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Therapeutic effect of protocatechuic aldehyde on graves' orbitopathy in an in vitro model

Authors
 변정우 
Issue Date
2015
Description
Dept. of Medical Science/석사
Abstract
Graves’ disease (GD) is an autoimmune disorder caused by excessive secretion of thyroid hormones. Pathological symptoms are observed in the eyes of the patients with GD, and these symptoms are called “Graves’ orbitopathy” (GO). The general symptom of GO is exophthalmos caused by increased connective tissue volume within the orbital. I investigated the therapeutic effect of protocatechuic aldehyde (3,4-dihydroxybenzaldehyde; PCA ) in an in vitro GO model. The experiments were carried out using orbital fibroblasts, which were cultured primarily from orbital fat tissue obtained after orbital decompression.I performed a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to confirm the anti-oxidizing effect of PCA, and an 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymeth`oxyphe nyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay to confirm the effect of PCA on the viability of normal and GO orbital fibroblasts. The anti-oxidant effect of PCA on reactive oxygen species (ROS) generation was stimulated with hydrogen peroxide (H2O2) and detected with 5-(and-6)-carboxy-2'',7''-difluorodihydrofluorescein diacetate (carboxy-H2DFFDA) by fluorescence activated cell sorting analysis. In addition, the expression of anti-oxidant enzyme heme oxygenase (HO) -1 was confirmed. The inhibitory effect of PCA on tumor necrosis factor (TNF) -α induced intercellular adhesion molecule (ICAM) -1 expression and hyaluronan production was determined by western blot analysis and a hyaluronan enzyme-linked immunosorbent assay (ELISA), respectively. To evaluate anti-adipogenic activities, PCA was added to adipose differentiation media, which included peroxisome proliferator activator gamma (PPAR-γ) agonist. After differentiation, the cells were stained with Oil red O, and adipose differentiation related protein expression of PPAR-γ, CCAAT-enhancerbinding proteins (c/EBP) -α, and -β,

was confirmed by western blot analysis. DPPH assay results confirmed the free radical scavenging effect of PCA after treatment. Results of the MTS assay confirmed that PCA did not have any significant effect on cell viability. However, PCA had a suppressive effect on intracellular ROS generation and upregulated HO-1 expression by western blot analysis. PCA attenuated TNF-α induced ICAM-1 protein expression and inhibited hyaluronan production in a dose-dependent manner. Based on the results of Oil Red O staining, treatment with PCA during adipose differentiation, caused an decrease in lipid droplets, and expression of related proteins, including PPAR-γ c/EBP-α, and c/EBP-β was suppressed. I confirmed that PCA has anti-inflammatory, anti-oxidant, anti-adipogenic and inhibitory effects on hyaluronan production in vitro. With further research and clinical studies, I believe that PCA can potentially be used as a novel therapy for GO.
Full Text
https://ymlib.yonsei.ac.kr/catalog/search/book-detail/?cid=CAT000000226310
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Appears in Collections:
1. College of Medicine (의과대학) > Others (기타) > 2. Thesis
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/146106
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