An Optimum Dose of Olive Leaf Extract Improves Insulin Receptor Substrate-1, Tyrosine Kinase, and Glucose Transporters, While High Doses Have Genotoxic and Apoptotic Effects — Oak Academic Publishing
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An Optimum Dose of Olive Leaf Extract Improves Insulin Receptor Substrate-1, Tyrosine Kinase, and Glucose Transporters, While High Doses Have Genotoxic and Apoptotic Effects
Department of Medical Biochemistry, Medical Faculty, Bezmialem Vakif University, Istanbul, Turkey
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Department of Medical Biochemistry, Medical Faculty, Bezmialem Vakif University, Istanbul, Turkey
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Traditional and Complementary Medicine Advanced Research and Application Center (GETAMER), Bezmialem Vakif University, Istanbul, Turkey
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Department of Molecular Biology, Genetics, and Bioengineering, Sabanci University, Istanbul, Turkey
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Republic of Turkey Ministry of Agriculture and Forestry Istanbul Pendik Veterinary Control Institute, Istanbul, Turkey
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Department of Plant Protection, Agricultural Faculty, Harran University, S. Urfa, Turkey
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Department of Molecular Biology and Genetics, Gebze Technical University, Kocaeli, Turkey
1 Department of Medical Biochemistry, Medical Faculty, Bezmialem Vakif University, Istanbul, Turkey
2 Department of Medical Biochemistry, Medical Faculty, Bezmialem Vakif University, Istanbul, Turkey
3 Traditional and Complementary Medicine Advanced Research and Application Center (GETAMER), Bezmialem Vakif University, Istanbul, Turkey
4 Department of Molecular Biology, Genetics, and Bioengineering, Sabanci University, Istanbul, Turkey
5 Republic of Turkey Ministry of Agriculture and Forestry Istanbul Pendik Veterinary Control Institute, Istanbul, Turkey
6 Department of Plant Protection, Agricultural Faculty, Harran University, S. Urfa, Turkey
7 Department of Molecular Biology and Genetics, Gebze Technical University, Kocaeli, Turkey
Type 2 diabetes is the most common type of diabetes. Conventionally many drugs are used for the treatment of diabetes such as biguanides, sulfonylureas, meglitinides, etc. But the desired effective treatment is still not to be achieved. So researches are going on for the development of effective alternative therapy against diabetes. Olive leaves are traditionally used in the treatment of the disease. However, studies on its mechanism of action are not yet enough. The aim of this study was to investigate whether olive leaf extract (OLE) improves insulin receptor substrate-1 (IRS-1), tyrosine kinase (TK), GLUT-2, and GLUT-4. Oleuropein levels were analyzed from OLE obtained by using four different solvents, and the highest content of methanol extract was selected for the study. Different concentrations of OLE (2.5 to 320 μ g/mL) were incubated with hepatocellular carcinoma (HepG2) cells for 24 hours. After incubation, cell viability was assessed based on luminometric ATP cell viability assay kit. Intracellular reactive oxygen species (ROS) generating level was detected using 2,7dichlorodihydrofluorescein-diacetate (H 2 DCF-DA) fluorescent probes. Apoptosis was evaluated by acridine orange/ethidium bromide double staining method. Genotoxicity was evaluated by alkaline single cell gel electrophoresis assay (Comet Assay). Protein expression levels of IRS-1, TK, GLUT-2, and GLUT-4 were analyzed by western blotting technique from the obtained cell lysates. Although an optimum doses of OLE (10 μ g/mL) maximally increased cell proliferation, decreased ROS generation improved IRS-1, TK, GLUT-2, and GLUT-4 protein expression levels (about fivefold), higher doses (10 to 320 μ g/mL) markedly decreased the cell viability, increased DNA damage, apoptosis and ROS generation in a concentration-dependent manner. OLE can be used in the treatment of type 2 diabetes. However, in order to find the most effective and non-toxic concentration, dose optimization is required.
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