TY - JOUR
T1 - The oxysterol 27-hydroxycholesterol increases β-amyloid and oxidative stress in retinal pigment epithelial cells
AU - Dasari, Bhanu
AU - Prasanthi, Jaya Rp
AU - Marwarha, Gurdeep
AU - Singh, Brij B.
AU - Ghribi, Othman
N1 - Funding Information:
Supported by a grant from NIEHS/NIH to OG (5R01ES014826) and BBS (5RO1DE017102, and 5P20RR017699). The authors thank Dr. Byron Grove and Sarah Rolling (Department of Anatomy and Cell Biology, School of Medicine & Health Sciences, University of North Dakota) for their technical assistance in confocal microscopy.
PY - 2010
Y1 - 2010
N2 - Background. Alzheimer's disease (AD) and age-related macular degeneration (AMD) share several pathological features including -amyloid (A) peptide accumulation, oxidative damage, and cell death. The causes of AD and AMD are not known but several studies suggest disturbances in cholesterol metabolism as a culprit of these diseases. We have recently shown that the cholesterol oxidation metabolite 27-hydroxycholesterol (27-OHC) causes AD-like pathology in human neuroblastoma SH-SY5Y cells and in organotypic hippocampal slices. However, the extent to which and the mechanisms by which 27-OHC may also cause pathological hallmarks related to AMD are ill-defined. In this study, the effects of 27-OHC on AMD-related pathology were determined in ARPE-19 cells. These cells have structural and functional properties relevant to retinal pigmented epithelial cells, a target in the course of AMD. Methods. ARPE-19 cells were treated with 0, 10 or 25 M 27-OHC for 24 hours. Levels of A peptide, mitochondrial and endoplasmic reticulum (ER) stress markers, Ca2+ homeostasis, glutathione depletion, reactive oxygen species (ROS) generation, inflammation and cell death were assessed using ELISA, Western blot, immunocytochemistry, and specific assays. Results. 27-OHC dose-dependently increased A peptide production, increased levels of ER stress specific markers caspase 12 and gadd153 (also called CHOP), reduced mitochondrial membrane potential, triggered Ca2+ dyshomeostasis, increased levels of the nuclear factor B (NFB) and heme-oxygenase 1 (HO-1), two proteins activated by oxidative stress. Additionally, 27-OHC caused glutathione depletion, ROS generation, inflammation and apoptotic-mediated cell death. Conclusions. The cholesterol metabolite 27-OHC is toxic to RPE cells. The deleterious effects of this oxysterol ranged from A accumulation to oxidative cell damage. Our results suggest that high levels of 27-OHC may represent a common pathogenic factor for both AMD and AD.
AB - Background. Alzheimer's disease (AD) and age-related macular degeneration (AMD) share several pathological features including -amyloid (A) peptide accumulation, oxidative damage, and cell death. The causes of AD and AMD are not known but several studies suggest disturbances in cholesterol metabolism as a culprit of these diseases. We have recently shown that the cholesterol oxidation metabolite 27-hydroxycholesterol (27-OHC) causes AD-like pathology in human neuroblastoma SH-SY5Y cells and in organotypic hippocampal slices. However, the extent to which and the mechanisms by which 27-OHC may also cause pathological hallmarks related to AMD are ill-defined. In this study, the effects of 27-OHC on AMD-related pathology were determined in ARPE-19 cells. These cells have structural and functional properties relevant to retinal pigmented epithelial cells, a target in the course of AMD. Methods. ARPE-19 cells were treated with 0, 10 or 25 M 27-OHC for 24 hours. Levels of A peptide, mitochondrial and endoplasmic reticulum (ER) stress markers, Ca2+ homeostasis, glutathione depletion, reactive oxygen species (ROS) generation, inflammation and cell death were assessed using ELISA, Western blot, immunocytochemistry, and specific assays. Results. 27-OHC dose-dependently increased A peptide production, increased levels of ER stress specific markers caspase 12 and gadd153 (also called CHOP), reduced mitochondrial membrane potential, triggered Ca2+ dyshomeostasis, increased levels of the nuclear factor B (NFB) and heme-oxygenase 1 (HO-1), two proteins activated by oxidative stress. Additionally, 27-OHC caused glutathione depletion, ROS generation, inflammation and apoptotic-mediated cell death. Conclusions. The cholesterol metabolite 27-OHC is toxic to RPE cells. The deleterious effects of this oxysterol ranged from A accumulation to oxidative cell damage. Our results suggest that high levels of 27-OHC may represent a common pathogenic factor for both AMD and AD.
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U2 - 10.1186/1471-2415-10-22
DO - 10.1186/1471-2415-10-22
M3 - Article
C2 - 20836858
AN - SCOPUS:77956494238
SN - 1471-2415
VL - 10
JO - BMC Ophthalmology
JF - BMC Ophthalmology
IS - 1
M1 - 22
ER -