|Tipo di tesi||Tesi di laurea magistrale|
|Titolo||Determinazione dei ROS in modelli cellulari e animali di Morbo di Parkinson e valutazione del possibile ruolo protettivo di estratti di frutti rossi|
|Titolo in inglese||Determination of ROS in cellular and animal models of Parkinson's disease and evaluation of the possible protective role of red fruit extracts|
|Struttura||Dipartimento di Scienze della Vita|
|Corso di studi||CHIMICA E TECNOLOGIA FARMACEUTICHE (D.M. 270/04)|
|Data inizio appello||2021-12-20|
|Disponibilità||Embargo di 3 anni|
|Data di rilascio||2024-12-20|
Il Morbo di Parkinson (MP) è la seconda malattia neurodegenerativa più diffusa al mondo, caratterizzata dalla morte precoce dei neuroni dopaminergici della Substantia Nigra pars compacta, regione del cervello responsabile del controllo del movimento e della coordinazione.
Parkinson's disease (PD) is the second most common neurodegenerative disease in the world, characterized by the early death of dopaminergic neurons in the Substantia Nigra pars compacta, the brain region responsible for controlling movement and coordination. Although the causes of the disease are not currently known, various studies have shown that one of the factors responsible for neurodegeneration is oxidative stress, which also occurs following an excessive production of oxygen free radicals (ROS) and/or due to poor efficiency of antioxidant systems. To date, an effective treatment has not yet been identified in countering the evolution of PD and inhibiting neurodegeneration and recently, products of natural origin with antioxidant properties are becoming important as complementary therapy. Polyphenols, important secondary metabolites of plants, have antioxidant and anti-inflammatory properties. They could inhibit the formation of alpha-synuclein aggregates (Lewy bodies) and reduce oxidative stress. Among the fruits rich in polyphenols, attention was focused on extracts of bilberry: Vaccinium myrtillus L., and sweet cherry "Moretta di Vignola": Prunus avium L. They have high concentrations of anthocyanins, polyphenols potentially capable of chelate metals and eliminate free radicals produced in abnormal quantities in individuals with PD. In the present study, using two techniques - flow cytometry and plate fluorimetry - and two different fluorochromes - CellROX Green Reagent (CGR) and H2DCFDA - ROS production was evaluated on neuroblastoma cell lines (SH-SY5Y) and microglia (BV2) treated with Rotenone (ROT) - neurotoxin of plant origin capable of inducing blockage of the mitochondrial respiratory chain - and / or with extracts of bilberry and Moretta di Vignola. Experiments were carried out in which conditioned culture medium of BV2 microglial cells treated with extracts and / or ROT were added to SH-SY5Y cells to evaluate their possible influence in the production of free radicals. ROS production was also evaluated in vivo on adult male heads of Drosophila melanogaster (D.) - animal model of MP - treated with ROT and / or Moretta extract. Comparing the two techniques, plate fluorimetry provided greater reproducibility and reliability, and H2DCFDA showed good labeling unlike CGR which, under our conditions, is unusable. The not always effective staining and other criticalities observed with CGR in flow cytometry could be overcome by using H2DCFDA. The results obtained in vitro suggest a potential antioxidant effect of the extracts, which are shown to be effective in countering the ROT-induced ROS production on both cell types. Even the conditioned medium of microglia treated with Moretta can contrast the production of ROS in SH-SY5Y cells suggesting a possible interaction between the two cell lines. For the dosage of ROS in the Drosophila experiments, fluorimetric analysis with H2DCFDA was used. The results obtained suggest a potential antioxidant effect of the extract that seems effective even in vivo in opposing the production of ROS induced by Rotenone.