Cholesterol secoaldehyde, an ozonation product of cholesterol, induces amyloid aggregation and apoptosis in murine GT1-7 hypothalamic neurons.
Aldehydic products from ozonation of cholesterol and peroxidation of phospholipids have been shown to accelerate aggregation of amyloid-beta (Abeta) in vitro. Here, we show that 3beta-hydroxy-5-oxo-5,6-secocholestan-6-al (ChSeco), an ozonation product of cholesterol, induces Abeta aggregation, generation of reactive oxygen species (ROS), and cytotoxicity in murine GT1-7 hypothalamic neurons. The formation of Abeta aggregates in situ was dose-dependent at ChSeco concentrations ranging from 1 to 20 microM. The increase in insoluble Abeta aggregates at increasing concentrations of ChSeco was accompanied by a decrease in soluble Abeta as evidenced by Western blot analysis. The formation of ROS in neuronal cells was found to be dose- and time-dependent with the magnitude being higher at 20 microM compared to 10 microM ChSeco or untreated controls. The increase in ROS was associated with depletion of GSH. The cytotoxicity induced by ChSeco involved changes in phosphatidylserine translocation, DNA fragmentation, and caspase 3/7 activity that are characteristic of apoptosis. Pretreatment of neuronal cells with Trolox, a water-soluble analog of alpha-tocopherol offered partial, but significant protection against ChSeco-induced cell death, whereas, N-acetyl-L-cysteine (NAC) completely prevented the cytotoxic effects of ChSeco. NAC and Trolox were without any effects on ChSeco-induced Abeta aggregation. Fibrillogenesis inhibitors, which inhibited Abeta aggregation, did not inhibit cell death induced by ChSeco, implying that ROS generation, and not Abeta aggregation, plays a major role in the observed cytotoxicity. However, since Alzheimer's and other neurodegenerative diseases are slow and progressive, the formation of Abeta aggregates in vivo by ChSeco may have long-term pathological consequences.
Sathishkumar
K
K
Department of Environmental Toxicology and The Health Research Center, Southern University and A&M College, Baton Rouge, Louisiana 70813, USA.
Xi
Xiaochun
X
Martin
Roy
R
Uppu
Rao M
RM
D000111:Acetylcysteine / Q000008:administration & dosage / Q000494:pharmacology; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002449:Cell Aggregation / Q000502:physiology*; D002778:Cholestanones / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D008076:Cholesterol, HDL / Q000187:drug effects / Q000378:metabolism; D002839:Chromans / Q000008:administration & dosage / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D016166:Free Radical Scavengers / Q000008:administration & dosage / Q000494:pharmacology; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D015227:Lipid Peroxidation; D051379:Mice; D032386:Myoblasts, Cardiac / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D009474:Neurons / Q000378:metabolism* / Q000473:pathology*; D010126:Ozone / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012632:Secosteroids / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019680:Trinucleotide Repeat Expansion / Q000502:physiology*
D016229:Amyloid beta-Peptides; D000975:Antioxidants; D002778:Cholestanones; D008076:Cholesterol, HDL; D002839:Chromans; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D012632:Secosteroids; D010126:Ozone; C082684:3-hydroxy-5-oxo-5,6-secocholestan-6-al; D002784:Cholesterol; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D000111:Acetylcysteine