Knockdown of FABP5 mRNA decreases cellular cholesterol levels and results in decreased apoB100 secretion and triglyceride accumulation in ARPE-19 cells.
To maintain normal retinal function, retinal pigment epithelial (RPE) cells engulf photoreceptor outer segments (ROS) enriched in free fatty acids (FFAs). We have previously demonstrated fatty acid-binding protein 5 (FABP5) downregulation in the RPE/choroidal complex in a mouse model of aging and early age-related macular degeneration. FABPs are involved in intracellular transport of FFAs and their targeting to specific metabolic pathways. To elucidate the role of FABP5 in lipid metabolism, the production of the FABP5 protein in a human RPE cell line was inhibited using RNA interference technology. As a result, the levels of cholesterol and cholesterol ester were decreased by about 40%, whereas FFAs and triglycerides were increased by 18 and 67% after siRNA treatment, respectively. Some species of phospholipids were decreased in siRNA-treated cells. Cellular lipid droplets were evident and apoB secretion was decreased by 76% in these cells. Additionally, we discovered that ARPE-19 cells could synthesize and secrete Apolipoprotein B100 (apoB100), which may serve as a backbone structure for the formation of lipoprotein particles in these cells. Our results indicate that FABP5 mRNA knockdown results in the accumulation of cellular triglycerides, decreased cholesterol levels, and reduced secretion of apoB100 protein and lipoprotein-like particles. These observations indicated that FABP5 plays a critical role in lipid metabolism in RPE cells, suggesting that FABP5 downregulation in the RPE/choroid complex in vivo might contribute to aging and early age-related macular degeneration.
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. tingwu@jhmi.edu
Tinghuai
T
Wu
Jane
J
Tian
Roy G
RG
Cutler
Richard S
RS
Telljohann
David A
DA
Bernlohr
Mark P
MP
Mattson
James T
JT
Handa
D000818:Animals; D053299:Apolipoprotein B-100 / Q000378:metabolism; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism; D017931:DNA Primers; D050556:Fatty Acid-Binding Proteins / Q000172:deficiency* / Q000235:genetics*; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism; D008268:Macular Degeneration / Q000235:genetics / Q000628:therapy; D051379:Mice; D010743:Phospholipids / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics*; D034741:RNA, Small Interfering / Q000627:therapeutic use; D055213:Retinal Pigment Epithelium / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014280:Triglycerides / Q000378:metabolism*
D053299:Apolipoprotein B-100; D002788:Cholesterol Esters; D017931:DNA Primers; C493412:FABP5 protein, human; D050556:Fatty Acid-Binding Proteins; D005230:Fatty Acids, Nonesterified; D008074:Lipoproteins; D010743:Phospholipids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014280:Triglycerides; D002784:Cholesterol