The ketogenic diet (KD) is a diet with low level of carbohydrates, a good amount of protein and high-fat, established over the years as a nonpharmacologic treatment option used to ameliorate and cure various disease, including principally epilepsy, thanks to the fundamental action of d-β-hydroxybutyrate (BHB), a natural ketone body produced during a period of carbohydrate deprivation used as an energy source for the body cells, in particular for heart and brain. Among the fat compound of KD, there are some fat-soluble vitamins with an antioxidant action such as Vitamin E, a compound also available in organic food products, which could contribute to this marked protective effect. The aim of the doctoral project was to evaluate the anti- inflammatory and antioxidant effects of the KD associated with the addition of biological products on the central nervous system (CNS). BV2 cells culture, microglial cells representing the first line of brain immune system defence, as first experimental phase was treated with 100uM of Vitamin E to evaluate its neuroprotective effects through MTT assay, morphological and migration tests, ELISA test, Western blotting analysis and qRT-PCR in the presence or absence of proinflammatory stimuli such as lipopolysaccharide (LPS). In addition, as second experimental phase, to confirm the neuroprotective effect of KD, the objective was to evaluate the possible anti-inflammatory effect of BHB, in a concentration of 5mM on BV2 cells, chosen for MTT test morphological and migration assay, with or without the LPS and finally for ELISA test. The results affirmed that Vitamin E and BHB have an anti-inflammatory and antioxidant action in this context, both can polarize the microglia towards the anti-inflammatory phenotype, showing the ability to influence cell migration and modulate anti-inflammatory pathway. In this context one could postulate that BHB could be a key molecule in the prevention of neurodegenerative diseases and therefore, since BHB is a product of KD, one could indirectly provide evidence for the potential role of KD in the processes of neuroinflammation and neuroprotection. Furthermore, Vitamin E also seems to have potential and effectiveness in this context, hypothesizing its use in the prevention and management of neurodegenerative diseases, suggesting its possible use as a food supplement. Naturally, although the results are encouraging, further studies are needed to clarify the molecular mechanisms involved.
La dieta chetogenica (KD) è una dieta a basso contenuto di carboidrati, un buon apporto di proteine e un alto contenuto di grassi, affermatasi negli anni come opzione terapeutica non farmacologica utilizzata per migliorare e curare diverse malattie, tra cui principalmente l'epilessia, grazie alla azione fondamentale del d- β-idrossibutirrato (BHB), un corpo chetonico naturale prodotto durante un periodo di deprivazione dei carboidrati usato come fonte di energia per le cellule dell'organismo, in particolare per cuore e cervello. Tra i composti lipidici della KD ci sono alcune vitamine liposolubili ad azione antiossidante come la Vitamina E, composto disponibile anche nei prodotti alimentari biologici, che potrebbe contribuire a questo marcato effetto protettivo. Lo scopo del progetto di dottorato è stato di valutare gli effetti antinfiammatori e antiossidanti della KD associata all'aggiunta di prodotti biologici sul sistema nervoso centrale (SNC). La coltura cellulare BV2, cellule microgliali che rappresentano la prima linea di difesa del sistema immunitario cerebrale, come prima fase sperimentale, sono state trattate con 100uM di Vitamina E per valutarne i suoi effetti neuroprotettivi mediante il saggio MTT, test morfologici e di migrazione, test ELISA, analisi di Western blotting e qRT- PCR in presenza o assenza di stimoli pro-infiammatori come il Lipopolisaccaride (LPS). Inoltre, come seconda fase sperimentale, per confermare l'effetto neuroprotettivo della KD, l'obiettivo è stato quello di valutare il possibile effetto antinfiammatorio del BHB, in una concentrazione di 5 mM, sulle cellule BV2, scelta per il test morfologico e di migrazione del MTT, con o senza LPS, ed infine per il test ELISA. I risultati hanno affermato che, in questo contesto, la Vitamina E ed il BHB hanno un’azione antinfiammatoria e antiossidante, entrambi possono polarizzare la microglia verso il fenotipo antinfiammatorio, mostrando la capacità di influenzare la migrazione cellulare e modulare le vie di segnalazione antinfiammatorie. In questo contesto si potrebbe postulare che il BHB potrebbe essere una molecola chiave nella prevenzione delle malattie neurodegenerative e quindi, poiché il BHB è un prodotto della KD, si potrebbe indirettamente fornire prova del potenziale ruolo della KD nei processi di neuro-infiammazione e neuro-protezione. Inoltre, anche la Vitamina E sembra avere potenzialità ed efficacia in questo contesto, ipotizzando un suo utilizzo nella prevenzione e gestione delle malattie neurodegenerative, suggerendone un possibile utilizzo come integratore alimentare. Naturalmente, anche se i risultati sono incoraggianti, sono necessari ulteriori studi per chiarire i meccanismi molecolari coinvolti.
Antioxidant and anti-inflammatory activity of the ketogenic diet with enhancement of the use of organic food products / LA TORRE, MARIA ESTER. - (2024). [10.14274/la-torre-maria-ester_phd2024]
Antioxidant and anti-inflammatory activity of the ketogenic diet with enhancement of the use of organic food products
LA TORRE, MARIA ESTER
2024-01-01
Abstract
The ketogenic diet (KD) is a diet with low level of carbohydrates, a good amount of protein and high-fat, established over the years as a nonpharmacologic treatment option used to ameliorate and cure various disease, including principally epilepsy, thanks to the fundamental action of d-β-hydroxybutyrate (BHB), a natural ketone body produced during a period of carbohydrate deprivation used as an energy source for the body cells, in particular for heart and brain. Among the fat compound of KD, there are some fat-soluble vitamins with an antioxidant action such as Vitamin E, a compound also available in organic food products, which could contribute to this marked protective effect. The aim of the doctoral project was to evaluate the anti- inflammatory and antioxidant effects of the KD associated with the addition of biological products on the central nervous system (CNS). BV2 cells culture, microglial cells representing the first line of brain immune system defence, as first experimental phase was treated with 100uM of Vitamin E to evaluate its neuroprotective effects through MTT assay, morphological and migration tests, ELISA test, Western blotting analysis and qRT-PCR in the presence or absence of proinflammatory stimuli such as lipopolysaccharide (LPS). In addition, as second experimental phase, to confirm the neuroprotective effect of KD, the objective was to evaluate the possible anti-inflammatory effect of BHB, in a concentration of 5mM on BV2 cells, chosen for MTT test morphological and migration assay, with or without the LPS and finally for ELISA test. The results affirmed that Vitamin E and BHB have an anti-inflammatory and antioxidant action in this context, both can polarize the microglia towards the anti-inflammatory phenotype, showing the ability to influence cell migration and modulate anti-inflammatory pathway. In this context one could postulate that BHB could be a key molecule in the prevention of neurodegenerative diseases and therefore, since BHB is a product of KD, one could indirectly provide evidence for the potential role of KD in the processes of neuroinflammation and neuroprotection. Furthermore, Vitamin E also seems to have potential and effectiveness in this context, hypothesizing its use in the prevention and management of neurodegenerative diseases, suggesting its possible use as a food supplement. Naturally, although the results are encouraging, further studies are needed to clarify the molecular mechanisms involved.File | Dimensione | Formato | |
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