Liver fibrosis is characterized by the activation of perivascular hepatic stellate cells (HSCs), the release of fibrogenic nanosized extracellular vesicles (EVs), and increased HSC glycolysis. Nevertheless, how glycolysis in HSCs coordinates fibrosis amplification through tissue zone-specific pathways remains elusive. Here, we demonstrate that HSC-specific genetic inhibition of glycolysis reduced liver fibrosis. Moreover, spatial transcriptomics revealed a fibrosis-mediated up-regulation of EV-related pathways in the liver pericentral zone, which was abrogated by glycolysis genetic inhibition. Mechanistically, glycolysis in HSCs up-regulated the expression of EV-related genes such as Ras-related protein Rab-31 (RAB31) by enhancing histone 3 lysine 9 acetylation on the promoter region, which increased EV release. Functionally, these glycolysis-dependent EVs increased fibrotic gene expression in recipient HSC. Furthermore, EVs derived from glycolysis-deficient mice abrogated liver fibrosis amplification in contrast to glycolysis-competent mouse EVs. In summary, glycolysis in HSCs amplifies liver fibrosis by promoting fibrogenic EV release in the hepatic pericentral zone, which represents a potential therapeutic target.
- MeSH
- extracelulární vezikuly * metabolismus MeSH
- glykolýza * MeSH
- jaterní cirhóza * metabolismus patologie genetika MeSH
- jaterní hvězdicovité buňky * metabolismus patologie MeSH
- játra metabolismus patologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- Rab proteiny vázající GTP metabolismus genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND & AIMS: Growth Differentiation Factor 11 (GDF11) is an anti-aging factor, yet its role in liver diseases is not established. We evaluated the role of GDF11 in healthy conditions and in the transition from non-alcoholic fatty liver disease (NAFLD) to non-alcoholic steatohepatitis (NASH). RESULTS: GDF11 mRNA levels positively correlated with NAFLD activity score and with CPT1, SREBP, PPARγ and Col1A1 mRNA levels, and associated to portal fibrosis, in morbidly obese patients with NAFLD/NASH. GDF11-treated mice showed mildly exacerbated hepatic collagen deposition, accompanied by weight loss and without changes in liver steatosis or inflammation. GDF11 triggered ALK5-dependent SMAD2/3 nuclear translocation and the pro-fibrogenic activation of HSC. CONCLUSIONS: GDF11 supplementation promotes mild liver fibrosis. Even considering its beneficial metabolic effects, caution should be taken when considering therapeutics that regulate GDF11. METHODS: We analyzed liver biopsies from a cohort of 33 morbidly obese adults with NAFLD/NASH. We determined the correlations in mRNA expression levels between GDF11 and genes involved in NAFLD-to-NASH progression and with pathological features. We also exposed wild type or obese mice with NAFLD to recombinant GDF11 by daily intra-peritoneal injection and monitor the hepatic pathological changes. Finally, we analyzed GDF11-activated signaling pathways in hepatic stellate cells (HSC).
- MeSH
- buněčné linie MeSH
- dospělí MeSH
- experimentální cirhóza jater chemicky indukované metabolismus patologie MeSH
- jaterní cirhóza diagnóza etiologie genetika metabolismus MeSH
- jaterní hvězdicovité buňky metabolismus patologie MeSH
- játra metabolismus patologie MeSH
- kostní morfogenetické proteiny genetika metabolismus toxicita MeSH
- lidé středního věku MeSH
- lidé MeSH
- morbidní obezita komplikace diagnóza MeSH
- myši inbrední C57BL MeSH
- nealkoholová steatóza jater diagnóza etiologie genetika metabolismus MeSH
- progrese nemoci MeSH
- růstové diferenciační faktory genetika metabolismus toxicita MeSH
- signální transdukce MeSH
- studie případů a kontrol MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hepatocellular carcinoma (HCC) has a poor outcome. Most HCCs develop in the context of liver fibrosis and cirrhosis caused by chronic inflammation. Short-term fasting approaches enhance the activity of chemotherapy in preclinical cancer models, other than HCC. Multi-tyrosine kinase inhibitor Sorafenib is the mainstay of treatment in HCC. However, its benefit is frequently short-lived. Whether fasting can alleviate liver fibrosis and whether combining fasting with Sorafenib is beneficial remains unknown. A 24 hr fasting (2% serum, 0.1% glucose)-induced changes on human hepatic stellate cells (HSC) LX-2 proliferation/viability/cell cycle were assessed by MTT and flow cytometry. Expression of lypolysaccharide (LPS)-induced activation markers (vimentin, αSMA) was evaluated by qPCR and immunoblotting. Liver fibrosis and inflammation were evaluated in a mouse model of steatohepatitis exposed to cycles of fasting, by histological and biochemical analyses. A 24 hr fasting-induced changes were also analyzed on the proliferation/viability/glucose uptake of human HCC cells exposed to Sorafenib. An expression panel of genes involved in survival, inflammation, and metabolism was examined by qPCR in HCC cells exposed to fasting and/or Sorafenib. Fasting decreased the proliferation and the activation of HSC. Repeated cycles of short term starvation were safe in mice but did not improve fibrosis. Fasting synergized with Sorafenib in hampering HCC cell growth and glucose uptake. Finally, fasting normalized the expression levels of genes which are commonly altered by Sorafenib in HCC cells. Fasting or fasting-mimicking diet diets should be evaluated in preclinical studies as a mean to potentiate the activity of Sorafenib in clinical use.
- MeSH
- buňky Hep G2 MeSH
- časové faktory MeSH
- experimentální cirhóza jater metabolismus patologie MeSH
- fenylmočovinové sloučeniny farmakologie MeSH
- glukosa metabolismus MeSH
- hepatocelulární karcinom farmakoterapie genetika metabolismus patologie MeSH
- jaterní hvězdicovité buňky účinky léků metabolismus patologie MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- myši inbrední C57BL MeSH
- nádory jater farmakoterapie genetika metabolismus patologie MeSH
- nealkoholová steatóza jater metabolismus patologie MeSH
- niacinamid analogy a deriváty farmakologie MeSH
- omezení příjmu potravy metabolismus MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky farmakologie MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- alkoholická cirhóza jater diagnóza prevence a kontrola terapie MeSH
- biliární cirhóza diagnóza etiologie terapie MeSH
- chronická nemoc MeSH
- ezofageální a žaludeční varixy etiologie prevence a kontrola terapie MeSH
- hepatitida C terapie MeSH
- hepatitida etiologie klasifikace komplikace MeSH
- jaterní cirhóza * diagnóza etiologie terapie MeSH
- jaterní hvězdicovité buňky cytologie patologie MeSH
- lidé MeSH
- myofibroblasty fyziologie patologie účinky léků MeSH
- nemoci jater * etiologie klasifikace terapie MeSH
- portální hypertenze etiologie prevence a kontrola terapie MeSH
- prognóza MeSH
- regenerace jater * fyziologie imunologie účinky léků MeSH
- ztučnělá játra diagnóza etiologie terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Activated hepatic stellate cells (HSC) are a major source offibrous proteins in cirrhotic liver. Inducing or accelerating their apoptosis is a potential way of liver fibrosis treatment. Extracellular matrix (ECM) surrounding cells in tissue affects their differentiation, migration, proliferation and function. Type I collagen is the main ECM component in fibrotic liver. We have examined how this protein modifies apoptosis of normal rat HSC induced by gliotoxin, cycloheximide and cytochalasin D in vitro and spontaneous apoptosis of HSC isolated from CCl4-damaged liver. We have found that type I collagen gel enhances HSC apoptosis regardless of the agent triggering this process.
- MeSH
- apoptóza účinky léků MeSH
- buněčné kultury MeSH
- chlorid uhličitý MeSH
- cykloheximid MeSH
- cytochalasin D MeSH
- gliotoxin MeSH
- jaterní cirhóza patologie MeSH
- jaterní hvězdicovité buňky účinky léků patologie MeSH
- kolagen typu I farmakologie MeSH
- krysa rodu rattus MeSH
- modely nemocí na zvířatech MeSH
- potkani Sprague-Dawley MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The signalling pathway elicited by hepatocyte growth factor (HGF) and its receptor c-Met is indispensable for liver development and regeneration. It has been described that c-Met is released from the cell surface by a disintegrin and metalloprotease 10 (ADAM10) resulting in a soluble c-Met form known as sMet. Using the human hepatocellular HepG2 and hepatic stellate cell LX2 lines we show that sMet is released from the cell surface of liver cells by both ADAM17 and ADAM10, with ADAM17 appearing to be the major proteinase. Moreover, using a mouse model of 3,5-diethoxycarbonyl- 1,4-dihydroxycollidine (DDC)-induced hepatobiliary obstruction we show that serum levels of sMet correlate well with the liver damage state and consecutive regeneration as well as with established markers of liver damage such as alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bilirubin. However, sMet exhibited remarkably better correlation with liver damage and inflammation than did serum tumour necrosis factor α (TNF-α), whose shedding is also mediated by ADAM proteolytic activity. Our results indicate that the proteolytic activity of ADAM10/17 is essential for regulating HGF/c-Met signalling during acute liver damage and following regeneration and that the differential serum levels of sMet together with expression of c-Met/HGF might be a useful indicator not only for damage, but also for ongoing liver regeneration.
- MeSH
- alanintransaminasa krev MeSH
- aspartátaminotransferasy krev MeSH
- bilirubin metabolismus MeSH
- biologické markery metabolismus MeSH
- buňky Hep G2 MeSH
- hepatocyty metabolismus patologie MeSH
- jaterní hvězdicovité buňky metabolismus patologie MeSH
- játra metabolismus patologie MeSH
- lidé MeSH
- membránové proteiny metabolismus MeSH
- messenger RNA genetika metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nemoci jater krev metabolismus patologie MeSH
- proteiny ADAM metabolismus MeSH
- protoonkogenní proteiny c-met krev metabolismus MeSH
- rozpustnost MeSH
- sekretasy metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Jaterní fibróza představuje významný proces v progresi chronických jaterních onemocnění. V procesu fibrogeneze mají prvořadou roli hvězdicové buňky a komponenty extracelulární matrix. „Zlatým standardem“ v diagnostice jaterní fibrózy je biopsie a histologické vyšetření. Neinvazívní diagnostické postupy zahrnují vyšetření sérových markerů fibrózy, zobrazovací metody – ultrazvuk, magnetická rezonance, měření tuhosti jater FibroScanem nebo MR elastografií a dechové testy. Neinvazívní postupy jsou přínosné především v diagnostice rozvinuté fibrózy a cirhózy.
Liver fibrosis represents an important process in progression of chronic liver diseases. Stellate cells and compoments of extracellular matrix play the principal role in the process of fibrogenesis. Liver biopsy and histological examination are considered to be the “gold standard” in diagnosis of liver fibrosis. Noninvasive diagnostic procedures include examination of serum markers of fibrosis, imaging methods include ultrasound, magnetic resonance, assessment of liver stiffness with FibroScan or MR elastography; breath tests can be used as well. Noninvasive procedures are useful especially in diagnosis of advanced fibrosis and cirrhosis.
- Klíčová slova
- hvězdicové buňky, sesérové markeryrum markers, elastrografie,
- MeSH
- biologické markery analýza krev MeSH
- biopsie metody MeSH
- dechové testy metody přístrojové vybavení MeSH
- elastografie metody přístrojové vybavení MeSH
- histologické techniky metody MeSH
- jaterní cirhóza diagnóza etiologie patologie MeSH
- jaterní hvězdicovité buňky patologie MeSH
- lidé MeSH
- sérum enzymologie chemie MeSH
- ultrasonografie metody MeSH
- Check Tag
- lidé MeSH
