Sains Malaysiana 54(3)(2025): 881-898

http://doi.org/10.17576/jsm-2025-5403-20

let-7g-5p Suppresses the Proliferation and Expansion of Hepatic Stellate Cells in Liver Fibrosis via Targeting FGF5

(let-7g-5p Mencegah Pembiakan dan Pengembangan Sel Stellate Hepatik dalam Fibrosis Hati melalui Penyasaran FGF5)

JIAMING ZHOU^1,2,3†,*, SIYAN HUANG^1,2†, YIDAN YANG^1,2, YUTONG CONG², YUWEN CHEN², YOUYOU DING², MENGWEN LU² & MEIYUAN LI^1,2

¹Department of Pathology, Medical School of Nantong University, Nantong 226001, Jiangsu, China

²Department of Medicine, Xinglin College of Nantong University, Nantong 226001, Jiangsu, China

³Department of Pathology, Second Affiliated Hospital of Naval Medical University, Shanghai 200003, China

Diserahkan: 28 April 2024/Diterima: 13 Disember 2024

†Jiaming Zhou and Siyan Huang contributed equally to this work

Abstract

Hepatic stellate cells (HSCs) and their activated phenotype (activated HSCs, aHSCs) function as crucial effector cells in the onset of liver fibrosis. In recent years, microRNAs (miRNAs) have emerged as a promising therapeutic approach for diseases. To explore early intervention strategies for HSCs activation and proliferation, miRNA profiles were sequenced from three patients with chronic hepatitis B-related hepatic fibrosis (HF). The miRNAs sequencing experiment and data analysis were conducted utilizing the Illumina HiSeq 4000 platform. In vitro, the proliferation and expansion capabilities of HSCs were detected using CCK8, EdU and colony formation assays. The examination of α-SMA, the indicator aHSCs, was performed through western blot assays. For in vivo investigation of the let-7g-5p/FGF5 axis, a bile duct ligation (BDL)-induced HF mice model was constructed and mmu-let-7g-5p agomir was delivered to mice via tail vein injection. Collagen deposition and the α-SMA level were assessed through histological staining including H&E, Masson, Van Gieson(VG), and immunohistochemical (IHC) staining.The miRNAs sequencing and bioinformatics analysis identified let-7g-5p as a promising anti-HF candidate. qRT-PCR and dual-luciferase reporter assays confirmed FGF5 as the direct target of let-7g-5p. let-7g-5p hampered the proliferation and expansion abilities of HSCs and decreased the α-SMA level by targeting FGF5 in vitro. In addition, H&E, Masson, VG and IHC staining demonstrated the let-7g-5p/FGF5 axis significantly mitigated collagen deposition and decreased α-SMA production in the BDL-induced HF mice. let-7g-5p suppressed the proliferation and expansion of HSCs and alleviated HF via targeting FGF5. The let-7g-5p/FGF5 axis could be an effective therapeutic target for reducing aHSCs abundance in HF.

Keywords: Bile duct ligation; FGF5; hepatic fibrosis; hepatic stellate cells; let-7g-5p; proliferation; α-SMA

Abstrak

Sel bintang hepatik (HSC) dan fenotip yang diaktifkan (HSC diaktifkan, aHSC) berfungsi sebagai sel efektor penting dalam permulaan fibrosis hati. Dalam beberapa tahun kebelakangan ini, mikroRNA (miRNA) telah muncul sebagai pendekatan terapeutik yang berpotensi untuk penyakit. Untuk meneroka strategi intervensi awal untuk pengaktifan dan percambahan HSC, profil miRNA telah dijujukan daripada tiga pesakit dengan fibrosis hepatik (HF) berkaitan hepatitis B kronik. Uji kaji penjujukan miRNA dan analisis data telah dijalankan menggunakan platform Illumina HiSeq 4000. Secara in vitro, keupayaan percambahan dan pengembangan HSC telah dikesan menggunakan CCK8, EdU dan ujian pembentukan koloni. Pemeriksaanα-SMA, penunjuk aHSC, telah dilakukan melalui ujian pemblotan western. Untuk kajian in vivo paksi let-7g-5p/FGF5, model tikus HF yang disebabkan oleh pengikatan saluran hempedu (BDL) telah dibina dan mmu-let-7g-5p agomir dihantar kepada tikus melalui suntikan urat ekor. Pemendapan kolagen dan tahapα-SMA dinilai melalui pewarnaan histologi termasuk pewarnaan H&E, Masson, Van Gieson(VG) dan pewarnaan imunohistokimia (IHC). Penjujukan miRNA dan analisis bioinformatik mengenal pasti let-7g-5p sebagai calon anti-HF yang berpotensi. Ujian reporter qRT-PCR dan dwi-luciferase mengesahkan FGF5 sebagai sasaran langsung let-7g-5p. let-7g-5p menghalang kebolehan percambahan dan pengembangan HSC dan mengurangkan tahapα-SMA dengan menyasarkan FGF5 secara in vitro. Di samping itu, pewarnaan H&E, Masson, VG dan IHC menunjukkan paksi let-7g-5p/FGF5 dengan ketara mengurangkan pemendapan kolagen dan mengurangkan pengeluaranα-SMA pada tikus HF yang disebabkan oleh BDL. let-7g-5p menyekat percambahan dan pengembangan HSC dan mengurangkan HF melalui menyasarkan FGF5. Paksi let-7g-5p/FGF5 boleh menjadi sasaran terapeutik yang berkesan untuk mengurangkan kelimpahan aHSC dalam HF.

Kata kunci: FGF5; fibrosis hepatik; let-7g-5p; ligasi saluran hempedu; percambahan; sel bintang hepatik; α-SMA

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*Pengarang untuk surat-menyurat; email: zhoujiaming@ntu.edu.cn