Protective effect of reduced glutathione C₆₀ derivative against hydrogen peroxide-induced apoptosis in HEK 293T cells

Abstract

Hydrogen peroxide (H₂O₂) and free radicals cause oxidative stress, which induces cellular injuries, metabolic dysfunction, and even cell death in various clinical abnormalities. Fullerene (C₆₀) 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 (C₆₀-GSH) was successfully synthesized, and its beneficial roles in protecting against H₂O₂-induced oxidative stress and apoptosis in cultured HEK 293T cells were investigated. Fourier Transform infrared spectroscopy and ¹H nuclear magnetic resonance were used to confirm the chemical structure of C₆₀-GSH. Our results demonstrated that C₆₀-GSH prevented the reactive oxygen species (ROS)-mediated cell damage. Additionally, C₆₀-GSH pretreatment significantly attenuated H₂O₂-induced superoxide dismutase (SOD) consumption and malondialdehyde (MDA) elevation. Furthermore, C₆₀-GSH inhibited intracellular calcium mobilization, and subsequent cell apoptosis via bcl-2/bax-caspase-3 signaling pathway induced by H₂O₂ stimulation in HEK 293T cells. Importantly, these protective effects of C₆₀-GSH were superior to those of GSH. In conclusion, these results suggested that C₆₀-GSH has potential to protect against H₂O₂-induced cell apoptosis by scavenging free radicals and maintaining intracellular calcium homeostasis without evident toxicity.

Keywords

reduced glutathione C₆₀ derivative / hydrogen peroxide / oxidative stress / apoptosis / HEK 293T cells

References

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