4-methylumbelliferone Prevents Liver Fibrosis by Affecting Hyaluronan Deposition, FSTL1 Expression and Cell Localization.

Irina N Andreichenko,Elena V Bulgakova,Alexandra A Tsitrina,Alexey M Kulikov,Arsen S Mikaelyan,Dmitry I Maltsev,Yuri V Kotelevtsev,Alexander V Fokin,Grigorii S Perelman,Adelya I Gabdulkhakova

doi:10.3390/ijms20246301

Abstract

4-methylumbelliferone (4MU) is an inhibitor of hyaluronan deposition and an active substance of hymecromone, a choleretic and antispasmodic drug. 4MU reported to be anti-fibrotic in mouse models; however, precise mechanism of action still requires further investigation. Here we describe the cellular and molecular mechanisms of 4MU action on CCl4-induced liver fibrosis in mice using NGS transcriptome, Q-PCR and immunohistochemical analysis. Collagen and hyaluronan deposition were prevented by 4MU. The CCl4 stimulated expression of Col1a and αSMA were reduced, while the expression of the ECM catabolic gene Hyal1 was increased in the presence of 4MU. Bioinformatic analysis identified an activation of TGF-beta and Wnt/beta-catenin signaling pathways, and inhibition of the genes associated with lipid metabolism by CCL4 treatment, while 4MU restored key markers of these pathways to the control level. Immunohistochemical analysis reveals the suppression of hepatic stellate cells (HSCs) transdifferentiation to myofibroblasts by 4MU treatment. The drug affected the localization of HSCs and macrophages in the sites of fibrogenesis. CCl4 treatment induced the expression of FSTL1, which was downregulated by 4MU. Our results support the hypothesis that 4MU alleviates CCl4-induced liver fibrosis by reducing hyaluronan deposition and downregulating FSTL1 expression, accompanied by the suppression of HSC trans-differentiation and altered macrophage localization.

Highlights

Extracellular matrix (ECM) deposition is a natural reparative response to the liver injury caused by chemical toxins or viral infections
For detail characterization of 4MU effect on fibrogenesis, we evaluated the Tresholded Manders’ coefficients for the colocalization of HAS2 on macrophage (F4/80) on hepatic stellate cells (HSCs) (GFAP)
We report the first time that small molecule, 4MU, which downregulates mRNA level and protein expression of follistatin-like protein 1 (FSTL1) in the settings of family [24]

Summary

Introduction

Extracellular matrix (ECM) deposition is a natural reparative response to the liver injury caused by chemical toxins or viral infections. Fibrosis occurs as an excessive formation of ECM resulting in the subsequent growth of connective tissue in the affected areas. Collagen 1 and 3 are the major components of the fibrous formations in the liver. They are produced by myofibroblasts, which originate predominantly from the hepatic stellate cells (HSCs), pericyte like cells expressing desmin and glial fibrillary acidic protein (GFAP). In the quiescent state HSCs are uniformly spread throughout the liver parenchyma, providing storage for retinol.

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