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MicroRNA-34b-5p binds enhancer of zeste 2 to inhibit milk fat globule-EGF factor 8 expression, affecting liver fibrosis

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Abstract

Activated hepatic stellate cells (HSCs) are considered the major drivers in the process of hepatic fibrosis. This study intends to explore the mechanism underlying microRNA (miR)-34b-5p effects over liver fibrosis through the enhancer of zeste 2 (EZH2)/milk fat globule-EGF factor 8 (MFGE8) axis in HSCs. A liver fibrosis model was generated by carbon tetrachloride (CCl4) in C57BL/6 J mice and subjected to histological examinations and detection of HSC activation and miR-34b-5p/EZH2/MFGE8 expression. Primary HSCs were treated with transforming growth factor (TGF)-β and tested for proliferation, activation, and expression of fibrosis-related factors. A dual luciferase reporter assay was performed for confirming the targeted relationship between miR-34b-5p and EZH2. Chromatin immunoprecipitation was used to measure EZH2 enrichment in the MFGE8 promoter region. We found that miR-34b-5p was lowly expressed in the CCl4-induced mouse model. Overexpression of miR-34b-5p suppressed both TGF-β-induced HSC proliferation and the expression of fibrosis-related factors and HSC activation markers. A dual luciferase assay showed a binding relationship between miR-34b-5p and EZH2. Overexpression of miR-34b-5p reduced TGF-β-induced HSC activation by inhibiting EZH2 to promote MFGE8 expression. Overexpression of miR-34b-5p inhibited liver fibrosis in vivo through the EZH2/MFGE8 axis. Conclusively, overexpressing miR-34b-5p reduced TGF-β-induced HSC activation by inhibiting EZH2 and thereby promoting MFGE8 expression, and inhibited liver fibrosis in vivo through the EZH2/MFGE8 axis.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We acknowledge and appreciate our colleagues for their valuable efforts and comments on this paper.

Funding

The grants were provided from the National Natural Science Foundation of Hunan Province (No.2022JJ30825), the National Natural Science Foundation of China (Nos. 81974079, 81500455, 32100597, 82070646), the National Key R&D Program of China (No. 2019YFE0190800) and the Key R&D Program of Hunan Province (No. 2020SK2083).

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Authors and Affiliations

  1. Department of Infectious Diseases, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, People’s Republic of China

    Jing Ma, Qiyao Liu, Mengxuan Chen, Bo He, Yi Li, Min Zhang & Yongfang Jiang

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  1. Jing Ma
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  2. Qiyao Liu
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  3. Mengxuan Chen
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Contributions

The authors declare that all the data were generated in-house and that no paper mill was used. MJ and JYF conceived the ideas. MJ and JYF designed the experiments. MJ, LQY, and CMX performed the experiments. MJ, JYF, HB, and LY analyzed the data. MJ, JYF, and ZM provided the critical materials. LQY and CMX wrote the manuscript. MJ and JYF supervised the study. All the authors have read and approved the final version for publication.

Corresponding author

Correspondence to Yongfang Jiang.

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Ethics approval and consent to participate

The animals were housed per the protocols permitted by the Institutional Animal Care and Use Committee of The Second Xiangya Hospital. All the experimental protocols obtained permission from the Animal Use Ethics Committee of The Second Xiangya Hospital (No. 2021524).

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All the authors agreed with the content and all gave explicit consent to submit and that they obtained consent from the responsible authorities at the institute/organization where they work, before the work is submitted.

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The authors declare no competing interests.

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Key points

• MiR-34b-5p is lowly expressed and EZH2 is highly expressed in liver fibrosis.

• Overexpression of miR-34b-5p inhibits hepatic stellate cell activation.

• MiR-34b-5p represses EZH2 expression.

• EZH2 represses MEGF8 expression through histone methylation.

• MiR-34b-5p/EZH2/MEGF8 affects hepatic stellate cell activation and liver fibrosis.

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Ma, J., Liu, Q., Chen, M. et al. MicroRNA-34b-5p binds enhancer of zeste 2 to inhibit milk fat globule-EGF factor 8 expression, affecting liver fibrosis. J Physiol Biochem 78, 885–895 (2022). https://doi.org/10.1007/s13105-022-00914-4

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  • DOI: https://doi.org/10.1007/s13105-022-00914-4

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