Hemijska karakterizacija, antimikrobna i citotoksična aktivnost etarskog ulja i postdestilacionog ostatka kleke (Juniperius communis L.) i njihovo kombinovano delovanje sa konvencionalnim antibioticima i citostaticima
Chemical characterization, antimicrobial and cytotoxic activity of essential oil and post-distillation waste of juniper (Juniperus communis L.) and their combinations with conventional antibiotics and cytostatics
Докторанд
Vasilijević, BojanaМентор
Nikolić, BiljanaЧланови комисије
Knežević-Vukčević, JelenaMitić-Ćulafić, Dragana
Orčić, Dejan
Srdić-Rajić, Tatjana
Метаподаци
Приказ свих података о дисертацијиСажетак
U ovom radu ispitivana su etarska ulja i postdestilacioni ostatak (PDO) Juniperus communis
L. GC-MS analiza ulja je pokazala dominantno učešće α-pinena, a LC-MS/MS je identifikovala
hinsku kiselinu i rutin kao najzastupljenije u PDO. Bolja antioksidativna aktivnost PDO utvrđena je
u TBA i naročito DPPH eseju (IC50 4,4 μg/mL), što potvrđuju i rezultati FRAP testa. Citotoksičnost
je bila veća u slučaju ulja: IC50 vrednosti na ćelijskim linijama HT-29, HCT 116, A549 i MRC-5
bile su u opsegu 69-120 μg/mL. Efekat kombinacija oba derivata sa doksorubicinom na kancerskim
A549 i normalnim MRC-5 ćelijama bio je sinergistički, naročito u slučaju A549. Citometrijska
analiza i alkalni komet test su pokazali da PDO indukuje apoptozu, zaustavlja ćelijski ciklus i ima
genotoksični efekat.
MIK testom je utvrđeno da su Listeria monocytogenes i Aspergillus versicolor najosetljivije,
posebno prema PDO (MIK 0,39 i 0,12 mg/mL). Sinergizam je uočen u slučaju tretmana listerije
kombinacijama ulja/PDO i strep...tomicina/ampicilina/azitromicina, kao i Candida albicans
kombinacijom PDO i bifonazola. Adhezija patogena na ćelije kolona u kulturi bila je inhibirana
dejstvom PDO ili ulja, u slučaju listerije ili kandide. Etarska ulja kleke i čubra, primenjena u
mikrodilucionim testovima odvojeno ili u kombinacijama, pokazala su dobar potencijal i postojanje
sinergizma u slučaju primoizolata L. monocytogenes. Time kill test je potvrdio sinergizam i ukazao
na veću osetljivost listerije u aerobnim nego hipoksičnim uslovima. Ulja primenjena na
mariniranom mesu inhibirala su rast ukupnih aerobnih heterotrofa, enterobakterija, bakterija mlečne
kiseline i inokulisane listerije.
Uočeni citotoksični i antimikrobni efekat preporučuju derivate kleke za adjuvante koji
pojačavaju aktivnost konvencionalnih lekova.
In this work essential oils and post-distillation waste (PDW) of Juniperus communis L. were
screened. GC-MS analysis pointed at α-pinene as dominant constituent of oils; LC-MS/MS
identified quinic acid and rutin as the most prevalent in PDW. Higher antioxidativity of PDW was
shown in TBA and especially DPPH assay (IC50 4.4 μg/mL), which is additionally confirmed by
FRAP test. Oil induced higher cytotoxicity against HT-29, HCT 116, A549 and MRC-5 cells, with
IC50 values ranging 69-120 μg/mL. Combinations of each derivative with doxorubicin, applied on
cancer A549 and normal MRC-5 cells induced synergism especially pronounced in A549.
Cytometric analysis and alkaline comet test showed that PDW induced apoptosis and cell cycle
arrest, and was genotoxic.
Microdilution assay pointed at Listeria monocytogenes and Aspergillus versicolor as the
most sensitive, especially to PDW (MIC 0.39 and 0.12 mg/mL). Synergism was observed in the
case of listeria treated with combinations of oil/PDW and st...reptomycin/ampicillin/azithromycin, as
well as Candida albicans with a combination of PDW and bifonazole. In vitro adhesion of L.
monocytogenes/C. albicans to colon cells was inhibited by PDW and oil, respectively. Additionaly,
juniper and savory essential oils, applied in microdilution tests separately or in combinations,
showed notable potential and synergism in the case of primoisolates of listeria. Time kill assay
confirmed synergism and indicated a higher bacterial sensitivity under aerobic than hypoxic
conditions. Oils applied to marinated meat inhibited the growth of total aerobic heterotrophs,
enterobacteria, lactic acid bacteria and inoculated listeria.
Observed cytotoxic and antimicrobial effect recommends juniper derivatives as adjuvants
that enhance conventional drugs activity.
Факултет:
Универзитет у Београду, Биолошки факултетДатум одбране:
27-02-2021Пројекти:
- Биолошки активни природни производи као потенцијални извори нових лекова и дијететских суплемената (RS-MESTD-Basic Research (BR or ON)-172058)