Journal: World Journal of Gastroenterology

Article Title: Sirtuin1 attenuates acute liver failure by reducing reactive oxygen species via hypoxia inducible factor 1α

doi: 10.3748/wjg.v28.i17.1798

Figure Lengend Snippet: Hypoxia decreased Sirtuin1 expression leading to the acetylation and activation of hypoxia inducible factor-1α. A-D: Western blotting was performed to measure the levels of Sirtuin1 (Sirt1), Bcl-2 adenovirus E1B-interacting protein 3 (Bnip3), and hypoxia inducible factor (HIF)-1α in L02 cells; E: The representative images of immunofluorescence staining for Acetyl-lysine and HIF-1α; F and G: Equal amounts of protein were subjected to immunoprecipitation with Sirt1 antibody or HIF-1α antibody followed by immunoblotting with antibody against Sirt1, HIF-1α, or acetyl-lysine and effect of Sirt1 overexpression (O/E) was shown. Data shown are means ± standard deviations (SDs) of three separate experiments. a P < 0.05 vs Control group; one-way analysis of variance combined with Bonferroni's post hoc test; the error bars indicate the SDs.

Article Snippet: Human embryonic liver cell line L02 was purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Activation Assay, Western Blot, Immunofluorescence, Staining, Immunoprecipitation, Over Expression, Control

Journal: World Journal of Gastroenterology

Article Title: Sirtuin1 attenuates acute liver failure by reducing reactive oxygen species via hypoxia inducible factor 1α

doi: 10.3748/wjg.v28.i17.1798

Figure Lengend Snippet: The inhibition of Sirtuin1 induced activation of hypoxia inducible factor-1α and subsequently increased hypoxia-induced reactive oxygen species production. A: Western blotting was performed to measure the levels of Sirtuin1 (Sirt1) and hypoxia inducible factor (HIF)-1α in L02 cells; B: Reactive oxygen species (ROS) productions were detected by dihydroethidium (DHE) staining. Representative images of the DHE staining in different groups; C: ROS productions were evaluated by quantification of mean fluorescence intensity in DHE staining; D: Western blotting was performed to measure the levels of Sirt1 and HIF-1α in L02 cells; E and F: ROS productions were detected by DHE staining. Data shown are means ± standard deviations (SDs) of three separate experiments. a P < 0.05 vs Control group; b P < 0.05 vs Lipopolysaccharide (LPS)-treated group; c P < 0.05 vs LPS + Hypoxia-treated group; one-way analysis of variance with Bonferroni's post hoc test; the error bars indicate the SDs.

Article Snippet: Human embryonic liver cell line L02 was purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Inhibition, Activation Assay, Western Blot, Staining, Fluorescence, Control

Journal: World Journal of Gastroenterology

Article Title: Sirtuin1 attenuates acute liver failure by reducing reactive oxygen species via hypoxia inducible factor 1α

doi: 10.3748/wjg.v28.i17.1798

Figure Lengend Snippet: The inhibition of Sirtuin1/peroxisome proliferator-activated receptor alpha signaling pathway increased hypoxia-induced reactive oxygen species production. A: Western blotting was performed to measure the levels of peroxisome proliferator-activated receptor alpha (PPARα) in L02 cells; B: The levels of Sirtuin1 (Sirt1) and hypoxia inducible factor (HIF)-1α in L02 cells; C: Reactive oxygen species productions were detected by dihydroethidium (DHE) staining and evaluated by quantification of mean fluorescence intensity in DHE staining. Data shown are means ± standard deviations (SDs) of three separate experiments. a P < 0.05 vs Control group; b P < 0.05 vs Lipopolysaccharide (LPS)-treated group; c P < 0.05 vs LPS + Hypoxia-treated group; one-way analysis of variance with Bonferroni's post hoc test; the error bars indicate the SDs.

Article Snippet: Human embryonic liver cell line L02 was purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Inhibition, Western Blot, Staining, Fluorescence, Control

Journal: World Journal of Gastroenterology

Article Title: Sirtuin1 attenuates acute liver failure by reducing reactive oxygen species via hypoxia inducible factor 1α

doi: 10.3748/wjg.v28.i17.1798

Figure Lengend Snippet: The inhibition of Sirtuin1/AMP-activated protein kinase signaling pathway increased hypoxia-induced reactive oxygen species production. A: Western blotting was performed to measure the levels of AMP-activated protein kinase (AMPK) and p-AMPK in L02 cells; B: The levels of Sirtuin1 (Sirt1) and hypoxia inducible factor (HIF)-1α in L02 cells; C: Reactive oxygen species (ROS) productions were detected by dihydroethidium (DHE) staining and evaluated by quantification of mean fluorescence intensity in DHE staining. Data shown are means ± standard deviations (SDs) of three separate experiments. a P < 0.05 vs Control group; b P < 0.05 vs Lipopolysaccharide (LPS)-treated group; c P < 0.05 vs LPS + Hypoxia-treated group; one-way analysis of variance combined with Bonferroni's post hoc test; the error bars indicate the SDs.

Article Snippet: Human embryonic liver cell line L02 was purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Inhibition, Western Blot, Staining, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: The protective effects of Antrodia cinnamomea extract (ACE) against irradiation in human normal liver CL48 cells. ( A ) Cytotoxicity of irradiation in CL48. Cells were treated with different dosages (0, 5, 10, 15, and 20 Gy) of irradiation and counted by hemocytometer at intervals as indicated. ( B ) Determination of the optimal ACE dosages. Cells were treated with different concentrations of ACE for 24 h or 48 h, followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cell survival. For the control group, the same concentrations of ethanol were used for each ACE treatment experiment. ( C ) The protective effect of ACE against irradiation. Cells were pre-treated with different concentrations of ACE for 16 h followed by various dosages of irradiation; they were evaluated by MTT assay 48 h later. Cell survival was evaluated by MTT assay. Results were obtained from three independent experiments; each experiment was done in triplicate. **, p < 0.01, as compared with the control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, MTT Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Protective effects of ACE against irradiation-induced apoptosis. CL48 cells were pre-treated with various concentrations of ACE for 16 h followed by irradiation at 15 Gy. Cells were harvested 48 h later for determination of caspase-3 activity ( A ), caspase-8 activity ( B ), and caspase-9 activity ( C ). The fold differences of the three caspase activities relative to individual control groups were also presented ( D ). For apoptosis analysis, cells were harvested 30 h after treatment followed by staining with PI and Annexin V ( E ). Results are representative of three independent experiments. The statistical analysis of the percentages of early plus late apoptotic cells ( F ). *, p < 0.05; **, p < 0.01, as compared with the irradiated ACE 0 μg/mL control group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Activity Assay, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Reactive oxygen species (ROS) scavenging activity of ACE in CL48 cells. After being treated and harvested as described in the Materials and Methods section, cells were analyzed by flowcytometry after incubation with 2′,7′-dichlorofluorescin diacetate (DCFH-DA) for 15 min. ( A ) Control cells without treatment, ( B ) IR treated only, ( C ) H 2 O 2 treated only, ( D ) pre-treated with NAC before IR, ( E ) pre-treated with ACE 40 μg/mL before IR, ( F ) pre-treated with ACE 80 μg/mL before IR. ( G ) Histograms of B, E, and F were combined to show the dose-dependent ROS scavenging activity of ACE. ( H ) Fold change of control fluorescence intensity by 40 μg/mL and 80 μg/mL of ACE, respectively.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Activity Assay, Incubation, Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Redox-related enzymes expression and activity profiles of CL48 hepatocytes after IR and/or ACE treatments. With or without ACE pre-treatment for 16 h, cells were irradiated with a dose of 15 Gy and harvested 24 h later for RT-PCR ( A ), Western blot ( B ), total superoxide dismutase (SOD) activity ( C ), and glutathione peroxidase (GPx) activity ( D ) analyses. Results were obtained from three independent experiments, each experiment was done in triplicate. **, p < 0.01, as compared with the control IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Activity Assay, Irradiation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

doi: 10.3390/ijms20040846

Figure Lengend Snippet: Enhancement of the irradiation-induced nuclear factor erythroid-2-related factor (Nrf2) expression and nuclear translocation by ACE treatment. With or without ACE pre-treatment for 16 h, CL48 cells were irradiated with a dose of 15 Gy and harvested 24 h later for Western blot ( A ) and immunofluorescence staining ( B , left) of Nrf2. The slides were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (300 nM in PBS) ( B , middle) and merged images ( B , right). ( C ) The nuclear Nrf2 positivity rates were plotted. Results were obtained from random fields of three independent experiments for each group. *, p < 0.05; **, p < 0.01, as compared with the IR only group.

Article Snippet: Normal human embryonic liver cell line CL48 cells (American Type Culture Collection) were cultured in high glucose DMEM with 10% fetal bovine serum (FBS), 0.1 mg/mL of streptomycin, and 100 units/mL of penicillin, at 37 °C in a humidified atmosphere containing 5% CO 2 .

Techniques: Irradiation, Expressing, Translocation Assay, Western Blot, Immunofluorescence, Staining