Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa
Intermittent acute porphyria (PAI) is characterized by a decrease in the activity of the enzyme Porphobilinogen deaminase, associated with a significantincrease in the expression of the first heme pathway enzyme, d-aminolevulinate synthetase 1 (ALA-S1) and an accumulation of neurotoxic precursors AL...
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| Formato: | Tesis de maestría acceptedVersion |
| Lenguaje: | Español |
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Facultad de Farmacia y Bioquímica
2018
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| Acceso en línea: | http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_5916 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_5916.dir/5916.PDF |
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| Sumario: | Intermittent acute porphyria (PAI) is characterized by a decrease in the activity of the enzyme Porphobilinogen deaminase, associated with a significantincrease in the expression of the first heme pathway enzyme, d-aminolevulinate synthetase 1 (ALA-S1) and an accumulation of neurotoxic precursors ALA and PBG.\nFetal bovine serum (FBS) contains a large number of macromolecular and nutritional factors which are essential for cell growth, including small molecules such as amino acids, sugars, lipids and hormones such as insulin-like growth factor-1 (IGF-1) or somatomedin, whose molecular structure is similar to insulin. Thus, IGF-1 at high concentrations can interact with insulin receptors, exerting insulin mimicking effects. Insulin acts on the transcription factors that regulate ALA-S1 mRNA levels, via the PI3K / Akt pathway.\nThe objective of this project is to establish the in vitro conditions to study the effect of IGF-1 on the regulation of ALA-S1.\nIn the first part of the tests in vitro we worked with cellular line C3A. Cells were incubated in DMEM medium and treated with different concentrations of FBS for 2hs, which led to an increase in protein levels of ALA-S1. In order to evaluate if this increase was caused by a higher transcription, mRNA levels were quantified by Real Time analysis. Contrary to expectations, there was a decrease of mRNA levels with respect to the controls. To evaluate whether the changes were at the translational level, SFB + cycloheximide (CHX) were used in the assays. The results allowed to determine that the observed increase in the protein content of ALA-S1 by SFB was caused by an increased translation.\nIn the second part of the project, C3A cells were fasted for 18 hrs and then treated with different concentrations (1, 10, 50 nM) of IGF-1 for 15 min in the presence and absence of AG1024/L, a selective inhibitor of tyrosine\niv\nphosphorylation of IGF-1 receptor ?-subunit. Controls were treated with the corresponding vehicle.\nIGF-1 treatment caused a decrease of basal ALA-S1 mRNA levels and this effect was reversed by the presence of AG1024/L.\nThese results allow us to suggest that IGF-1 would participate in the basal expression of ALA-S1. |
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