Effects of lipopolysaccharide and abscisic acid on insulin signaling in SH-SY5Y cell line
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Título
Effects of lipopolysaccharide and abscisic acid on insulin signaling in SH-SY5Y cell lineAutoría
Tutor/Supervisor; Universidad.Departamento
Sánchez Pérez, Ana Maria; Universitat Jaume I. Unitat Predepartamental de MedicinaFecha de publicación
2021-07-22Editor
Universitat Jaume IResumen
Insulin resistance is a pathophysiological mechanism closely related to neuroinflammation and
present in many neurodegenerative diseases like Alzheimer's Disease. Under pathological
conditions, abnormal increased ... [+]
Insulin resistance is a pathophysiological mechanism closely related to neuroinflammation and
present in many neurodegenerative diseases like Alzheimer's Disease. Under pathological
conditions, abnormal increased phosphorylation of insulin receptor substrate 1 (IRS1) serine
residues occurs, leading the disruption of the phosphoinositol 3-kinase/protein kinase B
(PI3K/Akt) pathway and resulting in neuronal impairment. Lipopolysaccharide (LPS) is a proinflammatory molecule present in the outer cell wall of Gram-negative bacteria, which is
recognized by microglial and neuronal tall-like receptors, thus leading neuroinflammation.
Abscisic acid (ABA) is a sesquiterpene that has shown protective effects against peripheral insulin
resistance, neuroinflammation and cognitive impairment. Besides, ABA has shown
differentiation properties in glioblastoma cell line. In this study, we explored the effects of in vitro
LPS and ABA administration on insulin signaling in neurons, acutely and chronically. In addition,
we tested ABA as a neuronal differentiating stimulus on a neuroblastoma cell line. For that,
functional assays were performed on the SH-SY5Y human neuroblastoma cell line. On the one
hand, we pre-treated the cells with ABA (20 μM) and 20 hours later we administrated LPS
(1μg/mL) acutely (24h). On the other hand, we treated cells chronically with LPS (1μg/mL) and
ABA (20 μM) simultaneously for 5 days. Then, cells were treated with insulin (1 or 10 mU/mL)
for 5 minutes and cell lysis was performed. Finally, we analyzed the phosphorylation of insulin
signaling key proteins such as IRS1pTyr, IRS1pSer616, pAkt and pERK1/2 by western blot.
Moreover, we analyzed qualitatively ABA and retinoic acid (RA, positive control) as neuronal
differentiation stimuli in SH-SY5Y cell line by inverted optical microscope images. As a result,
the comparison of groups (mean ± SEM) treated with LPS, ABA, ABA+LPS and negative control
showed no significant differences in the phosphorylation of IRS1pTyr, IRS1pSer616, pAkt and
pERk1/2 after insulin stimulation. On the other side, RA-treated neuroblastoma cells showed a
differentiated phenotype, while ABA did not stimulate differentiation. Therefore, our data
suggested that LPS did not affect insulin signaling, both acutely and chronically. Since there were
no differences in terms of LPS administration, the neuroprotective effects of ABA against LPSinduced neuroinflammation could not be ascertained. Moreover, ABA did not display a neuronal
differentiating stimulus on the human neuroblastoma cell line SH-SY5. [-]
Palabras clave / Materias
Descripción
Treball Final de Màster Universitari en Investigació en Cervell i Conducta. Codi: SBM024. Curs: 2020/2021.
Tipo de documento
info:eu-repo/semantics/masterThesisDerechos de acceso
info:eu-repo/semantics/openAccess