microRNA-322/424 promotes liver fibrosis by regulating angiogenesis through targeting CUL2/HIF-1α pathway

Qinglan Wang; Feng Zhang; Yang Lei; Ping Liu; Chenghai Liu; Yanyan Tao

doi:10.1016/j.lfs.2020.118819

microRNA-322/424 promotes liver fibrosis by regulating angiogenesis through targeting CUL2/HIF-1α pathway

Life Sci. 2021 Feb 1:266:118819. doi: 10.1016/j.lfs.2020.118819. Epub 2020 Dec 14.

Authors

Qinglan Wang¹, Feng Zhang², Yang Lei², Ping Liu³, Chenghai Liu⁴, Yanyan Tao⁵

Affiliations

¹ Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China; College of Basic Medical Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; E-Institute of Shanghai Municipal Education Committee, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
² College of Basic Medical Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
³ Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China; E-Institute of Shanghai Municipal Education Committee, Shanghai University of Traditional Chinese Medicine, Shanghai, China. Electronic address: liuliver@vip.sina.com.
⁴ Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China; E-Institute of Shanghai Municipal Education Committee, Shanghai University of Traditional Chinese Medicine, Shanghai, China; Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai, China; Shanghai Innovation Center of Traditional Chinese Medicine Health Service, Shanghai, China. Electronic address: chenghailiu@hotmail.com.
⁵ Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China; Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai, China. Electronic address: taoyanyan1023@126.com.

PMID: 33333053
DOI: 10.1016/j.lfs.2020.118819

Abstract

Aims: To investigate the effects and mechanism of miR-322/424 in liver fibrosis.

Main methods: miR-322/424 expression in liver cirrhosis patients, mouse and rat liver fibrosis was determined by qPCR. Mice liver fibrosis was established by CCl₄, and intervened by miR-322/424 agomir or antagomir. Liver hydroxyproline content and Sirius red staining were used to evaluate collagen deposition. CD31 expression was used to evaluate liver microvessel density. In vitro, the effects of miR-322/424 mimic or inhibitor on human hepatic sinusoidal endothelial cells (HHSECs) migration and tube formation were investigated. A dual luciferase reporter assay was performed to confirm the direct interaction between miR-322/424 and Cullin2. mRNA expression of elongin B/C, Cullin2, and RBX1 was determined by qPCR. HIF-1α protein expression was determined by Western blotting.

Key findings: miR-322/424 level in liver cirrhosis patients, mouse liver fibrosis induced by CCl₄ and BDL, and rat liver fibrosis induced by CCl₄ and dimethylnitrosamine was increased. miR-322/424 agomir exacerbated CCl₄-induced mouse liver fibrosis, whereas the opposite effect was observed for miR-322/424 antagomir. miR-322/424 agomir significantly upregulated liver CD31 expression; opposite effects occurred with miR-322/424 antagomir. In vitro, miR-322/424 mimic significantly promoted tube formation and cell migration, and increased von Willebrand factor expression, whereas miR-322/424 inhibitor had the opposite effect. Dual-Luciferase Reporter Assay identified Cullin2 as miR-322/424 target. miR-322/424 decreased the mRNA expression of elongin B/C, Cullin2, and RBX1 and increased HIF-1α protein expression in HHSECs.

Significance: miR-322/424 plays a central role in the pathogenesis of liver fibrosis by targeting Cullin2, and enhancing HIF-1α-mediated hepatic angiogenesis.

Keywords: Hepatic angiogenesis; Liver fibrosis; miR-322/424; microRNA.

MeSH terms

Animals
Cells, Cultured
Cullin Proteins / genetics
Cullin Proteins / metabolism*
Disease Models, Animal
Hemangioma / genetics
Hemangioma / metabolism
Hemangioma / pathology*
Humans
Hypoxia-Inducible Factor 1, alpha Subunit / genetics
Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
Liver Cirrhosis / genetics
Liver Cirrhosis / metabolism
Liver Cirrhosis / pathology*
Male
Mice
Mice, Inbred C57BL
Mice, Inbred ICR
MicroRNAs / genetics*
Neovascularization, Pathologic / genetics
Neovascularization, Pathologic / metabolism
Neovascularization, Pathologic / pathology*
Rats
Rats, Wistar

Substances

CUL2 protein, human
Cullin Proteins
HIF1A protein, human
Hypoxia-Inducible Factor 1, alpha Subunit
MIRN424 microrna, human
MicroRNAs