Marco, Dacrema (2022) NUTRACEUTICAL TO PREVENT URINARY TRACT INFECTIONS: PRECLINICAL STUDIES. [Tesi di dottorato]

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Abstract

In the first and second year of my PhD course the aim was to find a new non-antibiotic candidate of plant origin, which could be use in the management of the UTIs. In the first phase, I selected five different plants, which were extracted to explore their potential against uropathogenic E. coli. According to the literature the plants selected for the studies were: Camellia sinensis L. (Leaves), Punica Granatum L. (Fruit), Orthosiphon stamineus (Leaves), Morus alba L. (Leaves), H. Sabdariffa (Flowers). All the plants were extracted, and the extracts screened to select the most promising plant extract according to their polyphenol concentration, antioxidant, and antimicrobial activities. Based on these results we selected only one of the most promising extracts (H. Sabdariffa) and we performed UHPLC coupled to IT-TOF profile assessing the main compounds and understanding the possible mechanism of action of these molecules against the uropathogenic E. coli. Finally, there was the evaluation anti-invasivity activity using bladder carcinoma cells (HT-1376). In the third year of the PhD course, which took place at the Leibniz-Institut für Lebensmittel-Systembiologie an der Technischen Universität München, a non-canonical approach was followed, with the final aim to update the literature on the TAS2R expression and the in vitro bioavailability of Cucurbitacin B, which is contained in Momordica charantia L. fruits. For this reason M. charantia. fruits were divided into seeds, pulp, and skin and these parts and the whole fruit were extracted three consecutive times using water, acetonitrile/water (70/30) and ethyl acetate. The extracts were characterized with an untargeted UHPLC-Tof-MS screening. After the metabolic profile assessment and the determination of cucurbitacins in the extracts, the next step was understood if cucurbitacin B, present in each part of the fruit, was able to reach the GU tract using the transwell system. Moreover, considering the poor literature on TAS2R in CACO-2 cell line we decided also to explore their expression in this specific cell line and to treat the cells with bitter melon extract to understand if the treatment could change the expression of this receptor.

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