Ispitivanje genotoksičnih i citotoksičnih efekata vrsta Artemisia vulgaris i Artemisia Alba na humane limfocite in vitro i SW-480 ćelijsku liniju karcinoma kolona
Investigation of the genotoxic and cytotoxic effects of Artemisia vulgaris and Artemisia alba spesies on human lymphocytes in vitro and SW-480 colon cancer cell line
Докторанд
Radović Jakovljević, MarinaМентор
Milošević-Đorđević, OliveraЧланови комисије
Joksić, GordanaGrujičić, Darko
Stanković, Milan
Метаподаци
Приказ свих података о дисертацијиСажетак
Artemisia vrste se tradicionalno koriste u tretmanu različitih bolesti. Cilj ove
doktorske disertacije bio je da se ispita fitohemijski sastav i genotoksični i
citotoksični efekat ekstrakata (metanolski, acetonski i vodeni) Artemisia vulgaris L.
i Artemisia alba Turra pojedinačno i u kombinovanom tretmanu sa mitomicinom C
(MMC). Fitohemijski sastav je utvrđen spektrofotometrijski i primenom HPLC
metode. Citotoksičnost je procenjena MTT testom na ćelijskoj liniji kancera kolona
(SW-480) i mezenhimalnim matičnim ćelijama periodoncijuma (PDLS), dok je tip
ćelijske smrti utvrđen primenom protočne citometrije. Citokinezis-blok
mikronukleus (MN) test u kultivisanim humanim limfocitima je korišćen za
procenu genotoksičnog i antimutagenog efekta. Ekstrakti su bili bogati ukupnim
fenolima i flavonoidima, dok su se kvantitativnom analizom izdvojili metanolski
ekstrakti obe biljke kao najbogatiji. Najjači citostatski efekat posredovan ranom
apoptozom postignut je nakon tretmana malignih ...ćelija acetonskim ekstraktom A. alba.
Obe biljke ostvarile su sinergističko dejstvo sa MMC indukujući snažan
citotoksični i proapoptotski efekat na maligne ćelije. Vodeni ekstrakt A. alba nije
delovao genotoksično na humane limfocite, dok su u kombinovanom tretmanu sa MMC
ekstrakti obe biljke ispoljili antimutageni efekat. Analizom efekta najzastupljenije
fenolne kiseline (3,5-dihidroksibenzoeve kiseline) i flavonoida (kvercetin-3-Oglukopiranozida) uočeno je da je fenolna kiselina bila genotoksična samo u najvišoj,
dok je flavonoid povećavao MN frekvencu u svim testiranim koncentracijama. MMCindukovana MN frekvenca se redukovala nakon tretmana fenolnom kiselinom, dok je
flavonoid ispoljio komutageni efekat. Dobijeni rezultati ukazuju da je korišćenje
vodenog ekstrakta A. alba u lekovite svrhe bezbedno.
Аrtemisia species are used in traditional medicine to treat various diseases. The aim of this
dissertation was to investigate the phytochemical composition and genotoxic and cytotoxic
effect of different extracts (methanolic, acetone, and aqueous) of Artemisia vulgaris L. and
Artemisia alba Turra, separately and in combination with mitomycin C (MMC). The
phytochemical composition was determined spectrophotometrically and using the HPLC
method. Cytotoxicity was assessed by the MTT assay on a colon cancer cell line (SW-480) and
stem cells derived from the human periodontal ligament (PDLSCs), while the type of cell death
was analyzed by flow cytometry. The cytokinesis-block micronucleus (MN) assay in peripheral
blood lymphocytes was used to evaluate the genotoxic and antimutagen effect. The extracts
were rich in total phenolic and flavonoid, while quantitative analysis revealed that the
methanolic extracts were the most abundant. The great cytostatic effect mediated by early
ap...optosis was achieved after treatment with acetone extract A. alba. Both plants achieved a
synergistic effect with MMC, inducing strong cytotoxic and proapoptotic effect on malignant
cells. Aqueous extract of A. alba was not genotoxic on human lymphocytes, whereaes in the
combined treatment with MMC, both plants showed antimutagenic effect. Analysis of the
effect of the most abundant phenolic acid (3,5- dihydroxybenzoic acid) and flavonoid
(quercetin-3-O-glucopyranosid) showed that phenolic acid showed a genotoxic effect in the
highest, whereas flavonoid increased MN frequency at all tested concentrations. MMC-induced
MN frequency was reduced after treatment with phenolic acid, whereas flavonoid exhibited
significant co-mutagenic effect. The results showed that the use of aqueous extract of A. alba
for medicinal purposes is safe.