Cytoprotective effects and antioxidant activities of acteoside and various extracts of Clerodendrum cyrtophyllum Turcz leaves against t-BHP induced oxidative damage

This study evaluates the antioxidant potential and cytoprotective effects of ethanolic crude extract from Clerodendrum cyrtophyllum leaves (ECE) and five derived fractions (namely, petroleum ether fraction (PEF), dichloromethane fraction (DMF), ethyl acetate fraction (EAF), n-butyl alcohol fraction (BAF) and the remaining fraction (RF)), as well as acteoside (Ac, a major phenolic component in EAF) on oxidative damage caused by tert-butyl hydroperoxide (t-BHP) in HepG2 cells. MTT assay results showed that ECE, EAF, BAF, RF and Ac increased the viability of t-BHP-damaged cells in a dose-dependent manner, while EAF significantly promoted cell viability. EAF, BAF, RF, or Ac reduced the levels of lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA), and reactive oxygen species (ROS). Additionally, glutathione (GSH) levels and the activities of superoxide dismutase (SOD) and catalase (CAT) increased. Western blot analysis further indicated that EAF, BAF, RF, or Ac up-regulated pro-caspase-3 and reduced cleaved caspase-3 during t-BHP-induced oxidative stress. Flow cytometry analysis and fluorescence micrographs showed that Ac could inhibit apoptosis.