Protective effect of reduced glutathione C 60 derivative against hydrogen peroxide-induced apoptosis in HEK 293T cells

Summary Hydrogen peroxide (H 2 O 2 ) and free radicals cause oxidative stress, which induces cellular injuries, metabolic dysfunction, and even cell death in various clinical abnormalities. Fullerene (C 60 ) is critical for scavenging oxygen free radicals originated from cell metabolism, and reduced glutathione (GSH) is another important endogenous antioxidant. In this study, a novel water-soluble reduced glutathione fullerene derivative (C 60 -GSH) was successfully synthesized, and its beneficial roles in protecting against H 2 O 2 -induced oxidative stress and apoptosis in cultured HEK 293T cells were investigated. Fourier Transform infrared spectroscopy and 1 H nuclear magnetic resonance were used to confirm the chemical structure of C 60 -GSH. Our results demonstrated that C 60 -GSH prevented the reactive oxygen species (ROS)-mediated cell damage. Additionally, C 60 -GSH pretreatment significantly attenuated H 2 O 2 -induced superoxide dismutase (SOD) consumption and malondialdehyde (MDA) elevation. Furthermore, C 60 -GSH inhibited intracellular calcium mobilization, and subsequent cell apoptosis via bcl-2/bax-caspase-3 signaling pathway induced by H 2 O 2 stimulation in HEK 293T cells. Importantly, these protective effects of C 60 -GSH were superior to those of GSH. In conclusion, these results suggested that C 60 -GSH has potential to protect against H 2 O 2 -induced cell apoptosis by scavenging free radicals and maintaining intracellular calcium homeostasis without evident toxicity.