Zbornik - Društvo genetičara Srbije

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38 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE I-Pos-17 Subotica, 30. novembar - 4. decembar 2004. ANTIMUTAGENA SVOJSTVA PRIRODNIH ANTIOKSIDANATA DETEKTOVANA WP2 TESTOM ANTIMUTAGENEZE B. Vukoviæ-Gaèiæ, J. Stanojeviæ, J. Kneeviæ-Vukèeviæ, D. Mitiæ-Æulafiæ, T. Beriæ-Bjedov, B. Nikoliæ, S. Stankoviæ i D. Simiæ Katedra za mikrobiologiju, Biološki fakultet, Univerzitet u Beogradu, Beograd Oksidativna ošteæenja DNK, izazvana reaktivnim kiseoniènim vrstama (ROS) su veoma znaèajna u procesima mutageneze, kancerogeneze i starenja. Ispitivanja aktivnih supstanci lekovitih i zaèinskih biljaka su potvrdila da mnoge poseduju antioksidativnu aktivnost i da su potentni inhibitori mutageneze i kancerogeneze. U ovom radu korišæen je WP2 test antimutageneze (WP2 Antimutagenicity test) za praæenje oksidativne mutageneze i kvantifikovanje antimutagenog potencijala antioksidanata. Ispitivanje je sprovedeno na dva soja E.coli: IC185 (WP2 oxyR + )i njegovom derivatu IC202 (WP2 oxyR/pKM101). Oksidativna mutageneza je indukovana tert-butil hidroperoksidom (t-BOOH), latentnim donorom RO' alkoksil radikala. Zbog defekta u OxyR funkciji, soj IC202 je mnogo osetljiviji na oksidativna ošteæenja, te je pokazao veæu mutabilnost u poreðenju sa OxyR + sojem. Vitamin E (-tokoferol) je korišæen kao model antioksidant jer je veoma efikasan sakupljaè (‘scavenger’) ROS-a i on je testiran u oba soja. Nakon validacije test sistema vitaminom E, korišæen je samo osetljiviji oxyR - soj za ispitivanje antimutageneze posredovane antioksidativnim svojstvima dva derivata bosiljka (Ocimum basilicum L.): etarskog ulja (EO) i njegovog najzastupljenijeg monoterpena linaloola. Vitamin E je inhibirao indukovanu mutagenezu u oba soja (63% u soju IC202 i 43% u soju IC185). Oba derivata bosiljka su pokazala snaan antimutageni potencijal prema indukovanoj oksidativnoj mutagenezi (55% inhibicije je izazvalo EO, a 68% linalool). ANTIMUTAGENIC PROPERTIES OF NATURAL ANTIOXIDANTS IN THE WP2 ANTIMUTAGENICITY TEST Oxidative damage to DNA by reactive oxygen species (ROS) is very important in mutagenic, carcinogenic, and aging processes. The study of active substances from medicinal and aromatic plants has revealed that many of them possess antioxidative activity and may be potent inhibitors of mutagenesis and carcinogenesis. In this work we used WP2 antimutagenicity test for detection of oxidative mutagenesis and antimutagenic potential of antioxidants. Screening was performed using E.coli IC185 (WP2 oxyR + ) and its counterpart IC202 (WP2 oxyR/pKM101). Oxidative mutagenesis was induced by t-butyl hydroperoxide (t-BOOH), the latent donor of RO' alkoxyl radicals. Strain IC202, highly sensitive to oxidative damage due to its deficiency in the OxyR function, shows increased mutability with respect to the OxyR+ strain. Vitamin E (a-tocopherol), potent scavenger of reactive oxygen species, was used as the model antioxidant and it was tested in both tester strains. After validation, we used highly sensitive oxyR - strain to test essential oil (EO) of basil (Ocimum basilicum L.) and its major monoterpene – linalool for antimutagenesis due to their antioxidative properties. Vitamin E inhibited induced mutagenesis in both strains (63% in IC202 and 43% in IC185). Tested basil derivatives also exhibited strong antimutagenic potential against induced oxidative mutagenesis (55% of inhibition for EO and 68% for linalool).
I-Pos-18 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 39 Subotica, 30. novembar - 4. decembar 2004. DETEKCIJA ANTIOKSIDATIVNE AKTIVNOSTI SUPSTANCI PRIRODNOG POREKLA NOVIM TEST-SISTEMOM Jasna Stanojeviæ 1 , Jelena Kneeviæ-Vukèeviæ 1 i G. Miloshev 2 1 Katedra za mikrobiologiju, Biološki fakultet, Univerzitet u Beogradu, Beograd 2 Institut za molekularnu biologiju, Bugarska Akademija Nauka, Sofija, Bugarska Reaktivne kiseoniène vrste endogenog i egzogenog porekla mogu biti ukljuèene u proces starenja i nastanak razlièitih humanih bolesti kao što su ateroskleroza, kancer, dijabetes, neurodegenerativna oboljenja i sindrom steèene imunodeficijencije. Antioksidanti koji se nalaze u lekovitim i aromatiènim biljkama, voæu i povræu mogu imati blagotvoran efekat na ljudsko zdravlje i mogu biti korisni za prevenciju štetnih posledica oksidativnih ošteæenja izazvanih reaktivnim kiseoniènim vrstama. U ovom radu je korišæen Comet test na kvascu (YCA), metoda razvijena od strane Miloshev et al. [1], za detekciju antioksidanata sa inhibitornim efektom na nastanak oksidativnih ošteæenja indukovanih vodonik peroksidom. Vitamin E (-tokoferol) i vitamin C (askorbinska kiselina) su korišæeni kao model antioksidanti. Nakon validacije, testiran je linalol glavni konstituent etarskog ulja bosiljka (Ocimum basilicum L). Sposobnost antioksidanata da smanje nivo oksidativnih ošteæenja DNK ispitivana je primenom pre-tretmana i ko-tretmana (istovremeno dodavanje antioksidanta i oksidanta). Vitamin E koji je znaèajno snizio broj indukovanih oksidativnih ošteæenja, jaèi inhibitorni efekat je pokazao u pre-tretmanu (81%) u poreðenju sa ko-tretmanom (43%). Snana sposobnost inhibicije H2O2-indukovanih DNK ošteæenja je detektovana i za vitamin C, ali procenat inhibicije dostie istu vrednost (65%) i za pre-tretman i za ko-tretman. Monoterpen linalol je takoðe pokazao jaku antioksidativnu aktivnost. Detektovan inhibitorni efekat se kreæe u opsegu od 55% za ko-tretman do 70% za pre-tretman. Na osnovu dobijenih rezultata linalol se moe svrstati meðu supstance sa veoma snanim antioksidativnim dejstvom èiji bi potencijalni antikancerogeni efekat trebalo dalje ispitati. DETECTION OF ANTIOXIDATIVE ACTIVITIES OF NATURAL SUBSTANCES WITH A NEWLY DEVELOPED SYSTEM Reactive oxygen species (ROS), from both endogenous and exogenous sources, may be involved in aging and different human diseases such as atherosclerosis, cancer, diabetes, neurodegenerative diseases and acquired immunodeficiency syndrome (AIDS). Natural antioxidants contained in medicinal and aromatic plants, fruits and vegetables may have beneficial health effect and may be useful in preventing the deleterious consequences of oxidative damage caused by ROS. In this work Yeast Comet Assay (YCA), a method developed by Miloshev et al. [1], is used for the detection of antioxidative activity of different compounds against hydrogen peroxide-induced oxidative damage. Vitamin E (-tocopherol) and vitamin C (ascorbic acid) were used as model antioxidants. After validation, we have tested linalool major constituent of basil (Ocimum basilicum L). The ability of antioxidant pre-treatment and co-treatment (simultaneously with oxidant) to decrease DNA damage was assessed. Vitamin E exhibited the protective effect against induced oxidative damage; the stronger inhibitory effect was detected in pre-treatment (81%) comparing with co-treatment (43%). Vitamin C also displayed a significant protective capability against H2O2-induced DNA damage. However percentage of inhibition reached the same value (65%) both for co-treatment and pre-treatment. Linalool, the oxygen-containing monoterpene, showed also very strong antioxidative activity. Inhibitory effect is in the range from 55% for co-treatment to 70% for pre-treatment. These results put linalool as one of substances with very strong antioxidative acting and recommend it for further anticarcinogenic studies.
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Zbornik - Društvo genetičara Srbije

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