Vitamin D and NAFLD Quiz
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Questions and Answers

What is the purpose of CCK-8 solution in the experiment?

  • To stain lipid droplets in cells
  • To dehydrate liver tissue slices
  • To isolate DNA from fecal samples
  • To measure the optical density at 450 nm (correct)
  • Which staining method is employed for detecting lipid droplets in the study?

  • Immunofluorescence (IF) staining
  • DAPI staining
  • Oil red O staining (correct)
  • Hematoxylin-eosin (H&E) staining
  • Which step is performed after blocking with goat serum during the immunofluorescence staining procedure?

  • Incubation with DAPI for counterstaining
  • Staining with hematoxylin
  • Embedding in paraffin
  • Incubation with NLRP3 antibodies (correct)
  • What is the magnification of the fluorescence microscope used for imaging?

    <p>400x</p> Signup and view all the answers

    What method is used to analyze gut microbiota changes in the study?

    <p>16s ribosomal RNA sequencing</p> Signup and view all the answers

    What role does VD play in relation to HFD-induced hepatic injury?

    <p>It protects against HFD-induced hepatic injury.</p> Signup and view all the answers

    Which of the following components does pyroptosis involve?

    <p>Activation of NLRP3 inflammasome.</p> Signup and view all the answers

    How does GSDMD-N fragment overexpression affect the protective effects of VD?

    <p>It suppresses the protective effects of VD.</p> Signup and view all the answers

    What was observed regarding gut microbiota due to VD treatment?

    <p>Increase in Lactobacillus and decrease in specific harmful bacteria.</p> Signup and view all the answers

    What is one consequence of NAFLD as mentioned in the content?

    <p>Increased risk of hepatocellular carcinoma.</p> Signup and view all the answers

    Study Notes

    Vitamin D and NAFLD

    • Vitamin D (VD) attenuated hepatic injury and lipid accumulation, reduced NLRP3 inflammasome activation and pyroptosis both in vivo (mice) and in vitro (BRL-3A cells).
    • GSDMD-N fragment overexpression suppressed the protective effect of VD on cell viability and lipid accumulation.
    • VD restored gut microbiota dysbiosis by increasing the relative abundance of Lactobacillus and reducing that of Acetatifactor, Oscillibacter and Flavonifractor.

    NAFLD

    • Non-alcoholic fatty liver disease (NAFLD) is a spectrum of liver conditions from fatty liver to cirrhosis and even hepatocellular carcinoma.
    • NAFLD affects 25% of the adult population globally.

    Pyroptosis

    • Pyroptosis is an inflammatory form of programmed cell death.
    • The canonical pathway is initiated by inflammasomes, which activate caspase-1.
    • Caspase-1 cleaves pro-IL-1β and pro-IL-18 into their active forms (mature IL-1β and IL-18).
    • Caspase-1 also cleaves gasdermin D (GSDMD) into a N-terminal and C-terminal fragment.

    Gut Microbiota

    • HFD consumption significantly decreased the relative abundance of Bacteroidetes while VD administration significantly decreased the relative abundance of Firmicutes and increased the relative abundance of Bacteroidetes.
    • VD effectively relieved HFD-induced gut microbiota dysbiosis.

    Experiment Results

    • In vivo: VD administration restored the HFD-triggered production of fat vacuoles and disordered structure of hepatic lobules.
    • In vitro: VD significantly decreased the expression of NLRP3, ASC, pro-caspase-1, cleaved-caspase-1, pro-IL-1β, IL-1β and GSDMD-N in BRL-3A cells.
    • VD significantly decreased TG content and lipid accumulation in BRL-3A cells.
    • GSDMD-N fragment overexpression reversed the VD-induced decreases in the expression of GSDMD-N, NLRP3, ASC, cleaved-caspase-1, pro-IL-1β, IL-1β and GSDMD-N, indicating that VD might attenuate NAFLD through inhibiting pyroptosis.
    • GSDMD-N fragment overexpression significantly reversed the VD-induced decreases in loss of cell viability, indicating that VD promoted the viability of the BRL-3A cells stimulated by PA and LPS via the regulation of pyroptosis.
    • GSDMD-N fragment overexpression significantly reversed the VD-induced reduction in TG contents and accumulation of lipid droplets.

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    Description

    Test your knowledge on the relationship between Vitamin D and Non-Alcoholic Fatty Liver Disease (NAFLD). This quiz will cover the effects of Vitamin D on hepatic injury, lipid accumulation, and the mechanisms of pyroptosis in the context of NAFLD. Explore how gut microbiota plays a role in these processes as well.

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