Biološka aktivnost hidrokinona i nitrofurantoina i njihov utjecaj na proteomski profil stanične linije mokraćnoga mjehura T24

doctoral thesis

• Title (english): Biological activity of hydroquinone and nitrofurantoin and their influence on the proteomic profile of the T24 bladder cell line
• Author: Ana Huđek Turković
• Mentor: Ksenija Durgo (mentor)
• Mentor: Gordana Rusak (komentor)
• Committee member: Draženka Komes (predsjednik povjerenstva)
• Committee member: Nevenka Kopjar (član povjerenstva)
• Committee member: Ana Butorac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2023, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology
• Universal decimal classification (UDC): 578/579 - Virology. Microbiology

Abstract

U ovom radu provedena je kvalitativna i kvantitativna analiza vodenog ekstrakta lišća medvjetke (Arctostaphylos uva-ursi L.) koji se često samoinicijativno koristi u zajedničkoj terapiji s antibiotikom nitrofurantoinom za liječenje urinarnih infekcija. Ukupni polifenolni sastav i antioksidacijski potencijal ekstrakta određeni su spektofotometrijski, a pojedinačni spojevi arbutin i hidrokinon tekućinskom kromatografijom visoke djelotvornosti (HPLC). Metodom simulirane in vitro probave određena je stabilnost uzoraka. Biološki učinci ekstrakta istraženi su spektrofotometrijskim metodama na straničnim linijama probavnog sustava (CAL 27, AGS, Caco-2) i jetre (HepG2) u uvjetima in vitro. Stanična linija karcinoma mokraćnog mjehura (T24) korištena je kao modelni sustav ispitivanja citotoksičnog, prooksidacijskog i genotoksičnog učinka hidrokinona, nitrofurantoina i njihove smjese pomoću metoda Neutral Red, DCFH-DA i mikronukleus cytome testa. Također, razlika u ekspresiji proteina u T24 stanicama nakon tretmana spomenutim spojevima, određena je spektrometrijom masa. Glavni rezultati ovog rada pokazuju sinergističko djelovanje hidrokinona i antibiotika nitrofurantoina na T24 stanice mokraćnog mjehura u vidu smanjenja preživljenja, povećanja proooksidacijskog učinka i genotoksičnog djelovanja te promjene ekspresije proteina. Prema tome, može se zaključiti da biljne spojeve treba tretirati kao kemijske spojeve koji, ovisno o uvjetima, mogu pokazati sinergistička, antagonistička i potencirajuća svojstva u kombinaciji s drugim terapeuticima te negativno utjecati na ljudsko zdravlje.

Abstract (english)

In this study, the aqueous extract of bearberry leaves (Arctostaphylos uva-ursi L.), which is frequently self-administered in combination therapy with the antibiotic nitrofurantoin for the treatment of urinary infections, was subjected to a qualitative and quantitative analysis. The total polyphenol composition and antioxidant potential of the extract were determined spectrophotometrically, and the individual compounds arbutin and hydroquinone were determined by high-performance liquid chromatography (HPLC). The in vitro biological effect of the extract on the cell lines of the gastrointestinal tract (CAL 27, AGS, Caco-2) and liver (HepG2) were assessed by spectrophotometric techniques. In vitro digestion was used to determine the stability of the extract during intake. Using the Neutral Red, DCFH-DA, and micronucleus cytome assays, the bladder cancer cell line (T24) was utilised as a model system to examine the cytotoxic, prooxidative, and genotoxic effects of hydroquinone, nitrofurantoin, and their combination. Additionally, mass spectrometry was used to identify the differences in protein expression in T24 cells following treatment with the abovementioned substances. The major findings of this study demonstrate the synergistic effects of hydroquinone and the antibiotic nitrofurantoin on human urinary bladder cancer cells (T24), including decreased survival, increased pro-oxidative effect and genotoxic effect, and altered protein expression. It follows that herbal compounds should be viewed as chemical substances that, depending on how they are used, may exhibit synergistic, antagonistic, and potentiating effects when combined with other therapies, and may have a negative influence on human health.

• Keywords: vodeni ekstrakt lišća medvjetke; hidrokinon; nitrofurantoin; smjesa spojeva; in vitro; biološko djelovanje
• Keywords (english): aqueous bearberry leaf extract; hydroquinone; nitrofurantoin; mixture of compounds; in vitro; biological activity
• Language: croatian
• URN:NBN: urn:nbn:hr:159:914920
• Promotion: 2023
• Study programme: Title: Postgraduate PhD Study in Nutrition; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/ doktorica znanosti, područje biotehničkih znanosti, odgovarajuće polje (određeno matičnom strukom) (doktor/ doktorica znanosti, područje biotehničkih znanosti, odgovarajuće polje (određeno matičnom strukom))
• Type of resource: Text
• Extent: 184 str. ; 30 cm
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-10-25 09:51:13