Utjecaj otapala na vijabilnost ljudskih stanica i određivanje parametara kinetike enzima acetilkolinesteraze

undergraduate thesis

• Title (english): The effects of solvents on the viability of human cells and the determination of the kinetic parameters of the enzyme acetylcholinesterase
• Author: Gabrijela Zibar
• Mentor: Igor Stuparević (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2023-09-08, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

U ovom radu ispitivan je utjecaj otapala dimetil sulfoksida (DMSO) i etanola (EtOH) na aktivnost ljudske acetilkolinesteraze i vijabilnost PC-3 stanica raka prostate s ciljem određivanja koncentracija otapala pri kojima bi se mogla provoditi medicinska istraživanja lijekova bez značajnog utjecaja otapala na rezultate ispitivanja. Aktivnost acetilkolinesteraze u prisutnosti otapala određivana je metodom po Ellmanu te se iz dobivenih rezultata može zaključiti kako koncentracija DMSO do 0,1 % te EtOH do 0,5 % nema značajan inhibicijski učinak na enzim, pri koncentracijama supstrata acetiltiokolina od 0,1, 0,2 i 0,4 mmol/dm³ . Pri ispitivanju citotoksičnosti otapala na PC-3 stanicama koristila se MTS metoda te je iz dobivenih rezultata vidljivo kako pri koncentraciji DMSO od 0,625 % ne dolazi do značajnog gubitka vijabilnosti, dok za EtOH ta koncentracija iznosi 5 %. Iz sveukupnih rezultata vidljivo je kako DMSO ima jači inhibicijski i citotoksični učinak, iz čega se može zaključiti kako bi pri ispitivanju učinkovitosti lijekova EtOH bio bolji izbor kao otapalo.

Abstract (english)

In this study, the impact of the solvents dimethyl sulfoxide (DMSO) and ethanol (EtOH) on the activity of human acetylcholinesterase and the viability of PC-3 prostate cancer cells was examined, with the aim of determining solvent concentrations at which medicinal drug research could be conducted without significant solvent interference on the test results. The activity of acetylcholinesterase in the presence of solvents was determined using the Ellman method, and from the obtained results, it can be concluded that a DMSO concentration of up to 0,1 % and an EtOH concentration of up to 0,5 % do not have a significant inhibitory effect on the enzyme at substrate concentrations of acetylthiocholine
of 0,1, 0,2 and 0,4 mmol/dm³. In the investigation of solvent cytotoxicity on PC-3 cells, the MTS method was used, and from the obtained results, it is evident that at a DMSO concentration of 0,625 % there is no significant loss of viability, whereas for EtOH, that concentration is 5 %. Overall, the results indicate that DMSO has a stronger inhibitory and cytotoxic effect, leading to the conclusion that EtOH would be a better choice as a solvent in drug efficacy studies.

• Keywords: dimetil sulfoksid; etanol; acetilkolinesteraza; PC-3 stanice
• Keywords (english): dimethyl sulfoxide; ethanol; acetylcholinesterase; PC-3 cells
• Language: croatian
• URN:NBN: urn:nbn:hr:159:038687
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-04-18 10:49:49