[Homocysteine promotes human hepatocyte autophagy through activating the activating molecule in beclin-1-regulated autophage (AMBRA1)].

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  • Additional Information
    • Source:
      Publisher: Xi bao yu fen zi mian yi xue za zhi bian ji bu Country of Publication: China NLM ID: 101139110 Publication Model: Print Cited Medium: Print ISSN: 1007-8738 (Print) Linking ISSN: 10078738 NLM ISO Abbreviation: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi Subsets: MEDLINE
    • Publication Information:
      Original Publication: Xiʹan : Xi bao yu fen zi mian yi xue za zhi bian ji bu
    • Subject Terms:
      Adaptor Proteins, Signal Transducing*/genetics ; Autophagy* ; Homocysteine*/pharmacology; Autophagosomes/metabolism ; Hepatocytes/metabolism ; Humans
    • Abstract:
      Objective To investigate the role of activating molecule in beclin-1-regulated autophage (AMBRA1) in homocysteine (Hcy)-induced hepatocytes autophagy. Methods Hepatocytes were cultured in vitro and divided into control group (0 μmol/L Hcy) and Hcy treatment group (100 μmol/L Hcy). Western blotting was used to detect the expression of microtubule-associated protein 1 light chain 3B (LC3BII, LC3BI); hepatocytes were treated with 0, 25, 50, 100 μmol/L chloroquine (CQ), CCK-8 assay was used to detect the inhibitory effect of CQ on hepatocyte proliferation and Western blotting was performed to detect the expression of LC3B and AMBRA1; After hepatocytes were transfected with AMBRA1 small interfering RNA, real-time fluorescent quantitative PCR and Western blotting were used to detect the interference efficiency of AMBRA1 expression; After the transfected hepatocytes were treated with Hcy, the expression of LC3B was detected by Western blot analysis. mRFP-GFP-LC3 adenovirus was transfected with hepatocytes and the autophagy flow was observed by laser scanning confocal microscopy. Results Compared with the control group, the ratio of LC3BII/LC3BIincreased in the Hcy treatment group; the inhibition rate of 50 μmol/L CQ on hepatocyte proliferation was close to 50%; compared with the control group, the ratio of LC3BII/LC3BI and the expression of AMBRA1 increased significantly in the Hcy group , and the ratio of LC3BII/LC3BI and the expression of AMBRA1 in the Hcy combined with CQ group were significantly lower than those in the Hcy group; the ratio of LC3BII/LC3BI decreased after knocking down AMBRA1; compared with the control group, the autophagosomes and autophagolysosomes increased in Hcy group and decreased after knocking down AMBRA1. Conclusion Hcy can promote hepatocyte autophagy by activating AMBRA1.
    • Accession Number:
      0 (AMBRA1 protein, human)
      0 (Adaptor Proteins, Signal Transducing)
      0LVT1QZ0BA (Homocysteine)
    • Publication Date:
      Date Created: 20220909 Date Completed: 20220912 Latest Revision: 20220920
    • Publication Date:
      20221213
    • Accession Number:
      36082712