Cell-Type-Specific Modulation of Hydrogen Peroxide Cytotoxicity and 4-Hydroxynonenal Binding to Human Cellular Proteins In Vitro by Antioxidant Extract.

Although contains numerous bioactive components, the activity principles of widely used extracts are uncertain. Therefore, we analyzed the effects of genuine aqueous extract (AV) on human cells with respect to the effects of hydrogen peroxide (H₂O₂) and 4-hydroxynonenal (HNE). Fully developed leaves were harvested and analyzed for vitamin C, carotenoids, total soluble phenolic content, and antioxidant capacity. Furthermore, human cervical cancer (HeLa), human microvascular endothelial cells (HMEC), human keratinocytes (HaCat), and human osteosarcoma (HOS) cell cultures were treated with AV extract for one hour after treatment with H₂O₂ or HNE. The cell number and viability were determined using Trypan Blue, and endogenous reactive oxygen species (ROS) production was determined by fluorescence, while intracellular HNE⁻protein adducts were measured for the first time ever by genuine cell-based HNE⁻His ELISA. The AV extract expressed strong antioxidant capacities (1.1 mmol of Trolox eq/g fresh weight) and cell-type-specific influence on the cytotoxicity of H₂O₂, as well as on endogenous production of ROS and HNE⁻protein adducts induced by HNE treatment, while AV itself did not induce production of ROS or HNE⁻protein adducts at all. This study, for the first time, revealed the importance of HNE for the activity principles of AV. Since HMEC cells were the most sensitive to AV, the effects of AV on microvascular endothelia could be of particular importance for the activity principles of extracts.