miR‑146b‑5p activation of hepatic stellate cells contributes to the progression of fibrosis by directly targeting HIPK1

Xie,Junfeng; Cheng,Na; Huang,Zhanchao; Shu,Xu; Xiang,Tianxin

doi:10.3892/etm.2022.11474

miR‑146b‑5p activation of hepatic stellate cells contributes to the progression of fibrosis by directly targeting HIPK1

Authors:
- Junfeng Xie
- Na Cheng
- Zhanchao Huang
- Xu Shu
- Tianxin Xiang
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Affiliations: Department of Hospital Infection Control, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330000, P.R. China, Department of Infectious Disease, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330000, P.R. China
Published online on: June 27, 2022 https://doi.org/10.3892/etm.2022.11474
Article Number: 537
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to explore the biological functions of microRNA (miR)‑146b‑5p and homeodomain interacting protein kinase 1 (HIPK1) in the progression of hepatic fibrosis (HF) and to identify the underlying mechanism. A rat HF model was established by administering a subcutaneous injection of carbon tetrachloride (CCl₄). Relative levels of miR‑146b‑5p and HIPK1 in fibrotic rat liver tissues and the rat hepatic stellate cell (HSC) line HSC‑T6 were measured by quantitative reverse transcription PCR, western blotting and immunohistochemistry. Following activation of HSC‑T6 cells by lipopolysaccharide (LPS) induction, cell viability was examined by MTT assay. Transfection of miR‑146b‑5p mimic or inhibitor into HSC‑T6 cells was performed, with the aim to identify the influence of miR‑146b‑5p on HSC‑T6 cell behavior. The targeting relationship between miR‑146b‑5p and HIPK1 was predicted by TargetScan 7.2 and StarBase 3.0 and it was later verified by a dual‑luciferase reporter assay. Through lentivirus transfection, the biological function of HIPK1 in regulating the progression of HF and the underlying mechanism were investigated. The results showed that miR‑146b‑5p was upregulated in liver tissues of rats with HF and activated HSC‑T6 cells, while HIPK1 was downregulated in liver tissues of rats with HF and activated HSC‑T6 cells. miR‑146b‑5p was able to upregulate the activation markers of LPS‑induced HSC‑T6 cells, upregulate COL1A1 and TGF‑β, increase cell viability and contribute to fibrosis progression. HIPK1 was validated as the direct target of miR‑146b‑5p and its overexpression could effectively reduce the effect of miR‑146b‑5p in contribution to the progression of HF. In conclusion, miR‑146b‑5p was significantly upregulated during the progression of HF. By targeting and downregulating HIPK1, miR‑146b‑5p could significantly activate HSCs, upregulate COL1A1 and TGF‑β and contribute to fibrosis progression. miR‑146b‑5p is a potential biomarker and therapeutic target for HF.

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