中文English
ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R
Volume 39 Issue 12
Dec.  2023
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Article Contents

Mechanism of microRNA-223-3p inhibiting hepatic stellate cell activation by targeting microtubule-associated protein 1B

DOI: 10.3969/j.issn.1001-5256.2023.12.015
Research funding:

National Natural Science Fundation of China (8200033704);

National Natural Science Fundation of China (81770565)

More Information
  • Corresponding author: LI Ning, lining_hs@fudan.edu.cn (ORCID: 0000-0002-4375-2857)
  • Received Date: 2022-12-09
  • Accepted Date: 2023-05-19
  • Published Date: 2023-12-12
  •   Objective  To investigate the effect of microRNA-223-3p (miR-223-3p) on hepatic stellate cell (HSC) activation and its mechanism.  Methods  Human HSC LX2 cells were selected for the study, and LX2 cells were stimulated by TGF-β to establish a model of HSC activation; quantitative real-time PCR was used to measure the change in the expression level of miR-223-3p during HSC activation. After LX2 cells were transfected with miR-223-3p mimic, quantitative real-time PCR, Western blot, and immunofluorescence assay were used to clarify the regulatory effect of miR-223-3p on HSC activation, and dual-luciferase reporter assay was used to verify the association between miR-223-3p and the target gene MAP1B. After LX2 cells were transfected with MAP1B siRNA, Western blot was used to clarify the influence of inhibiting MAP1B expression on HSC activation; after LX2 cells were transfected with miR-223-3p, quantitative real-time PCR and Western blot were used to verify the regulatory effect of miR-223-3p on MAP1B. The independent-samples t test was used for comparison of continuous data between two groups.  Results  HSC in the activated state had a significant reduction in the expression level of miR-223-3p compared with those in the resting state (t=9.12, P<0.001). Overexpression of miR-223-3p inhibited the mRNA and protein expression levels of the markers for HSC activation alpha-smooth muscle actin and collagen type Ⅰ (mRNA expression: t=8.35 and 12.23, both P<0.01; protein expression: t=16.24 and 20.90, both P<0.001). The dual-luciferase reporter assay confirmed that MAP1B was a potential target gene of miR-223-3p. Compared with the control group, LX2 cells with miR-223-3p overexpression had significant reductions in the mRNA and protein expression levels of MAP1B (mRNA expression: t=5.95, P<0.01; protein expression: t=11.12, P<0.001).  Conclusion  This study shows that miR-223-3p can inhibit HSC activation by targeting MAP1B.

     

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