Aqueous olive leaf extract (AOLE) of the Mediterranean tree Olea europaea is stated to have several therapeutic properties including: antifungal, antimicrobial, hypotensive and hypoglycemic actions as well as to use in treatment of some cancer cases as breast, skin and pancreatic cancers. Experimentally, the olive leaf extract is capable to protect nuclear calf thymus and human genomic DNA in vitro to Rhodamine B toxicity. Thus, this study was aimed to evaluate the protective effect of AOLE against toxic DNA damage induced by Rhodamine B using different techniques as UV−VIS Spectroscopy and fluorometry. Genomic DNA was used in a concentration of 75µg/ ml. DNA was extracted from peripheral lymphocytes of anticoagulated blood (EDTA) samples by Proteinase K digestion and phenol/chloroform extraction. Dried leaves of Olea europaea (from West part of Libya) were grinded with distilled water; the resulting aqueous solution was then filtered and used. The protective effect of the AOLE was evaluated for its ability to decrease the amount of Rhodamine B available for binding with double stranded DNA. Thus, DNA resulted in a significant decrease in fluorescence intensity of Rhodamine B. Comparing to control samples, pre-and post-treatment with AOLE, significantly attenuated the binding ability of Rhodamine B to the DNA (p<0.01). Treatment with 5g/15 ml of AOLE was enough to prevent Rhodamine B toxicity to the DNA. The cytoprotective and antioxidant properties were detected in AOLE by a fluorescence method. The antioxidant activity of AOLE was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and the IC 50 was 65±2.3μg/ml. In conclusion, this study suggests that AOLE induced a genoprotective effect by increasing the antioxidant capacity and the chemical interaction between Rhodamine B and constituents of AOLE.
Phospholipase A2 (PLA2) represents a major venom component of snakes and bees and exhibits a broad range of biological effects including myotoxicity, neurotoxicity, hemolysis, cardiotoxicity, anticoagulant and antiplatelet activities. Melittin, is the main component and the major pain producing substance of honeybee venom. The aim of the present study was to differentiate between snake and bee venoms using Aqueous Olive Leaf Extract (AOLE) employing fluorescence techniques. Tryptophan, which is contained in both snake and bee venoms is fluorescent at UV wavelength and hence widely used as a tool to monitor conformational changes in proteins and to draw inferences regarding local structure and dynamics. Fluorescence spectroscopy and molecular modeling have been used to analyze enzyme activity in the absence and presence of AOLE and to verify potential binding of AOLE components to the enzyme. Changes in the fluorescence intensities with blue and red shifts were obtained with bee and snake venoms, respectively. Binding of AOLE constituents near the active site of the enzyme could be evidenced and possible modes of interaction are discussed. The fluorescence method used was rapid and sensitive and was able to differentiate between snake and bee venoms utilizing AOLE.
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