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Effects of Oleate and Inflammatory Cytokines on Dermal Fibroblasts in Type 1 Diabetics
Author(s) -
Jones Albert Richard,
Deeney Jude T,
Corkey Barbara
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1010.9
Subject(s) - chemistry , lipid droplet , reactive oxygen species , inflammation , oil red o , peripheral blood mononuclear cell , stimulation , lipid oxidation , staining , medicine , biochemistry , endocrinology , adipose tissue , biology , pathology , in vitro , adipogenesis , antioxidant
Background Dermal fibroblasts are crucial to wound healing which can be impaired in type 1 diabetics. We set out to determine if exposure to inflammatory cytokines and oleate changed the metabolism and ROS production of fibroblasts. Methods Cells were acutely exposed to 25 or 50 μM of oleate and chronically exposed to 100 μM of oleate. Cells were exposed to 10 ng/mL of TNF‐ α. Oxygen Consumption was measured using the SeaHorse XF24 Flux Analyzer, ROS was measured using DCF dye. Lipid accumulation was measured using Nile Red staining. β‐oxidation was measured using radioactive oleate. Results The data shows type 1 diabetics sequester more lipid droplets than controls. As a result, less fatty acid is available for β‐oxidation and the stimulation of respiration, which explains the decreases in oxygen consumption, proton leak, β‐oxidation and ROS generation in diabetic fibroblasts. We believe that diabetics express an inflammatory phenotype that reduces β‐oxidation in the cell causing a decrease in oxygen consumption and ROS production. Future Research We plan to measure the mRNA and protein expression of key components of β‐oxidation and inflammation in fibroblasts. Funded by NIH