CPEB4 Increases Expression of PFKFB3 to Induce Glycolysis and Activate Mouse and Human Hepatic Stellate Cells, Promoting Liver Fibrosis
| Autor: | Anna Manzano, Salvador Naranjo-Suarez, Marta Ramirez, Raúl Méndez, Mercedes Fernandez, Javier Gallego, Clara Suñer, Marc Mejias, Ramon Bartrons, Nuria Pell |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Liver Cirrhosis Male Small interfering RNA Platelet-derived growth factor Phosphofructokinase-2 Primary Cell Culture Liver Cirrhosis Experimental Liver cells Cell Line Small hairpin RNA Cèl·lules hepàtiques 03 medical and health sciences chemistry.chemical_compound Mice 0302 clinical medicine Fibrosis medicine Hepatic Stellate Cells Animals Humans Carbon Tetrachloride Liver diseases Mice Knockout Gene knockdown Hepatology biology Malalties del fetge Gastroenterology RNA-Binding Proteins Transforming growth factor beta medicine.disease Cell biology Rats Up-Regulation 030104 developmental biology Glucòlisi chemistry Gene Expression Regulation Liver Gene Knockdown Techniques Hepatic stellate cell biology.protein 030211 gastroenterology & hepatology Cell activation Glycolysis |
| Zdroj: | Dipòsit Digital de la UB Universidad de Barcelona |
| Popis: | BACKGROUND & AIMS: We investigated mechanisms of hepatic stellate cell (HSC) activation, which contributes to liver fibrogenesis. We aimed to determine whether activated HSCs increase glycolysis, which is regulated by 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3), and whether this pathway might serve as a therapeutic target. METHODS: We performed studies with primary mouse HSCs, human LX2 HSCs, human cirrhotic liver tissues, rats and mice with liver fibrosis (due to bile duct ligation [BDL] or administration of carbon tetrachlo- ride), and CPEB4-knockout mice. Glycolysis was inhibited in cells and mice by administration of a small molecule antagonist of PFKFB3 (3-[3-pyridinyl]-1-[4-pyridinyl]-2- propen-1-one [3PO]). Cells were transfected with small interfering RNAs that knock down PFKFB3 or CPEB4. RESULTS: Up-regulation of PFKFB3 protein and increased glycolysis were early and sustained events during HSC activation and accompanied by increased expression of markers of fibrogenesis; incubation of HSCs with 3PO or knockdown of PFKFB3 reduced their activation and prolif- eration. Mice with liver fibrosis after BDL had increased hepatic PFKFB3; injection of 3PO immediately after the surgery prevented HSC activation and reduced the severity of liver fibrosis compared with mice given vehicle. Levels of PFKFB3 protein were increased in fibrotic liver tissues from patients compared with non-fibrotic liver. Up-regulation of PFKFB3 in activated HSCs did not occur via increased transcription, but instead via binding of CPEB4 to cyto- plasmic polyadenylation elements within the 3'-untranslated regions of PFKFB3 messenger RNA. Knockdown of CPEB4 in LX2 HSCs prevented PFKFB3 overexpression and cell acti- vation. Livers from CPEB4-knockout had decreased PFKFB3 and fibrosis after BDL or administration of carbon tetra- chloride compared with wild-type mice. CONCLUSIONS: Fibrotic liver tissues from patients and rodents (mice and rats) have increased levels of PFKFB3 and glycolysis, which are essential for activation of HSCs. Increased expression of PFKFB3 is mediated by binding of CPEB4 to its untranslated messenger RNA. Inhibition or knockdown of CPEB4 or PFKFB3 prevents HSC activation and fibrogenesis in livers of mice. |
| Databáze: | OpenAIRE |
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