Protective effect of a novel cystine C(60) derivative on hydrogen peroxide-induced apoptosis in rat pheochromocytoma PC12 cells.
Oxidative stress has been considered as a major cause of cellular injuries in a variety of clinical abnormalities, especially neural diseases. One of the effective ways to prevent the reactive oxygen species (ROS) mediated cellular injury is dietary or pharmaceutical augmentation of free radical scavengers. In the present study, we describe the synthesis and characterization of a novel cystine C(60) derivative (CFD). The compound was analyzed by FT-IR, (1)H NMR, (13)C NMR, LC-MS and elemental analysis. It contains five cystine moieties per C(60) molecule. This water-soluble amino-fullerene derivative was able to scavenge both superoxide and hydroxyl radical with biocompatibility. We investigated its potential protective effects on hydrogen peroxide-induced oxidative stress and apoptotic death in cultured rat pheochromocytoma (PC12) cells. Cells treated with hydrogen peroxide underwent cytotoxicity and apoptotic death determined by MTT assay, flow cytometry analysis, PI/Hoechst 33342 staining and glutathione peroxidase assay. The CFD was able to reduce the accumulation of reactive oxygen species and cellular damage caused by hydrogen peroxide in PC12 cells. RF assay demonstrated that CFD could penetrate through the cell membrane and it has played its distinguished role in protecting PC12 cells against hydrogen peroxide-induced cytotoxicity. The results suggest that CFD has the potential to prevent oxidative stress-induced cell death without evident toxicity. Hence, we can hypothesize that the protective effect of CFD on hydrogen peroxide-induced apoptosis is related to its scavenger activity.
Hu
Zhen
Z
Department of Chemistry, Huazhong University of Science and Technology, Wuhan 430074, PR China.
Guan
Wenchao
W
Wang
Wei
W
Huang
Lizhen
L
Xing
Haiping
H
Zhu
Zhou
Z
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D003553:Cystine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D005562:Formazans / Q000737:chemistry; D016166:Free Radical Scavengers / Q000737:chemistry / Q000494:pharmacology; D037741:Fullerenes / Q000737:chemistry / Q000494:pharmacology*; D005979:Glutathione Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D018384:Oxidative Stress / Q000187:drug effects*; D016716:PC12 Cells; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013778:Tetrazolium Salts / Q000737:chemistry
D005456:Fluorescent Dyes; D005562:Formazans; D016166:Free Radical Scavengers; D037741:Fullerenes; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; C079782:MTT formazan; D003553:Cystine; D006861:Hydrogen Peroxide; D005979:Glutathione Peroxidase; C069837:fullerene C60