (MCD) group: Mice were fed an MCD diet (Research Diets, Inc., NJ, New Brunswick, USA); (3) Hemin group: Mice were fed an MCD diet and treated with the HO-1 chemical inducer Hemin three times per week; (4) Zinc protoporphyrin (Znpp) group: Mice were fed an MCD diet and treated with the HO-1 inhibitor Znpp three times per week.

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Double Staining, Control

Journal: Journal of Clinical and Translational Hepatology

Article Title: Investigation of HO-1 Regulation of Liver Fibrosis Related to Nonalcoholic Fatty Liver Disease Through the SIRT1/TGF-ß/Smad3 Pathway

doi: 10.14218/JCTH.2024.00481

Figure Lengend Snippet: (A, C–G) Western blot analysis of HO-1, SIRT1, TGF-β, P-Smad2/3, and Smad2/3 in liver tissue. (B, H, I) Western blot analysis of α-SMA and Collagen1 in liver tissue. Values are the mean ± SD (n = 6 per group). ** p < 0.01, *** p < 0.001, **** p < 0.0001. MCD, methionine-and-choline-deficient diet; HO-1, heme oxygenase 1; SIRT1, Sirtuin 1; TGF-β, transforming growth factor beta; α-SMA, alpha-smooth muscle actin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Znpp, zincprotoporphyria.

Article Snippet: The mice were randomly divided into four groups (n = 6) and received group-specific diets for eight weeks: (1) Control group: Mice were fed a normal diet; (2) Methionine- and choline-deficient (MCD) group: Mice were fed an MCD diet (Research Diets, Inc., NJ, New Brunswick, USA); (3) Hemin group: Mice were fed an MCD diet and treated with the HO-1 chemical inducer Hemin three times per week; (4) Zinc protoporphyrin (Znpp) group: Mice were fed an MCD diet and treated with the HO-1 inhibitor Znpp three times per week.

Techniques: Western Blot

Journal: Journal of Clinical and Translational Hepatology

Article Title: Investigation of HO-1 Regulation of Liver Fibrosis Related to Nonalcoholic Fatty Liver Disease Through the SIRT1/TGF-ß/Smad3 Pathway

doi: 10.14218/JCTH.2024.00481

Figure Lengend Snippet: (A, B) Western blot analysis of HO-1 overexpression and silencing in LX2 cells. (C, D) Western blot analysis of LX2 cells treated with SIRT1 activators and inhibitors. (E–I) Western blot analysis of SIRT1, TGF-β, P-Smad2/3, Smad2/3 in LX2 cells. (J–L) Western blot analysis of α-SMA and Collagen I in LX2 cells. Values are the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. MCD, methionine-and-choline-deficient diet; HO-1, heme oxygenase 1; SIRT1, Sirtuin 1; TGF-β, transforming growth factor beta; α-SMA, alpha-smooth muscle actin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; OE HO-1, overexpressing HO-1; SiHO-1, small interfering RNA silencing of HO-1.

Article Snippet: The mice were randomly divided into four groups (n = 6) and received group-specific diets for eight weeks: (1) Control group: Mice were fed a normal diet; (2) Methionine- and choline-deficient (MCD) group: Mice were fed an MCD diet (Research Diets, Inc., NJ, New Brunswick, USA); (3) Hemin group: Mice were fed an MCD diet and treated with the HO-1 chemical inducer Hemin three times per week; (4) Zinc protoporphyrin (Znpp) group: Mice were fed an MCD diet and treated with the HO-1 inhibitor Znpp three times per week.

Techniques: Western Blot, Over Expression, Small Interfering RNA

Journal: Lipids in Health and Disease

Article Title: Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

doi: 10.1186/s12944-019-1114-4

Figure Lengend Snippet: Body weight change, fat pad weight, liver weight, and hepatic triglyceride (TG) and malondialdehyde (MDA) levels in mice fed a methionine-choline-deficient (MCD) or control diet and housed in two different animal facilities (WT1, WT2). a Body weight (BW) at the end of the study relative to the initial weight in %. b Epididymal (Epi) fat pad weight normalized to BW. c Liver weight normalized to BW. d H&E stained liver of WT mice fed a control diet. Small lipid droplets appear in the liver of WT2 mice and are visible at higher magnification (second row). Large lipid droplets in the liver of MCD diet fed mice (third and fourth row). e Oil Red O stained liver. f Hepatic TG concentrations. g Hepatic MDA levels. Data of 5 to 8 animals per group are shown. * p < 0.05, ** p < 0.01

Article Snippet: Fifteen mice (14 weeks old - 8 WT1 and 7 WT2) were fed the MCD diet (E15653–94, Ssniff, Soest, Germany) and eleven mice (14 weeks old - 6 WT1 and 5 WT2 animals) the respective control chow (E15654–04, Ssniff, Soest, Germany) for 2 weeks.

Techniques: Control, Staining

Journal: Lipids in Health and Disease

Article Title: Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

doi: 10.1186/s12944-019-1114-4

Figure Lengend Snippet: Hepatic expression of inflammatory and profibrotic genes in mice fed a methionine-choline-deficient (MCD) or control diet and housed in two different animal facilities (WT1, WT2). a Hepatic F4/80 mRNA. b Hepatic CD68 mRNA. c Hepatic TNF mRNA. d Hepatic IL-6 mRNA. e Hepatic chemerin mRNA. f Hepatic chemerin protein. g Quantification of hepatic chemerin protein. h Hepatic TGF beta mRNA. i Hepatic alpha SMA mRNA. Data of 5 to 8 animals per group are shown. * p < 0.05, ** p < 0.01

Article Snippet: Fifteen mice (14 weeks old - 8 WT1 and 7 WT2) were fed the MCD diet (E15653–94, Ssniff, Soest, Germany) and eleven mice (14 weeks old - 6 WT1 and 5 WT2 animals) the respective control chow (E15654–04, Ssniff, Soest, Germany) for 2 weeks.

Techniques: Expressing, Control

Journal: Lipids in Health and Disease

Article Title: Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

doi: 10.1186/s12944-019-1114-4

Figure Lengend Snippet: Cholesteryl ester (CE), free cholesterol (FC), sphingomyelin (SM) and ceramide (Cer) in the liver of mice fed a methionine-choline-deficient (MCD) or control diet and housed in two different animal facilities (WT1, WT2). Concentrations are given as nmol/mg wet weight. a CE b FC c CE 22:6 d Total SM e SM 40:1 f SM 34:1 g Total Cer h Cer d18:1/16:0 and ( i ) Cer d18:1/22:0. Data of 5 to 8 animals per group are shown. * p < 0.05, ** p < 0.01

Article Snippet: Fifteen mice (14 weeks old - 8 WT1 and 7 WT2) were fed the MCD diet (E15653–94, Ssniff, Soest, Germany) and eleven mice (14 weeks old - 6 WT1 and 5 WT2 animals) the respective control chow (E15654–04, Ssniff, Soest, Germany) for 2 weeks.

Techniques: Control

Journal: Lipids in Health and Disease

Article Title: Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

doi: 10.1186/s12944-019-1114-4

Figure Lengend Snippet: Lysophosphatidylcholine (LPC), phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in the liver of mice fed a methionine-choline-deficient (MCD) or control diet and housed in two different animal facilities (WT1, WT2). Concentrations are given as nmol/mg wet weight. a LPC 20:3 b LPC 16:0 c saturated (Sat) LPC d monounsaturated (MUFA) PC e PC 32:1 f PC 30:0 g PC to PE ratio h PE 36:5 and i PE 40:6. Data of 5 to 8 animals per group are shown. * p < 0.05, ** p < 0.01

Article Snippet: Fifteen mice (14 weeks old - 8 WT1 and 7 WT2) were fed the MCD diet (E15653–94, Ssniff, Soest, Germany) and eleven mice (14 weeks old - 6 WT1 and 5 WT2 animals) the respective control chow (E15654–04, Ssniff, Soest, Germany) for 2 weeks.

Techniques: Control

Journal: Lipids in Health and Disease

Article Title: Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

doi: 10.1186/s12944-019-1114-4

Figure Lengend Snippet: Phosphatidylserine (PS) and phosphatidylinositol (PI) in the liver of mice fed a methionine-choline-deficient (MCD) or control diet and housed in two different animal facilities (WT1, WT2). Concentrations are given as nmol/mg wet weight. a PS 40:5 b Monounsaturated (MUFA) PS c PS 36:4 d PI 40:4 e PI 34:2 and f PI 40:5. Data of 5 to 8 animals per group are shown. * p < 0.05, ** p < 0.01

Article Snippet: Fifteen mice (14 weeks old - 8 WT1 and 7 WT2) were fed the MCD diet (E15653–94, Ssniff, Soest, Germany) and eleven mice (14 weeks old - 6 WT1 and 5 WT2 animals) the respective control chow (E15654–04, Ssniff, Soest, Germany) for 2 weeks.

Techniques: Control

Journal: Journal of biochemical and molecular toxicology

Article Title: Shc inhibitor idebenone ameliorates liver injury and fibrosis in dietary NASH in mice

doi: 10.1002/jbt.22876

Figure Lengend Snippet: Shc expression and activation in PBMC and liver in MCD model. Six-week-old male C57B6J mice were fed with an MCD diet for 6 weeks with idebenone 10 or 40 mg/kg oral daily starting at Day 1 of the diet representing a preventive treatment. Shc phosphorylation and activation in PBMC (A) and liver (B) were analyzed with either Jess capillary Western blot or conventional Western blot analysis. The image signal was quantified with the Compass software. Phosphorylation and expression of p52Shc increased in PBMC in the MCD group and dramatically reduced by idebenone on both 10 and 40 mg/kg. In the liver, Shc expression had a trend of reduction by idebenone, but less extent compared to PBMC. Representative images are shown; mean ± SD, N = 4. Ctrl, control; Ide, idebenone; MCD, methionine-choline deficient; PBMC, peripheral blood mononuclear cell. *p < 0.05

Article Snippet: For the MCD model, the 8-week old mice were given either a control diet supplemented with methionine and choline or an MCD diet (Bio-Serv) for 6 weeks.

Techniques: Expressing, Activation Assay, Western Blot, Software

Journal: Journal of biochemical and molecular toxicology

Article Title: Shc inhibitor idebenone ameliorates liver injury and fibrosis in dietary NASH in mice

doi: 10.1002/jbt.22876

Figure Lengend Snippet: Idebenone protects mice from MCD diet-induced liver injury. The mice were on an MCD diet with preventive idebenone dosing for 6 weeks. Serological study showed that idebenone 40 mg/kg decreased the ALT level significantly (A). AST had a trend of decrease at both 10 and 40 mg/kg (B). Real-time qPCR on liver tissues (C) showed that the idebenone reduced the transcripts of fibrogenic genes (Col1a1, αSMA, TGFβ, MMP2, and Timp1) significantly. The reduction of Col1a1 showed a dose-dependent trend. Hydroxyproline assay (D) showed a lower amount of collagen content detected in both dosage groups. Picrosirius red staining and H&E (E) showed improved fibrosis and histology in the treated mice. Arrows indicate the patches of inflammatory cells; mean ± SD, N = 8–10. ALT, aminotransferase; AST, aspartate aminotransferase; Col1a1, collagen type I alpha 1 chain; H&E, hematoxylin and eosin; Ide, idebenone; MCD, methionine-choline deficient; MMP2, matrix metalloproteinase 2; qPCR, quantitative polymerase chain reaction; TGFβ, transforming growth factor-β; αSMA, α-smooth muscle actin. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: For the MCD model, the 8-week old mice were given either a control diet supplemented with methionine and choline or an MCD diet (Bio-Serv) for 6 weeks.

Techniques: Hydroxyproline Assay, Staining, Real-time Polymerase Chain Reaction