Fullerenol Protects Human Retinal Pigment Epithelial Cells from Oxidative-Stress-Induced Premature Senescence Model in vitro

Abstract

Purpose: : Chemical modification of fullerenes to hydrosoluble cluster molecules made fullerenes interesting for biological applications Fullerenol was powerful antioxidant: it reacted with superoxide anion radical, hydroxy radical and nitrous oxide radical in compounds and epithelium (RPE) is one of the major suffered cells in human age-related macular degeneration (AMD). Taken in consideration together, fullerenol is a promising candidate in AMD therapy. This study is aimed to determine whether fullerenol can protect (RPE) cells from oxidative-stress-induced death.

Methods: : The cytotoxity, efficacy and potency were determined of fullerenol on the survival of human ARPE-19 cells treated with either hydrogen peroxide (H2O2) or t-butyl hydroperox-ide (t-BOOH).MTT assay was performed to evaluate the cell viabilities of the H2O2 and t-BOOH after 24 hours, Sublethal dose were used for establishment the in vitro model in our study. After exposure to 100uM H2O2 for 2h daily for 5days or 800 uM tert-butylhydroperoxide (tert-BHP) for 1 h daily for 5 days, four well-known senescence biomarkers were analysed: hypertrophy, senescence-associated β-galactosidase activity, growth arrest, and cell cycle arrest in G2/M. The senescence related genes and proteins were determined by real-time PCR and western blot.

Results: : With 5ug/ml fullerenol treatment, the sencescene biomarkers in the model can be alleviated. β-galactosidase activity and staining in fullerenol group is weaker than in model group. FACs showed fullerenol treatment break the G2/M arrest in model . Fullerenol treatment can rescues the oxidative-stress-induced sencescene genes and proteins changes, such as PCNA and CDC 2 which were suppressed by ROS

Conclusions: : 5 ug/mL fullerenol in culture medium don't show any toxicity to cells. Fullerenol can break the G2/M arrest though down regulating the sencescene related cell-cycle genes. The results demonstrated fullerenol can protected RPE cell from oxidative-stress induced sencescene with high degree of potency and low toxicity. Since Fullerenol is a nano-sized, long retention with low toxicity, it may be a candidate for AMD treatment.

Keywords: retinal pigment epithelium • aging • oxidation/oxidative or free radical damage