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Downregulation of miR-20b-5p Contributes to the Progression of Liver Fibrosis via the STAT3 Signaling Pathway In Vivo and In Vitro

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Abstract

Background

Activated hepatic stellate cells (HSCs) are primarily involved in liver fibrosis and portal hypertension (PHT). We aimed to investigate the effect of miR-20b-5p on HSCs, liver fibrosis, and PHT.

Methods

MiR-20b-5p expression in HSCs and in mouse liver fibrosis was determined by qPCR. Further, the effects of miR-20b-5p mimic on HSCs migration, proliferation, and apoptosis were investigated in vitro. A dual-luciferase reporter assay was performed to confirm the direct interaction between miR-20b-5p and STAT3. In vivo, mouse liver fibrosis was established by common bile duct ligation and intervened by agomiR-20b-5p. Sirius red staining and hydroxyproline content were used to evaluate collagen deposition. The α-SMA expression in the liver was detected by IHC and Western blotting. The STAT3 signaling pathway and its downregulated cytokines as well as portal pressure and angiogenesis were explored.

Results

MiR-20b-5p was significantly downregulated during HSCs activation and in mouse liver fibrosis. The functional analyses demonstrated that miR-20b-5p inhibited cell proliferation, activation, and promoted apoptosis in HSCs in vitro. Moreover, miR-20b-5p regulated STAT3 expression by binding to the 3'UTR of its miRNA directly. Overexpression of miR-20b-5p facilitated HSC activation and proliferation by inhibiting the STAT3 signaling pathway. MiR-20b-5p overexpression suppressed the STAT3 and its downstream cytokines and ameliorated liver fibrosis in mice. The intra- and inter-hepatic angiogenesis were also effectively inhibited. The inhibition of liver fibrosis and neoangiogenesis contributed to the decrease of portal pressure.

Conclusions

MiR-20b-5p plays an important role in the fibrosis and angiogenesis of liver fibrosis by targeting the STAT3 signaling pathway.

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Funding

The study was supported by the National Natural Science Foundation of China (No. 81700533), the Key Research and Development Program of Shaanxi Province, China (Nos. 2017SF-116, 2015JM8420, 2015KTCL03-05), and the Innovation and Development Foundation of Tangdu Hospital (Nos. 2017LCYJ003, 2018JSYJ010, 2019LCYJ005).

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Author notes

  1. Ling Lv and Dong Wang contributed equally to the article.

Authors and Affiliations

  1. Department of Disease Control and Prevention, Tangdu Hospital, Fourth Military Medical University, 569 Xin Si Road, Xi’an, 710038, China

    Ling Lv

  2. Department of General Surgery, Tangdu Hospital, Fourth Military Medical University, Xi’an, China

    Dong Wang, Jikai Yin, Tao Yang, Bo Huang, Yanlong Cao & Jianguo Lu

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  1. Ling Lv
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  2. Dong Wang
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  3. Jikai Yin
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  4. Tao Yang
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Correspondence to Dong Wang.

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Lv, L., Wang, D., Yin, J. et al. Downregulation of miR-20b-5p Contributes to the Progression of Liver Fibrosis via the STAT3 Signaling Pathway In Vivo and In Vitro. Dig Dis Sci 68, 487–496 (2023). https://doi.org/10.1007/s10620-022-07660-z

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