HJC0416 Attenuates Fibrogenesis in Activated Hepatic Stellate Cells via STAT3 and NF-κB Pathways.
Hepatic fibrosis
NF-κB
STAT3
Journal
The Journal of surgical research
ISSN: 1095-8673
Titre abrégé: J Surg Res
Pays: United States
ID NLM: 0376340
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
received:
28
02
2020
revised:
31
10
2020
accepted:
04
12
2020
pubmed:
25
1
2021
medline:
16
9
2021
entrez:
24
1
2021
Statut:
ppublish
Résumé
Hepatic fibrosis is wound-healing response that is the result of hepatic stellate cell (HSC) activation and subsequent excess extracellular matrix deposition. HSCs can be activated by a variety of inflammatory stimuli as well as through the signal transducer and activator of transcription 3 (STAT3) pathway. HJC0416 is a novel, orally bioavailable small-molecule inhibitor of STAT3 that was developed by our team using a fragment-based drug design approach. Previously, our team has shown that HJC0416 has antifibrogenic effects in activated HSCs. Recently, increasing evidence suggests that nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) plays an important role in the activation of HSCs. In the present study, we examined the role of NF-κB inhibition of HSC activation by HJC0416. LX-2 (human) and HSC-T6 (rat) cell lines were used. Expression levels of extracellular proteins, NF-κB and STAT3 expression and DNA binding, and inflammatory cytokine levels were determined using western blot, ELISA, and immunofluorescence assay. HJC0416 decreased cell viability in a dose-dependent manner in both cell lines and arrested the cell cycle at the S phase. Increased apoptosis was seen in LX-2 cells through Yo-Pro-1 and propidium iodide immunofluorescent stating. HJC0416 significantly decreased expression of fibronectin and collagen I as well as markedly decreased α-SMA and laminin. HJC0416 inhibited the STAT3 pathway by decreasing phosphorylation of STAT3, as well as signal transduction pathway activation. Notably, HJC0416 also inhibited the classic and alternative pathways of NF-κB activation. HJC0416 inhibited LPS-induced p65 nuclear translocation and DNA binding, as well as prevented phosphorylation and degradation of inhibitory protein IκBα. HJC0416 also prevented phosphorylation of serine residue 536 on p65. HJC0416, an inhibitor of STAT3, was found to have antifibrogenic properties in activated hepatic stellate cell lines. In addition, HJC0416 was found to inhibit the NF-κB pathway. Owing to this double effect, HJC0416 demonstrates promise for in vivo experimentation as an antifibrosis treatment.
Sections du résumé
BACKGROUND
Hepatic fibrosis is wound-healing response that is the result of hepatic stellate cell (HSC) activation and subsequent excess extracellular matrix deposition. HSCs can be activated by a variety of inflammatory stimuli as well as through the signal transducer and activator of transcription 3 (STAT3) pathway. HJC0416 is a novel, orally bioavailable small-molecule inhibitor of STAT3 that was developed by our team using a fragment-based drug design approach. Previously, our team has shown that HJC0416 has antifibrogenic effects in activated HSCs. Recently, increasing evidence suggests that nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) plays an important role in the activation of HSCs. In the present study, we examined the role of NF-κB inhibition of HSC activation by HJC0416.
METHODS
LX-2 (human) and HSC-T6 (rat) cell lines were used. Expression levels of extracellular proteins, NF-κB and STAT3 expression and DNA binding, and inflammatory cytokine levels were determined using western blot, ELISA, and immunofluorescence assay.
RESULTS
HJC0416 decreased cell viability in a dose-dependent manner in both cell lines and arrested the cell cycle at the S phase. Increased apoptosis was seen in LX-2 cells through Yo-Pro-1 and propidium iodide immunofluorescent stating. HJC0416 significantly decreased expression of fibronectin and collagen I as well as markedly decreased α-SMA and laminin. HJC0416 inhibited the STAT3 pathway by decreasing phosphorylation of STAT3, as well as signal transduction pathway activation. Notably, HJC0416 also inhibited the classic and alternative pathways of NF-κB activation. HJC0416 inhibited LPS-induced p65 nuclear translocation and DNA binding, as well as prevented phosphorylation and degradation of inhibitory protein IκBα. HJC0416 also prevented phosphorylation of serine residue 536 on p65.
CONCLUSIONS
HJC0416, an inhibitor of STAT3, was found to have antifibrogenic properties in activated hepatic stellate cell lines. In addition, HJC0416 was found to inhibit the NF-κB pathway. Owing to this double effect, HJC0416 demonstrates promise for in vivo experimentation as an antifibrosis treatment.
Identifiants
pubmed: 33486415
pii: S0022-4804(20)30906-9
doi: 10.1016/j.jss.2020.12.045
pmc: PMC8634552
mid: NIHMS1665697
pii:
doi:
Substances chimiques
(5-chloro-N-(1,1-dioxo-1H-1lamda(6)-benzo(b)thiophen-6-yl)-2-hydroxybenzamide
0
Benzamides
0
NF-kappa B
0
STAT3 Transcription Factor
0
Thiophenes
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
334-342Subventions
Organisme : NIGMS NIH HHS
ID : T32 GM008256
Pays : United States
Informations de copyright
Published by Elsevier Inc.
Références
Gut. 2018 Sep;67(9):1704-1715
pubmed: 28754776
Proc Natl Acad Sci U S A. 2015 Jun 9;112(23):7243-8
pubmed: 26039995
J Hepatol. 2011 Nov;55(5):1086-94
pubmed: 21354232
Eur J Pharmacol. 2015 Jan 15;747:71-87
pubmed: 25498792
FEBS J. 2015 Jul;282(14):2600-11
pubmed: 25825152
PLoS One. 2017 May 4;12(5):e0176173
pubmed: 28472150
J Clin Invest. 2013 May;123(5):1887-901
pubmed: 23635787
Int J Mol Sci. 2019 Mar 14;20(6):
pubmed: 30875826
Mol Cell. 2010 Aug 27;39(4):493-506
pubmed: 20797623
J Biol Chem. 2007 Nov 30;282(48):34958-67
pubmed: 17855361
Cell Prolif. 2020 Mar;53(3):e12777
pubmed: 32022328
Hepatology. 1997 Feb;25(2):361-7
pubmed: 9021948
Ital J Anat Embryol. 2004 Oct-Dec;109(4):225-38
pubmed: 15717457
Eur J Med Chem. 2013 Apr;62:498-507
pubmed: 23416191
Matrix Biol. 2018 Aug;68-69:452-462
pubmed: 29221811
Nat Rev Gastroenterol Hepatol. 2011 Feb;8(2):108-18
pubmed: 21293511
J Biol Chem. 2004 Dec 31;279(53):55633-43
pubmed: 15489227
Curr Opin Biotechnol. 2006 Dec;17(6):643-52
pubmed: 17084612
Cell. 2008 Aug 22;134(4):657-67
pubmed: 18724938
Biochem Pharmacol. 2004 Dec 15;68(12):2367-78
pubmed: 15548383
Antioxid Redox Signal. 2011 Jul 15;15(2):461-83
pubmed: 21194387
Trends Mol Med. 2008 Mar;14(3):109-19
pubmed: 18261959
Cancer Cell. 2009 Feb 3;15(2):79-80
pubmed: 19185839
Apoptosis. 2005 Oct;10(5):927-39
pubmed: 16151628
BMC Med. 2014 Sep 18;12:159
pubmed: 25286285
Cancer Manag Res. 2018 Dec 12;10:6857-6867
pubmed: 30588091
RSC Adv. 2016;6(102):100652-100663
pubmed: 28546859
Curr Opin Chem Biol. 2011 Aug;15(4):489-96
pubmed: 21665521
Hepatology. 2004 Feb;39(2):273-8
pubmed: 14767974
Hepatology. 2012 Sep;56(3):1150-9
pubmed: 22473749
Hepatology. 2016 Sep;64(3):955-65
pubmed: 26773297
J Surg Res. 2018 Dec;232:283-292
pubmed: 30463731
Hepatology. 2003 Mar;37(3):653-64
pubmed: 12601363
Semin Liver Dis. 2001 Aug;21(3):417-26
pubmed: 11586469
Eur J Med Chem. 2014 Jul 23;82:195-203
pubmed: 24904966
Best Pract Res Clin Gastroenterol. 2011 Apr;25(2):195-206
pubmed: 21497738
Hepatology. 2009 Feb;49(2):533-44
pubmed: 19115220
PLoS One. 2017 Mar 30;12(3):e0174374
pubmed: 28358817
Int J Mol Sci. 2018 May 24;19(6):
pubmed: 29795016
J Immunol. 2015 Apr 1;194(7):3422-31
pubmed: 25740948
J Hepatol. 2009 Jul;51(1):139-48
pubmed: 19457567
Histol Histopathol. 2010 Aug;25(8):1075-91
pubmed: 20552556
Nature. 2011 Aug 03;476(7358):96-100
pubmed: 21814282
J Biol Chem. 2013 Jan 4;288(1):285-93
pubmed: 23100252
J Hepatol. 2011 Sep;55(3):612-625
pubmed: 21251937
Int J Oncol. 2014 Apr;44(4):1032-40
pubmed: 24430672
Biochim Biophys Acta. 2013 Jul;1832(7):948-54
pubmed: 23470555
Hepatology. 1996 Sep;24(3):670-6
pubmed: 8781341