M1567 CpG Motifs Induce Proinflammatory Events in Hepatic Stellate Cells and Are Involved in Bile Duct Ligation-Induced Hepatic Fibrosis In Vivo

Exemestane (EXE) is an irreversible steroidal aromatase inhibitor mainly used as an adjuvant endocrine therapy for postmenopausal women suffering from breast cancer. Besides inhibiting aromatase activity, EXE has multiple biological functions, such as antiproliferation, anti-inflammatory, and antioxidant activities which are all involved in hepatic fibrosis. Therefore, we investigated the role of EXE during the progress of hepatic fibrosis. The effect of EXE on liver injury and fibrosis were assessed in two hepatic fibrosis rat models, which were induced by either carbon tetrachloride (CCl4) or bile duct ligation (BDL). The influence of EXE treatment on activation and proliferation of primary rat hepatic stellate cells (HSCs) was observedin vitro. The results showed that EXE attenuated the liver fibrosis by decreasing the collagen deposition andα-SMA expressionin vivoand inhibited the activation and proliferation of primary rat HSCsin vitro. Additionally, EXE promoted the secretion of antifibrotic and anti-inflammatory cytokine IL-10in vivoand in HSC-T6 culture media. In conclusion, our findings reveal a new function of EXE on hepatic fibrosis and prompted its latent application in liver fibrotic-related disease.

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iRhom2 inhibits bile duct obstruction–induced liver fibrosis

Science Signaling ◽

10.1126/scisignal.aax1194 ◽

2019 ◽

Vol 12 (605) ◽

pp. eaax1194 ◽

Cited By ~ 7

Author(s):

Balamurugan Sundaram ◽

Kristina Behnke ◽

Andrea Belancic ◽

Mazin A. Al-Salihi ◽

Yasser Thabet ◽

...

Keyword(s):

Bile Duct ◽

Liver Fibrosis ◽

Hepatic Stellate Cells ◽

Stellate Cells ◽

Duct Ligation ◽

Tnf Α ◽

Primary Mouse ◽

Factor Α

Chronic liver disease can induce prolonged activation of hepatic stellate cells, which may result in liver fibrosis. Inactive rhomboid protein 2 (iRhom2) is required for the maturation of A disintegrin and metalloprotease 17 (ADAM17, also called TACE), which is responsible for the cleavage of membrane-bound tumor necrosis factor–α (TNF-α) and its receptors (TNFRs). Here, using the murine bile duct ligation (BDL) model, we showed that the abundance of iRhom2 and activation of ADAM17 increased during liver fibrosis. Consistent with this, concentrations of ADAM17 substrates were increased in plasma samples from mice after BDL and in patients suffering from liver cirrhosis. We observed increased liver fibrosis, accelerated disease progression, and an increase in activated stellate cells after BDL in mice lacking iRhom2 (Rhbdf2−/−) compared to that in controls. In vitro primary mouse hepatic stellate cells exhibited iRhom2-dependent shedding of the ADAM17 substrates TNFR1 and TNFR2. In vivo TNFR shedding after BDL also depended on iRhom2. Treatment of Rhbdf2−/− mice with the TNF-α inhibitor etanercept reduced the presence of activated stellate cells and alleviated liver fibrosis after BDL. Together, these data suggest that iRhom2-mediated inhibition of TNFR signaling protects against liver fibrosis.

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Hepatic stellate cells retain retinoid-laden lipid droplets after cellular transdifferentiation into activated myofibroblasts

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00251.2017 ◽

2018 ◽

Vol 315 (5) ◽

pp. G713-G721 ◽

Cited By ~ 9

Author(s):

Loretta L. Jophlin ◽

Yiannis Koutalos ◽

Chunhe Chen ◽

Vijay Shah ◽

Don C. Rockey

Keyword(s):

Hepatic Stellate Cells ◽

Lipid Droplets ◽

Bile Duct Ligation ◽

Ex Vivo ◽

Stellate Cells ◽

Duct Ligation ◽

Oil Red O

Loss of retinyl ester (RE)-rich lipid droplets (LDs) from hepatic stellate cells (HSCs) is cited as a key event in their cellular transdifferentiation to activated, pro-fibrotic myofibroblasts; however, it remains unclear if changes in LD morphology or RE content are causal for transdifferentiation. To better understand LD dynamics in vitro within a common model of HSC activation, we used novel approaches preserving LD morphology and allowing for quantitation of RE. The size and quantity of LDs within in vitro and in vivo bile duct ligation (BDL)-activated HSCs were quantitated using adipocyte differentiation-related protein (ADRP) labeling and oil red o (ORO) staining (gold standard), and RE content was determined using fluorescence microscopy. We found during HSC activation in vitro that LD number differed significantly when measured by ADRP and ORO, respectively ( day 1: 56 vs. 5, P = 0.03; day 4: 101 vs. 39, P = 0.03; day 14: 241 vs. 12, P = 0.02). Ex vivo HSCs activated in vivo contained the same number of LDs as day 4 in vitro activated HSCs (118 vs. 101, P = 0.54). Decline in LD RE occurred beyond day 4 in vitro and day 1 ex vivo , after HSC transdifferentiation was underway. Lastly, in situ HSCs examined using electron microscopy show LDs tend to be smaller but are ultimately retained in BDL injured livers. Therefore, we conclude that during HSC transdifferentiation, LDs are not lost but are retained, decreasing in size. Additionally, RE content declines after transdifferentiation is underway. These data suggest that these LD changes are not causal for HSC transdifferentiation. NEW & NOTEWORTHY Loss of retinoid-laden lipid droplets from hepatic stellate cells has long been held as a hallmark of their transdifferentiation into activated myofibroblasts, the dominant cells that drive hepatic fibrosis. This study demonstrates that stellate cells activated in culture and after liver injury in vivo retain their lipid droplets and that these droplets become smaller and more numerous, with decreases in droplet retinoid concentration occurring only after cellular transdifferentiation is underway.

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3,5-Diethoxy-3′-Hydroxyresveratrol (DEHR) Ameliorates Liver Fibrosis via Caveolin-1 Activation in Hepatic Stellate Cells and in a Mouse Model of Bile Duct Ligation Injury

Molecules ◽

10.3390/molecules23112833 ◽

2018 ◽

Vol 23 (11) ◽

pp. 2833 ◽

Cited By ~ 3

Author(s):

Phil Lee ◽

Hye-Jin Park ◽

Namki Cho ◽

Hong Kim

Keyword(s):

Bile Duct ◽

Liver Fibrosis ◽

Western Blot ◽

Hepatic Stellate Cells ◽

Bile Duct Ligation ◽

B Cell Lymphoma ◽

Stellate Cells ◽

Transfected Cells ◽

Caveolin 1 ◽

Duct Ligation

Hepatic stellate cells (HSCs) are involved in the pathogenesis of liver fibrosis. Resveratrol, 3,5,4′-trihydroxystilbene, is a dietary polyphenol found in natural food products. Here, we evaluated the anti-proliferative effects of a synthetic resveratrol derivative, 3,5-diethoxy-3′-hydroxyresveratrol (DEHR), on HSCs. Flow cytometry and Western blot analyses showed that DEHR induces apoptosis through the upregulation of cleaved caspase-3 and poly (ADP-ribose) polymerase expression and reduction in the level of an anti-apoptotic protein B-cell lymphoma 2 (Bcl2). As caveolin-1 (CAV1), a competitive inhibitor of heme oxygenase 1 (HO-1), is related to apoptotic proteins in hepatic cells, we focused on the role of CAV1 in DEHR-induced apoptosis in HSCs through Western blot analyses. Our results showed that the inhibitory effect of DEHR on cell viability was stronger in HO-1 siRNA-transfected cells but weakened in CAV1 siRNA-transfected cells. Collagen concentration was significantly reduced, whereas CAV1 expression increased after treatment of a bile duct ligation injury-induced liver fibrosis model with DEHR for four weeks. We confirmed that DEHR treatment significantly reduced fibrous hyperplasia around the central veins, using hematoxylin and eosin and Sirius red staining. DEHR ameliorates liver fibrosis in vitro and in vivo, possibly through a mechanism involving CAV1.

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MicroRNA-29a Alleviates Bile Duct Ligation Exacerbation of Hepatic Fibrosis in Mice through Epigenetic Control of Methyltransferases

International Journal of Molecular Sciences ◽

10.3390/ijms18010192 ◽

2017 ◽

Vol 18 (1) ◽

pp. 192 ◽

Cited By ~ 24

Author(s):

Ya-Ling Yang ◽

Feng-Sheng Wang ◽

Sung-Chou Li ◽

Mao-Meng Tiao ◽

Ying-Hsien Huang

Keyword(s):

Bile Duct ◽

Hepatic Fibrosis ◽

Bile Duct Ligation ◽

Epigenetic Control ◽

Duct Ligation

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Preventive effects of ME3738 on hepatic fibrosis induced by bile duct ligation in rats

Hepatology Research ◽

10.1111/j.1872-034x.2008.00326.x ◽

2008 ◽

Vol 38 (7) ◽

pp. 727-735 ◽

Cited By ~ 10

Author(s):

Kazunori Maeda ◽

Masahiko Koda ◽

Tomomitsu Matono ◽

Takaaki Sugihara ◽

Satoru Yamamoto ◽

...

Keyword(s):

Bile Duct ◽

Hepatic Fibrosis ◽

Bile Duct Ligation ◽

Duct Ligation ◽

Preventive Effects

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Vitamin D inhibits development of liver fibrosis in an animal model but cannot ameliorate established cirrhosis

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00132.2013 ◽

2015 ◽

Vol 308 (2) ◽

pp. G112-G120 ◽

Cited By ~ 37

Author(s):

Shirley Abramovitch ◽

Efrat Sharvit ◽

Yosef Weisman ◽

Amir Bentov ◽

Eli Brazowski ◽

...

Keyword(s):

Vitamin D ◽

Growth Factor ◽

Bile Duct ◽

Liver Fibrosis ◽

Bile Duct Ligation ◽

Active Form ◽

Preventive Treatment ◽

Antifibrotic Effect ◽

Duct Ligation

1,25(OH)2D3, the active form of vitamin D, has an antiproliferative and antifibrotic effect on hepatic stellate cells. Our aim was to investigate the potential of 1,25(OH)2D3 to inhibit the development of liver fibrosis and to ameliorate established fibrosis in vivo. The antifibrotic effect of 1,25(OH)2D3 was investigated in a thioacetamide (TAA) model (as a preventive treatment and as a remedial treatment) and in a bile duct ligation model. In the preventive model, rats received simultaneously intraperitoneum injection of TAA and/or 1,25(OH)2D3 for 10 wk. In the remedial model, rats were treated with TAA for 10 wk and then received 1,25(OH)2D3 or saline for 8 wk. Fibrotic score was determined by Masson staining. Collagen I, α-smooth muscle actin (α-SMA), tissue inhibitor of metalloproteinase-1 (TIMP1), platelet-derived growth factor (PDGF), and transforming growth factor-β (TGF-β) expression were measured by Western blot analysis and real-time PCR. Hypercalemia was detected by chemistry measurements. Preventive treatment of 1,25(OH)2D3 significantly suppressed liver fibrosis both macroscopically and microscopically and significantly lowered the fibrotic score of the TAA + 1,25(OH)2D3 group compared with the TAA group. 1,25(OH)2D3 significantly inhibited expression of PDGF and TGF-β by ∼50% and suppressed the expression of collagen Iα1, TIMP1, and α-SMA by approximately three-, two-, and threefold, respectively. In contrast, 1,25(OH)2D3 was inefficient in amelioration of established liver fibrosis. Administration of 1,25(OH)2D3 to bile duct ligation rats led to a high mortality rate probably caused by hypercalcemia. We conclude that 1,25(OH)2D3 may be considered as a potential preventive treatment in an in vivo model but failed to ameliorate established cirrhosis.

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