Selekcija biološki aktivnih tvari sa zaštitnim učincima na citotoksičnost induciranu polikloriranim bifenilima

doctoral thesis

• Title (english): Selection of biologically active compounds with protective effects on cytotoxicity induced by polychlorinated biphenyls
• Author: Marina Miletić
• Mentor: Ivana Kmetič (mentor)
• Committee member: Teuta Murati (predsjednik povjerenstva)
• Committee member: Kristina Radošević (član povjerenstva)
• Committee member: Jelka Pleadin (član povjerenstva)
• Committee member: Veronika Kovač (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Toxicology); Zagreb
• Defense date and country: 2022-02-18, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology
• Universal decimal classification (UDC): 57 - Biological sciences. Physical anthropology. Biodiversity

Abstract

Poliklorirani bifenili (engl. polychlorinated biphenyls, PCB) su perzistentni organski zagađivači okoliša koji pokazuju mnogobrojne negativne učinke na okoliš te na zdravlje ljudi (oksidacijski stres, inflamatorni učinak, endokrina modulacija, mutagenost). U ovom istraživanju su primjenom in vitro test sustava ispitani potencijalni zaštitni učinci dva odabrana polifenola – resveratrola (RSV) i kurkumina (CRC), kao i ferocenskih analoga polifenola (ferocenski triacilni derivat resveratrola – RF i ferocenski analog kurkumina – CF) na citotoksičnost induciranu odabranim PCB kongenerima – PCB 77 („dioksinu sličan“, planarni tetraklorobifenil) i PCB 153 („ne-dioksinu sličan“, neplanarni heksaklorobifenil). Prethodno su na stanicama ovarija kineskog hrčka (CHO-K1) i kanceroznoj hepatocelularnoj staničnoj liniji (Hepa1-6) utvrđene doze polifenola i njihovih analoga koje ne djeluju toksično na stanice: 2,5 – 20 μM RSV; 1 – 20 μM CRC; 2,5 – 100 μM RF i 1 – 20 μM CF. Za oba PCB kongenera (75 μM) utvrđen je statistički značajan inhibitorni učinak na vijabilnost CHO-K1 stanica, a izraženiji citotoksični učinak određen je za PCB 77. Zaštitni učinci RSV, CRC, RF i CF iskazani su prvenstveno na citotoksičnost induciranu PCB-em 153. Primjenom in vitro metoda najizraženiji zaštitni učinak na citotoksičnost induciranu PCB-em 153 utvrđen je za RF i to u svim ispitanim koncentracijama, dok su pozitivni učinci RSV-a i CRC-a utvrđeni samo pri najnižim ispitanim dozama (≤ 2,5 μM). Zaštitni učinak RSV-a na citotoksičnost induciranu PCB-em 77 nije utvrđen niti jednom od primijenjenih metoda, već je određen sinergistički antiproliferativni učinak RSV-a i planarnog kongenera PCB-a. Analiza stanične smrti metodom protočne citometrije pokazala je da predinkubacija RSV-om i CRC-om djeluje zaštitno pri izloženosti CHO-K1 stanica ortho-supstituiranom kongeneru, dok je pri izloženosti planarnom kongeneru određen sinergistički proapoptotski učinak što je u korelaciji s primijenjenim metodama praćenja vijabilnosti. Oba analoga polifenola poboljšala su vijabilnost kulture i pokazala antiapoptotski učinak pri tretmanu PCB-em 153, za razliku od citotoksičnosti inducirane PCB-em 77, gdje je samo CF pokazao zaštitni učinak. Usporedbom učinaka svih ispitanih spojeva (RSV, CRC, RF, CF) na oksidacijski stres induciran PCB-ima, utvrđeno je da RF ima najizraženije antioksidacijsko djelovanje (u dozama 1 – 20 μM) pri tretmanu stanica PCB-em 77.

Abstract (english)

Polychlorinated biphenyls (PCBs) are persistent organic pollutants that show numerous adverse effects on the environment and human health (oxidative stress, inflammatory effects, endocrine modulation, mutagenicity). In this study, the potential protective effects of two selected polyphenols - resveratrol (RSV) and curcumin (CRC), as well as their ferrocene analogues (ferrocene triacyl derivative of resveratrol - RF and ferrocene analogue of curcumin - CF), on cytotoxicity induced by selected PCB congeners - PCB 77 ("dioxin-like", planar tetrachlorobiphenyl) and PCB 153 ("non-dioxin-like", non-planar hexachlorobiphenyl) were studied using an in vitro test systems. Previously, doses of polyphenols and their analogues, which have no toxic effects on cells, were determined on Chinese hamster ovary cells (CHO-K1) and on the hepatocellular carcinoma cell line (Hepa1-6): 2.5 - 20 μM RSV; 1 - 20 μM CRC; 2.5 - 100 μM RF and 1 - 20 μM CF. A statistically significant inhibitory effect on CHO-K1 cell viability was found for both PCB congeners (75 μM), with a more pronounced cytotoxic effect determined for PCB 77. The protective effects of RSV, CRC, RF and CF were most evident in PCB 153-induced cytotoxicity. Using in vitro methods, the most pronounced protective effect on PCB 153 induced cytotoxicity was found for RF at all concentrations tested, whereas the beneficial effects of RSV and CRC were found only at the lowest doses tested (≤ 2.5 μM). The protective effect of RSV on PCB 77-induced cytotoxicity was not detected by any of the methods used, but a synergistic antiproliferative effect of RSV and planar PCB congener was determined. Analysis of cell death by flow cytometry showed that preincubation with RSV and CRC had a protective effect on exposure of CHO-K1 cells to ortho-substituted congener, whereas exposure to planar congener was found to have a synergistic proapoptotic effect that correlated with the viability monitoring methods used. Both polyphenol analogues improved culture viability and showed an antiapoptotic effect in PCB 153-treated cells, in contrast to PCB 77-induced cytotoxicity where only CF showed a protective effect. Comparing the effects of all tested compounds (RSV, CRC, RF, CF) on PCB-induced oxidative stress, RF was found to have the most pronounced antioxidant effect (doses 1 - 20 μM) in PCB 77-treated cells.

• Keywords: resveratrol; kurkumin; ferocenski analozi polifenola; PCB 77; PCB 153; in vitro test sustavi; citotoksičnost; stanična smrt; oksidacijski stres
• Keywords (english): resveratrol; curcumin; ferrocene analogues of polyphenols; PCB 77; PCB 153; in vitro test systems; cytotoxicity; cell death; oxidative stress
• Language: croatian
• URN:NBN: urn:nbn:hr:159:523418
• Promotion: 2022
• Study programme: Title: Postgraduate PhD Study in Nutrition; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje biotehničkih znanosti, polje nutricionizam (doktor/doktorica znanosti, područje biotehničkih znanosti, polje nutricionizam)
• Type of resource: Text
• Extent: 206 str. ; 30 cm
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-11-09 11:49:31