DNA PROTECTIVE ACTIVITY OF Artemisia spp HAIRY ROOT EXTRACTS

Abstract

Artemisia spp. plants are known as producers of bioactive compounds and used both in folk and traditional medicine. They possess antitumor, antiproliferative, antidiabetic, antimicrobial, antiviral, antioxidant, and anti-inflammatory activity. Aim. Artemisia spp. plants exhibit antioxidant activity, so it is of interest to investigate the possibility of using extracts from mugwort to prevent DNA damage initiated by some reactive oxygen species. Methods. In this work, extracts from transformed roots of A. vulgaris and A. tilesii were used to study their DNA protective activity. The extracts were prepared according to standard procedure. Total flavonoid content was quantified by the modified spectrophotometric method in rutin equivalent using the calibration curve. The antioxidant activity of the extracts was determined using 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). It was evaluated by the half maximal effective concentration (EC50) calculated as the dry root weight needed for scavenging 50% of DPPH in the sample and expressed as mg DW. To calculate this value, linear regression was applied to the linear interval of radical scavenging activity. DNA protective activity was studied by the Fenton reaction assay. Results. Differences in the content of flavonoids in A. vulgaris “hairy” roots and control roots were found. For hairy roots this parameter ranged from 75.89 ± 2.32 to 126.04 ± 5.37 mg RE/g DW, which is 1.45 - 2.41 times more than in the control roots. Flavonoid content in A. tilesii hairy root line also differed from the control. It was 74.52 ± 0.96 … 107.8 ± 5.98 mg RE/g DW in root lines and 28,6 ± 2,11 mg/g DW in A. tilesii control roots. The level of antioxidant activity studied in the reaction with DPPH (EC50, effective concentration) was more significant in the extracts of hairy roots of both plant species. It varied from 50 0.16-0.33 and 0.17-0.31 in hairy root lines of A. vulgaris and A. tilesii, respectively. In comparison, this parameter reached 0.44 and 0.65 in the control roots. Adding the extracts to the reaction mixture in the Fenton reaction has some protective effects. At the same time, there were no significant differences in the degree of protection of plasmid DNA from damage (percentage of supercoiled DNA) when extracts from hairy root lines of A. vulgaris and A. tilesii were added to the reaction mixture. However, these extracts differed in the content of flavonoids and had a higher ability to scavenge DPPH radicals. Conclusions. The extracts of A. vulgaris and A. tilesii hairy roots contained a higher concentration of flavonoids and had higher antioxidant activity compared to the extracts from the control roots. However, they differed little in their ability to protect DNA from damage in the Fenton reaction. Likely, that not only flavonoids, but also other components of extracts from wormwood hairy roots are involved in this process.