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목적 : To investigate role of autophagy in the pathogenesis, especially the adipogenesis of orbital fibroblasts from Graves’ orbitopathy (GO) patients.
방법 : Autophagy related genes were identified and compared between both non-GO and GO orbital tissues using real time PCR, and electron microscopic examinations were performed to find autophagosoms in both non-GO and GO orbital fibroblasts. Cells were treated with bafilomycin A1, which inhibits autophagosome-lysosome fusion during adipogenesis to reduce autophagic flux. After induction of adipogenesis after pharmacologic manipulation of autophagy, adipogenesis related proteins and lipid droplet accumulation was identified by western blot analysis and oil red O stains.
결과 : LCII, p62 and Atg 5 mRNA levels were expressed significantly higher in GO tissues than in non-GO tissues. In electron microscopic examination, more autophagic vacuoles were frequently found in GO cells. When cells were treated with bafilomycin A1, LC3II and p62 proteins increased significantly, especially in GO cells. During differentiation, LCII and p62 proteins were all strongly produced in the early phase. After the flux inhibition, LC3II and p62 proteins increased consistently during adipogenesis, and both lipid droplet accumulation and adipogenesis markers decreased significantly.
결론 : Increased autophagy was observed in GO tissue and cells than in non-GO. Also, autophagy observed in GO orbital fibroblasts involved increased autophagosome formation and autophagy related proteins at early phase of differentiation and increased autophagic flux throuout 10 day of differentiation. Our study indicates that autophagy is involved in adipogenesis of GO orbital fibroblasts.
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