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Assessment of activity and expression of selected isoforms of glutathione S-transferase in the in vivo model of glutamate-induced obesity
Košťáková, Šárka ; Boušová, Iva (advisor) ; Skálová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Šárka Košťáková Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Assessment of activity and expression of selected isoforms of glutathione S-transferase in the in vivo model of glutamate-induced obesity Obesity is a chronic disease with an increasing prevalence worldwide. This disease is associated with many pathophysiological changes in the organism; a possible influence on the antioxidant defenses of the organism is one of them. The aim of this study was to determine specific activity, protein expression and mRNA level of selected isoforms of glutathione S-transferase (GST) in the cytosol obtained from kidney and heart of the male NMRI mice with the obesity induced by repeated subcutaneous administration of monosodium glutamate (MSG) and in healthy controls. The specific activities were assayed by spectrophotometric method using universal and specific substrates. Protein expression of GST isoforms from GSTA, GSTM and GSTP families was monitored using denaturing polyacrylamide gel electrophoresis and immunoblotting. The mRNA level of selected GST genes was determined by real-time PCR. In the cytosolic fraction of MSG mice kidney, increase in total GST specific...

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