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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I28-R145-HWA_5916 |
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dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-145 |
| collection |
Repositorio Digital de la Universidad de Buenos Aires (UBA) |
| language |
Español |
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spa |
| topic |
Porfiria aguda intermitente (PAI) d-aminolevulinato sintetasa-1 (ALA-S1) Suero fetal bovino (SFB) Factor de crecimiento similar a la insulina tipo 1 (IGF-1 Acute Intermittent Porphyria (AIP) Delta-aminolevulinic synthase 1 Fetal Bovine Serum (FBS) Insulin like growth factor (IGF1) Ciencias de la vida |
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Porfiria aguda intermitente (PAI) d-aminolevulinato sintetasa-1 (ALA-S1) Suero fetal bovino (SFB) Factor de crecimiento similar a la insulina tipo 1 (IGF-1 Acute Intermittent Porphyria (AIP) Delta-aminolevulinic synthase 1 Fetal Bovine Serum (FBS) Insulin like growth factor (IGF1) Ciencias de la vida Mora León, Sandra Milena Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| topic_facet |
Porfiria aguda intermitente (PAI) d-aminolevulinato sintetasa-1 (ALA-S1) Suero fetal bovino (SFB) Factor de crecimiento similar a la insulina tipo 1 (IGF-1 Acute Intermittent Porphyria (AIP) Delta-aminolevulinic synthase 1 Fetal Bovine Serum (FBS) Insulin like growth factor (IGF1) Ciencias de la vida |
| description |
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. |
| author2 |
Gerez, Esther Noemí |
| author_facet |
Gerez, Esther Noemí Mora León, Sandra Milena |
| format |
Tesis de maestría Tesis de maestría acceptedVersion |
| author |
Mora León, Sandra Milena |
| author_sort |
Mora León, Sandra Milena |
| title |
Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| title_short |
Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| title_full |
Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| title_fullStr |
Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| title_full_unstemmed |
Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa |
| title_sort |
efecto del factor de crecimiento similar a la insulina tipo 1 (igf-1) sobre la d-aminolevulinato sintetasa |
| publisher |
Facultad de Farmacia y Bioquímica |
| publishDate |
2018 |
| url |
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 |
| work_keys_str_mv |
AT moraleonsandramilena efectodelfactordecrecimientosimilaralainsulinatipo1igf1sobreladaminolevulinatosintetasa |
| _version_ |
1766017553441751040 |
| spelling |
I28-R145-HWA_59162022-04-12 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. Fil: Mora León, Sandra Milena. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Gerez, Esther Noemí Facultad de Farmacia y Bioquímica Rossetti, María Victoria Mora León, Sandra Milena 2018-03-13 La porfiria aguda intermitente (PAI) se caracteriza por una disminución en la actividad de la enzima porfobilinógeno deaminasa, asociada a un marcado incremento en la expresión de la primera enzima del camino del hemo, la d-aminolevulinato sintetasa 1 (ALA-S1) y una acumulación de los precursores neurotóxicos ALA y PBG.\nEl suero fetal bovino (SFB) contiene un gran número de factores macromoleculares y nutricionales esenciales para el crecimiento celular, entre ellos pequeñas moléculas como aminoácidos, azúcares, lípidos y hormonas como el factor de crecimiento similar a la insulina tipo 1 (IGF-1) o somatomedina, similar en estructura molecular a la insulina. Es así como, IGF-1 en altas concentraciones puede interactuar con los receptores de insulina, ejerciendo efectos insulino miméticos. La insulina actúa sobre los factores de transcripción que regulan los niveles de ARNm del ALA-S1, a través de la vía PI3K/Akt.\nDado estos antecedentes, el objetivo de este estudio fue determinar el efecto de IGF-1 sobre la regulación del ALA-S1.\nEn la primera parte de los ensayos in vitro se trabajó con células C3A incubadas en medio DMEM y tratadas con diferentes concentraciones de SFB durante 2hs, lo que provocó un aumento en los niveles de proteína del ALA-S1. Con el fin de corroborar si este incremento fue debido una mayor transcripción, se decidió cuantificar los niveles de ARNm por la técnica de real time. Contrariamente a lo esperado se notó una disminución de éstos con respecto a los controles. Para evaluar si los cambios eran a nivel traduccional, se utilizó en los ensayos SFB + cicloheximida (CHX). Los resultados permitieron determinar que el aumento observado en el contenido de proteína del ALA-S1 por el SFB es debido a una mayor traducción.\nEn la segunda parte, las células C3A fueron tratadas con diferentes concentraciones (1, 10, 50 nM) de IGF-1 durante 15 minutos en presencia y\nii\nausencia de AG1024/L, un inhibidor selectivo de la fosforilación de tirosina de la subunidad ? del receptor de IGF-1. Los controles fueron tratados con el vehículo correspondiente.\nSe observó que el tratamiento con IGF-1 provocó una caída en los niveles basales de ARNm del ALA-S1 y este efecto fue revertido por la presencia de AG1024/L.\nEstos resultados nos permiten sugerir que IGF-1 participaría en la expresión basal del ALA-S1. application/pdf De Siervi, Adriana Jawerbaum, Alicia Randi, Andrea Porfiria aguda intermitente (PAI) d-aminolevulinato sintetasa-1 (ALA-S1) Suero fetal bovino (SFB) Factor de crecimiento similar a la insulina tipo 1 (IGF-1 Acute Intermittent Porphyria (AIP) Delta-aminolevulinic synthase 1 Fetal Bovine Serum (FBS) Insulin like growth factor (IGF1) spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Efecto del factor de crecimiento similar a la insulina tipo 1 (IGF-1) sobre la d-aminolevulinato sintetasa info:eu-repo/semantics/masterThesis info:ar-repo/semantics/tesis de maestría info:eu-repo/semantics/acceptedVersion 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 |