Biološki učinci fenolnih kiselina iz odabranih vrsta porodice Lamiaceae

doctoral thesis

• Title (english): Biological effects of phenolic acids from selected Lamiaceae species
• Author: Maja Bival Štefan; MBZ: 264244
• Mentor: Sanda Vladimir-Knežević (mentor)
• Committee member: Renata Jurišić Grubešić (član povjerenstva)
• Committee member: Biljana Blažeković (član povjerenstva)
• Committee member: Donatella Verbanac (član povjerenstva); MBZ: 133213
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmacognosy); Zagreb
• Defense date and country: 2015-07-23, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy
• Universal decimal classification (UDC): 615 - Pharmacology. Therapeutics. Toxicology

Abstract

U okviru ovog doktorskog rada po prvi put je provedeno opsežno usporedno istraživanje bioloških učinaka
14 prirodnih fenolnih kiselina, prevalidacija metode određivanja ukupnih hidroksicimetnih derivata u biljnim
drogama, fitokemijska karakterizacija fenolnih kiselina i flavonoida te biološki učinci odabranih vrsta roda
Micromeria Bentham (Lamiaceae). Antioksidacijski učinak fenolnih kiselina ispitan je primjenom četiri različite
spektrofotometrijske metode. Hidroksicimetne kiseline su pokazale bolja antioksidacijska svojstva u odnosu na
većinu hidroksibenzojevih kiselina, a iznimno dobar antioksidacijski učinak utvrđen je za ružmarinsku, kavenu i
galnu kiselinu. Citotoksični učinak fenolnih kiselina ispitan je na HepG2 i THP 1 staničnim kulturama primjenom
MTS testa, pri čemu većina testiranih kiselina nije pokazala značajnije smanjenje vijabilnosti stanica. Istraživanje
protuupalnog djelovanja fenolnih kiselina istaknulo je galnu, ružmarinsku, klorogensku, kavenu i sinapinsku kiselinu
kao dobre inhibitore interleukina-6. Komet testom na uzorku humane krvi nakon tretiranja bovericinom, dokazan je
dobar antigenotoksični učinak svih ispitanih fenolnih kiselina.Fitokemijska analiza fenolnih kiselina i flavonoida u
biljnim vrstama porodice Lamiaceae provedena je primjenom kromatografskih (HPTLC, RP-HPLC-DAD, LC-ESIMS/
MS) i spektrofotometrijskih metoda. U etanolnim ekstraktima vrsta M. croatica, M. juliana i M. thymifolia
identificirano je jedanaest fenolnih kiselina i petnaest flavonoida. Kriptoklorogenska, neoklorogenska, 3-p- i 4-pkumaroilkina,
3-(3,4- dihidroksifenil)mliječna, salvianolna kiselina B i J te dimer i trimer ružmarinske kiseline po
prvi put su identificirane u navedenim vrstama. Izuzev luteolinskih i akacetinskih derivata u vrstama M. juliana i M.
thymifolia, ostali su flavonoidi također prvi put identificirani u ispitivanim vrstama roda Micromeria. Ustanovljena su
dobra antioksidacijska, protuupalna i antigenotoksična svojstva odabranih biljnih vrsta, pri čemu se istaknula
endemična vrsta M. croatica.
Dobiveni rezultati ukazali su na veliki biomedicinski potencijal ispitanih fenolnih kiselina i vrsta roda Micromeria.

Abstract (english)

A comprehensive comparative study of the biological effects of 14 natural phenolic acids, prevalidation of
total hydroxycinnamic assays, identification of phenolic acids and flavonoids as well as biological activities of
selected Micromeria species were conducted for the first time in this doctoral thesis.
Antioxidant activity of phenolic acids was tested using four different spectrophotometric methods. Hydroxycinnamic
acids had better antioxidant properties than most hydroxybenzoic acids. Evaluation of antioxidant activity highlighted
rosmarinic, caffeic and gallic acids as the best ones. Cytotoxic effect of phenolic acids was studied on HepG2 and
THP-1 cell cultures using MTS test, and phenolic acids did not show significant reduction of cell viability.
Investigation of phenolic acids' anti-inflammatory activity revealed gallic, rosmarinic, chlorogenic, caffeic and
sinapinic acids as good inhibitors of interleukin-6 production. Comet assay on human blood cells treated with
beauvericin proved good antigenotoxic effects of all tested phenolic acids.
Phytochemical analysis of polyphenols was carried out using various chromatographic (HPTLC, RP-HPLC-DAD, LCESI-
MS/MS) and spectrophotometric methods. Eleven phenolic acids and fifteen flavonoids were identified in the
ethanolic extracts of M. croatica, M. juliana and M. thymifolia. Chriptochlorogenic, neochlorogenic, 3-p- and 4-pcoumaroylquinic,
3-(3,4- dihydroxyphenil)lactic, salvianolic acid B and J as well as dimer and trimer of rosmarinic
acid were identified in these species for the first time. With exception of luteolin and acacetin derivatives in M. juliana
and M. thymifolia, flavonoids were identified for the first time in the tested species. Good antioxidant, antiinflammatory
and antigenotoxic properties were established for investigated Micromeria species with special emphasis
on endemic M. croatica.
Obtained results revealed the great biomedical potential of tested phenolic acids and Micromeria species.

• Keywords: fenolne kiseline; flavonoidi; Micromeria; antioksidacijski učinak; citotoksični učinak; protuupalni učinak; IL-6; antigenotoksični učinak
• Keywords (english): phenolic acids; flavonoids; Micromeria; antioxidant effect; cytotoxic effect; antiinflammatory effect; IL-6; antigenotoxic effect
• Language: croatian
• URN:NBN: urn:nbn:hr:163:698085
• Study programme: Title: Pharmacy and biochemistry; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor znanosti (doktor znanosti)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2016-04-06 13:44:11