Dual redox modulation by trolox and melatonin in glioma cells: a multi-parametric analysis with T98G validation

Oxidative stress, inflammation, and impaired apoptotic signaling jointly contribute to glioma progression and treatment resistance. This study compared the effects of Trolox and melatonin on redox balance, inflammatory signaling, and apoptosis in U87-MG glioma cells using a comprehensive multi-parameter phenotypic panel, evaluated their potential complementary actions, and validated key findings in T98G cells. U87-MG cells were treated with Trolox (25-200 µM), melatonin (25-200 µM), or their combination (100 µM Trolox + 100 µM melatonin) for 6, 24, and 48 h. Cell viability, oxidative stress indices (total oxidant status, total antioxidant capacity, oxidative stress index), intracellular ROS and lipid peroxidation markers (MDA, 4-HNE), inflammatory cytokines (IL-6, TNF-α), apoptosis, and expression of Nrf2-HO-1-SOD2 and Bax/Bcl-2 were analyzed. NF-κB p65 activity and caspase-3/7 activation were assessed to clarify inflammatory and apoptotic signaling, with N-acetylcysteine used as a reference thiol antioxidant in selected assays. Melatonin induced dose-dependent antioxidant, anti-inflammatory, and pro-apoptotic effects at lower concentrations than Trolox, whereas Trolox primarily suppressed lipid peroxidation at higher doses. The combination treatment consistently produced the most pronounced effects across endpoints, including maximal oxidative stress reduction, NF-κB inhibition, and apoptosis induction. Similar response patterns were confirmed in T98G cells, while no cytotoxicity was observed in normal human astrocytes. These findings support dual redox modulation as a mechanistically rational adjuvant strategy in glioma research.

The online version contains supplementary material available at 10.1007/s10616-026-00915-x.