CASTELLANO, LORENZO (2025) PRECLINICAL CELLULAR MODELS OF PARKINSON’S DISEASE: AN USEFUL TOOL FOR THE STUDY OF NEUROPROTECTIVE STRATEGIES. [Tesi di dottorato]

Documento PDF TESI_FINALE_LORENZO_CASTELLAN_CICLO_37_PON.pdf Visibile a [TBR] Amministratori dell'archivio Download (3MB) | Richiedi una copia

Abstract

The cellular and molecular mechanisms underlying motor and non-motor symptoms in Parkinson’s disease (PD) encompass a range of pathological processes, including alterations in the conformational structure of synaptic proteins, mitochondrial dysfunction, oxidative stress, and metabolic imbalances (Goyal & Chaturvedi, 2021; Adolphs, 2009; Christidi et al., 2018; Bora et al., 2015). These factors contribute to the intricate dynamics of neuroplasticity modifications within the context of progressive neurodegeneration, which is closely linked to the cognitive reserve of PD patients at both early and advanced disease stages (Petrosini et al., 2009; Rosemberg et al., 2018). In this framework, pathological accumulation of alpha-synuclein and mitochondrial dysfunction emerge as critical contributors to cellular impairment in PD, given their causal relationship with disease progression.This study has been performed to evaluate the neuroprotective effects of citicoline, L-carnitine and glutathione in in vitro models reproducing sporadic and familial forms of PD in order to verify the hypothesis that pharmacological strategies potentially able to modulate mitochondrial function and pathological protein aggregation might be useful to slowdown the neurodegeneration occurring in PD.In this regard, the study has been performed in vitro, in SH-SY5Y neuroblastoma-derived cells exposed to rotenone treatment, as a model of sporadic PD, and in human SH-SY5Y cells stably transfected with the WT or the human mutation A53T of α-synuclein, as a model of familial PD, to investigate the effects of the above mentioned molecules on the cellular metabolism and mitochondrial function and to correlate these effects with the modifications of α-synuclein aggregation possibly revealed following treatment with the molecules under examination. Therefore, the first experiments performed were addressed to evaluate the mitochondrial redox activity in SH-SY5Y cells exposed to 500nM rotenone, an inhibitor of mitochondrial Complex I, for 24 hr, and in cells stably transfected with alpha-synuclein WT and containing the human mutation A53T. The results of these experiments demonstrated that in both experimental conditions the mitochondrial oxidative capacity was impaired. Afterward, time course and dose-response experiments have been performed in untransfected SH-SY5Y cells, exposed to citicoline, L-carnitine and glutathione, in order to identify the appropriate concentrations and times of exposure for each of the tested molecules able to affect mitochondrial oxidative capacity without induce any cellular toxicity.The results obtained demonstrated that the decrease in redox activity occurring in SH-SY5Y cells exposed to rotenone treatment for 24 hr was prevented by the treatment with L-carnitine, citicoline and glutathione. Similarly, in stably transfected SH-SY5Y cells the impairment in mitochondrial redox activity was observed in A53T-alpha-synuclein-transfected cells, an effect that was prevented by L-carnitine, citicoline and glutathione exposure.

Downloads

Actions (login required)

Modifica documento