Optimizacija i primjena metode inhibicije hijaluronidaze za određivanje učinka lista masline (Olea europaea, L.)

master's thesis

• Title (english): Optimization of hyaluronidase inhibition method and determination of inhibition by olive leaf extracts (Olea europaea, L.)
• Author: Ana Kadi
• Mentor: Marijana Zovko Končić (mentor)
• Committee member: Marijana Zovko Končić (predsjednik povjerenstva)
• Committee member: Maja Bival Štefan (član povjerenstva)
• Committee member: Jasna Jablan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmacognosy); Zagreb
• Defense date and country: 2022-07-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Hijaluronska kiselina jedna je od esencijalnih komponenata tkiva koja sudjeluje u održavanju hidratacije, međustaničnoj komunikaciji i djeluje kao fiziološki lubrikant. Hijaluronidaza, kao enzim koji razgrađuje hijaluronsku kiselinu, sve više je u fokusu znanstvenih istraživanja u svrhu prevencije preuranjenog starenja kože i usporavanja progresije pojedinih bolesti. Cilj ovog diplomskog rada bila je optimizacija spektrofotometrijske metode inhibicije hijaluronidaze i njezina primjena na ekstraktima lista masline. U eksperimentima je korištena hijaluronidaza iz goveđih testisa i natrijeva sol hijaluronske kiseline. Metoda je prilagođena laboratorijskim uvjetima i primijenjena na ekstraktima lista masline te je ispitan njihov inhibicijski potencijal za enzim hijaluronidazu. Pozitivna kontrola bila je taninska kiselina, no najniži IC50, odnosno ukupno najjaču inhibiciju ostvarilo je otapalo B (IC50=3,47 ± 1,45 μg/mL; sastav: 28,8% PPG, 63,8% destilirana voda, 7,4% mliječna kiselina). Najsnažniju inhibiciju među ekstraktima pokazao je uzorak B042M, s IC50 vrijednošću od 4,91 ± 0,56 μg/mL. IC50 vrijednosti čistih otapala (A i B) usporedive su s IC50 vrijednostima dobivenima za ekstrakte priređene s pripadajućim otapalima, stoga se može zaključiti kako sama otapala najviše utječu na izmjerenu inhibiciju. Za potvrdu i točnije definiranje inhibicijskog učinka sastavnica iz lista masline potrebno je provesti dodatna istraživanja.

Abstract (english)

Hyaluronic acid is an essential component of tissues and some of its roles are retaining hydration, intercellular communication, and lubrication. Hyaluronidase, as an enzyme that degrades hyaluronic acid, attracts the attention of researchers who study premature skin aging and different diseases connected to disbalanced hyaluronan homeostasis. The goal of this diploma thesis is to optimize the spectrophotometric method for determining hyaluronidase inhibition. Bovine testicular hyaluronidase was the experimental enzyme and sodium hyaluronate was its substrate. The method has been adjusted to laboratory requirements and was used for measuring hyaluronidase - inhibiting potential of olive leaf extracts. Tannic acid was used as positive control, but the lowest IC50 and overall strongest inhibition was exhibited by solvent B (IC50=3,47 ± 1,45 μg/mL; 28,8% PPG, 63,8% purified water, 7,4% lactic acid). Sample B042M exhibited the strongest inhibition of all the extracts, with IC50 of 4.91 ± 0.56 μg/mL. IC50 values of pure solvents (A and B) is comparable to the values measured for the extracts. This result indicates that the solvents alone act as strong inhibitors of the hyaluronidase enzyme. For exact defining of inhibition by olive leaf extracts, more research needs to be done.

• Keywords: hijaluronidaza; hijaluronska kiselina; list masline; inhibicija
• Keywords (english): hyaluronidase; hyaluronic acid; olive leaf extract; inhibition
• Language: croatian
• URN:NBN: urn:nbn:hr:163:097233
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2024-07-07
• Created on: 2022-07-26 12:55:01