|Tipo di tesi||Tesi di laurea magistrale|
|Titolo||AZIONE DEI NEUROSTEROIDI SU NEURONI E MICROGLIA IN UN MODELLO CELLULARE DI MORBO DI PARKINSON|
|Titolo in inglese||ACTIONS OF NEUROSTEROIDS ON NEURONS AND MICROGLIA IN A CELLULAR MODEL OF PARKINSON'S DESEASE|
|Struttura||Dipartimento di Scienze della Vita|
|Corso di studi||CHIMICA E TECNOLOGIA FARMACEUTICHE (D.M. 270/04)|
|Data inizio appello||2019-04-02|
|Disponibilità||Accessibile via web (tutti i file della tesi sono accessibili)|
Il Morbo di Parkinson (PD) è una malattia neurodegenerativa nella quale si ha una morte selettiva dei neuroni dopaminergici della Substantia Nigra (SN). I meccanismi responsabili della degenerazione neuronale dopaminergici sono sconosciuti; tuttavia la principale ipotesi è quella neuroinfiammatoria. Nella genesi della malattia hanno un ruolo importante non solo i neuroni ma anche le cellule gliali; infatti composti che riducono l'attivazione gliale proteggono la SN in modelli in vivo di PD. I neurosteroidi (Ns) hanno effetti neuroprotettivi ed i loro livelli cerebrali variano in molte patologie degenerative.
Parkinson's disease (PD) is a neurodegenerative syndrome in which there is a selective death of the dopaminergic neurons of Substantia Nigra (SN). The mechanisms responsible for neuronal dopaminergic degeneration are unknown; however the main hypothesis is the neuroinflammatory one. Not only neurons but also glial cells play an important role in the genesis of the disease; in fact compounds that reduce glial activation protect the SN in in vivo models of PD. Neurosteroids (Ns) have neuroprotective effects and their brain levels fluctuate in many degenerative diseases. We used an in vitro PD model that consists in treating a neuronal and a microglial cell line, with a toxin: Rotenone. These cultures were treated with Allopregnanolone (ALLO), Dehydroepiandrosterone (DHEA) and Pregnenolone sulfate (PGLS). We measured cell viability and markers of microglial activation (free oxygen radicals and nitric oxide) under several experimental conditions Understanding the influence of neurosteroids on communication between neurons and microglia to maintain homeostasis and CNS integrity may have important implications for the therapeutic potential of neurosteroids in neurodegenerative diseases such as PD. The objective of the project is to confirm the aforementioned hypothesis and to investigate the molecular mechanism underlying the neuroprotective effect of neurosteroids. The use of co-cultures of neurons and microglia allows evaluating the "cross talk" between these two cell types in neurodegenerative conditions and the possible involvement of Ns. This will permit deepening some of the cellular mechanisms responsible for a neurodegenerative pathology such as PD in order to find new alternatives of pharmacological intervention. Using an in vitro PD model, consisting in the treatment of a neuronal cell line (SH-SY5Y; SH) and a microglial (BV-2) with the Rotenone toxin (Rot at concentrations between 0.01 and 50 uM), we measured cell viability and cytotoxicity (using the MTT and LDH tests, respectively) in our experimental conditions. Since this study represents the first part of a broader and more innovative research project, the preliminary phases were dedicated to the development and optimization of methods and experiments. The results indicated that Rotenone, when administered alone, causes SH-SY5Y and BV-2 cell death but with different potencies: BV-2 cells were more sensitive to toxin than SH. The evaluated neurosteroids differentially improve the viability of both cell types; also in this case the effect is evident at different concentrations: depending on the cell type. The toxic effect of Rot is significantly reduced after co-application with Allo even if, in BV-2 cells, it is not completely abolished. Our data therefore seem to support the hypothesis that neurosteroids can exert a neuroprotective effect on SH-SY5Y cells In conclusion, the data obtained allowed to shed light, at least in part, on some mechanisms involved in Rot toxicity in distinct cell types. Further studies are needed to clarify the molecular mechanisms underlying these effects, aimed at expanding the knowledge of Parkinson's disease and the rational use of neurosteroids in this pathology.