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2016-02

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Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/167757

Chitosan-mediated shRNA knockdown of cytosolic alanine aminotransferase improves hepatic carbohydrate metabolism

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https://doi.org/10.1007/s10126-015-9670-8

Resum

Alanine aminotransferase (ALT) catalyses a transamination reaction that links carbohydrate and amino acid metabolism. In this study, we examined the effect of silencing cytosolic ALT (cALT) expression on the hepatic metabolism in Sparus aurata. A number of siRNA and shRNA designed to down-regulate cALT expression were validated in HEK-293 cells transfected with plasmids expressing S. aurata cALT or mitochondrial ALT (mALT) isoforms: ALT silencing significantly decreased the expression levels of S. aurata mRNA cALT1 to 62 % (siRNA) and 48 % (shRNA) of the values observed in control cells. The effect of cALT silencing was analysed in the liver of S. aurata 72 h after intraperitoneal injection of chitosan-tripolyphosphate (TPP) nanoparticles complexed with a plasmid encoding a shRNA to down-regulate cALT expression (pCpG-si1sh1). In fish fed diets with different ratio of protein to carbohydrate and treated with chitosan-TPP-pCpG-si1sh1, cALT1 and cALT2 mRNA levels significantly decreased irrespective of the diet. Consistently, ALT activity decreased in liver of treated animals. In the liver of S. aurata treated with chitosan-TPP-pCpG-si1sh1 nanoparticles, down-regulation of cALT expression increased the activity of key enzymes in glycolysis (6-phosphofructo-1-kinase and pyruvate kinase) and protein metabolism (glutamate dehydrogenase). Besides showing for the first time that administration of chitosan-TPP-pCpG-si1sh1 nanoparticles silences hepatic cALT expression in vivo, our data support that down-regulation of cALT could improve the use of dietary carbohydrates to obtain energy and spare protein catabolism

Matèries

Quitosan, Teràpia genètica, Orada, Nanopartícules, Metabolisme

Matèries (anglès)

Chitosan, Gene therapy, Sparus aurata, Nanoparticles, Metabolism

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Articles publicats en revistes (Bioquímica i Fisiologia)
Articles publicats en revistes (Farmàcia, Tecnologia Farmacèutica i Fisicoquímica)

Citació

GONZÁLEZ, Juan d., SILVA-MARRERO, Jonás i., METÓN TEIJEIRO, Isidoro, CABALLERO SOLARES, Albert, VIEGAS, Iván, FERNÁNDEZ GONZÁLEZ, Felipe javier, MIÑARRO CARMONA, Montserrat, FÁBREGAS, A. (anna), TICÓ GRAU, Josep r., JONES, John g., VÁZQUEZ BAANANTE, Ma. isabel. Chitosan-mediated shRNA knockdown of cytosolic alanine aminotransferase improves hepatic carbohydrate metabolism. _Marine Biotechnology_. 2016. Vol. 18, núm. 1, pàgs. 85-97. [consulta: 23 de gener de 2026]. ISSN: 1436-2228. [Disponible a: https://hdl.handle.net/2445/167757]

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