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. 2019 Dec 20;9(1):30.
doi: 10.3390/cells9010030.

(-)-Catechin-7- O-β-d-Apiofuranoside Inhibits Hepatic Stellate Cell Activation by Suppressing the STAT3 Signaling Pathway

Yong Joo Park  1 , Dong Min Kim  1 , Mi Ho Jeong  1 , Jae Sik Yu  1 , Hae Min So  1 , In Jae Bang  1 , Ha Ryong Kim  2 , Seung-Hwan Kwon  3   4 , Ki Hyun Kim  1 , Kyu Hyuck Chung  1
Affiliations

(-)-Catechin-7- O-β-d-Apiofuranoside Inhibits Hepatic Stellate Cell Activation by Suppressing the STAT3 Signaling Pathway

Yong Joo Park et al. Cells. .

Abstract

Hepatic fibrosis is characterized by the abnormal deposition of extracellular matrix (ECM) proteins. During hepatic fibrogenesis, hepatic stellate cell (HSC) activation followed by chronic injuries is considered a key event in fibrogenesis, and activated HSCs are known to comprise approximately 90% of ECM-producing myofibroblasts. Here, we demonstrated that (-)-catechin-7-O-β-d-apiofuranoside (C7A) significantly inhibited HSC activation via blocking the signal transducer and activator of transcription 3 (STAT3) signaling pathway. This is the first study to show the hepatic protective effects of C7A with possible mechanisms in vitro and in vivo. In our bioactivity screening, we figured out that the EtOH extract of Ulmusdavidiana var. japonica root barks, which have been used as a Korean traditional medicine, inhibited collagen synthesis in HSCs. Four catechins isolated from the EtOAc fraction of the EtOH extract were compared with each other in terms of reduction in collagen, which is considered as a marker of hepatic protective effects, and C7A showed the strongest inhibitory effects on HSC activation in protein and qPCR analyses. As a possible mechanism, we investigated the effects of C7A on the STAT3 signaling pathway, which is known to activate HSCs. We found that C7A inhibited phosphorylation of STAT3 and translocation of STAT3 to nucleus. C7A also inhibited expressions of MMP-2 and MMP-9, which are downstream genes of STAT3 signaling. Anti-fibrotic effects of C7A were evaluated in a thioacetamide (TAA)-induced liver fibrosis model, which indicated that C7A significantly inhibited ECM deposition through inhibiting STAT3 signaling. C7A decreased serum levels of aspartate amino transferase and alanine transaminase, which were markedly increased by TAA injection. Moreover, ECM-associated proteins and mRNA expression were strongly suppressed by C7A. Our study provides the experimental evidence that C7A has inhibitory effects on HSC activation after live injury and has preventive and therapeutic potentials for the management of hepatic fibrosis.

Keywords: (–)-Catechin-7-O-β-d-apiofuranoside; STAT3; Ulmus davidiana var. japonica; hepatic fibrosis; hepatic stellate cells.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
EtOH extract and EtOAc fraction of Ulmus davidiana var. japonica suppress the fibrotic effect in activated LX-2 cells. LX-2 cells were treated with EtOH extract or EtOAc fraction of U. davidiana var. japonica for 48 h after TGF-β1 induction for 48 h. Cytotoxicities of (A) EtOH extract and (C) EtOAc fraction in LX-2 cells were investigated by WST-1 assay after treatment for 48 h. The protein expression of collagen was analyzed by western blot assay in (B) EtOH extract and (D) EtOAc fraction treated groups. GAPDH was used as a loading control. Each experiment was repeated three times, and values represent mean ± S.D. * p < 0.05 compared with control.
Figure 2
Figure 2
Chemical structures of catechins (1–4) isolated from U. davidiana var. japonica.
Figure 3
Figure 3
Antifibrotic effects of compounds (1–4) from EtOAc fraction in activated LX-2 cells. The inhibitory effects of compounds (1–4) were tested by treating them for 48 h, respectively, after TGF-β1 induction for 48 h. (A) Cytotoxicity of compounds (1–4) in LX-2 cells was evaluated by WST-1 assay after treatment for 48 h. (B) The protein expressions of collagen were analyzed by western blot assay in each compound treated group. (C) Relative collagen mRNA expressions were analyzed by qPCR analysis. Each experiment was repeated three times, and values represent mean ± S.D. ## p < 0.01 compared with control, ** p < 0.01, * p < 0.05 compared with TGF-β1 treatment group.
Figure 4
Figure 4
(–)-Catechin-7-O-β-d-apiofuranoside (C7A) suppresses the activation of LX-2 cells. Inhibitory effects of C7A on LX-2 cell activation were tested by treating it for 48 h after TGF-β1 induction for 48 h. (A) The protein expressions of fibronectin, α-SMA, and CTGF were analyzed by western blot assay, and relative protein expressions were obtained from Image J quantification values. (B) The mRNA expression levels of fibronectin, α-SMA, and CTGF were evaluated by qPCR analysis. Each experiment was repeated three times, and values represent mean ± S.D. ## p < 0.01 compared with control, ** p < 0.01, * p < 0.05 compared with TGF-β1 treatment group.
Figure 5
Figure 5
C7A suppresses STAT3 phosphorylation and translocation in TGF-β1-activated LX-2 cells. LX-2 cells were treated with C7A for 48 h after TGF-β1 induction for 48 h. (A) The expression levels of p-STAT3 and STAT3 were analyzed by western blot assay. (B) The expression levels of p-STAT3 were analyzed from nuclear and cytosolic protein fractions. Lamin A/C was used as a nuclear loading control. (C) p-STAT3 (green) localization in LX-2 cells was determined by confocal immunocytochemistry. The nuclei are counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (Blue). The scale bars represent 50 µm. (D) The protein expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured by western blot assay. Densitometric analysis is expressed as mean ± SD intensity of optical density obtained by three independent experiments. (E) The mRNA expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were evaluated by qPCR analysis. Each experiment was repeated three times, and values represent mean ± S.D. ## p < 0.01 compared with control, ** p < 0.01, * p < 0.05 compared with TGF-β1 treatment group.
Figure 5
Figure 5
C7A suppresses STAT3 phosphorylation and translocation in TGF-β1-activated LX-2 cells. LX-2 cells were treated with C7A for 48 h after TGF-β1 induction for 48 h. (A) The expression levels of p-STAT3 and STAT3 were analyzed by western blot assay. (B) The expression levels of p-STAT3 were analyzed from nuclear and cytosolic protein fractions. Lamin A/C was used as a nuclear loading control. (C) p-STAT3 (green) localization in LX-2 cells was determined by confocal immunocytochemistry. The nuclei are counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (Blue). The scale bars represent 50 µm. (D) The protein expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured by western blot assay. Densitometric analysis is expressed as mean ± SD intensity of optical density obtained by three independent experiments. (E) The mRNA expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were evaluated by qPCR analysis. Each experiment was repeated three times, and values represent mean ± S.D. ## p < 0.01 compared with control, ** p < 0.01, * p < 0.05 compared with TGF-β1 treatment group.
Figure 6
Figure 6
C7A attenuated thioacetamide (TAA)-induced chronic liver fibrosis. (A) Mice were given intraperitoneal injections for three weeks of saline or C7A (40 mg/kg) after 3 weeks of saline or TAA (150 mg/kg) treatment. (B) Serum aspartate aminotransferase (AST) and alanine transaminase (ALT)levels were analyzed. (C) Representative histology section of liver tissues stained with H&E and Masson’s trichrome. (black arrow: collagen deposition; scale bar = 500 μm). (D) Liver hydroxyproline contents per gram of liver tissue of mice from each group. (E) Gene expressions of α-SMA, Col1A1, Col3A1, and CTGF were measured by qPCR analysis. (F) Western blotting of fibronectin, α-SMA, p-STAT3, and GAPDH in the livers of mice from each group. Each experiment was repeated three times, and values represent mean ± S.D. (n = 7) ## p < 0.01 compared with control, * p < 0.05, ** p < 0.01 compared with TAA treatment group.
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References

    1. Hui A.Y., Friedman S.L. Molecular basis of hepatic fibrosis. Expert Rev. Mol. Med. 2003;5:1–23. doi: 10.1017/S1462399403005684. - DOI - PubMed
    1. Geerts A. History, heterogeneity, developmental biology, and functions of quiescent hepatic stellate cells. Semin. Liver Dis. 2001;21:311–335. doi: 10.1055/s-2001-17550. - DOI - PubMed
    1. Bataller R., Brenner D.A. Liver fibrosis. J. Clin. Investig. 2005;115:209–218. doi: 10.1172/JCI24282. - DOI - PMC - PubMed
    1. Friedman S.L. Mechanisms of hepatic fibrogenesis. Gastroenterology. 2008;134:1655–1669. doi: 10.1053/j.gastro.2008.03.003. - DOI - PMC - PubMed
    1. Friedman S.L. Hepatic stellate cells: Protean, multifunctional, and enigmatic cells of the liver. Physiol. Rev. 2008;88:125–172. doi: 10.1152/physrev.00013.2007. - DOI - PMC - PubMed

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