S-Space College of Medicine/School of Medicine (의과대학/대학원) Dept. of Medicine (의학과) Theses (Ph.D. / Sc.D._의학과)
The Inhibitory Effect of Oligomeric Procyanidins(OPCs) on Procollagen Type I Secretion of Fibroblasts – In Vitro Study
- 의과대학 의학과
- Issue Date
- 서울대학교 대학원
- 학위논문 (박사)-- 서울대학교 대학원 : 의학과 성형외과학, 2013. 8. 김석화.
- Purpose: Wound healing is a very complex process that requires harmonies of various cell populations. It consists of complex series of events including inflammatory, proliferative, and remodeling phases. In final remodeling phase, fibroblasts play an important role of returning the wound to pre-injury state through collagen degradation. Oligomeric procyanidins (OPC) are main components of grape (Vitis vinifera) seed extracts and also found in wine and green tea. Recent studies showed OPCs effects on inflammation, cell migration and proliferation. From the results we can assume OPC can have effects in regulating wound healing process. The purpose of this study is to evaluate the effect of OPC on fibroblasts to regulate wound healing process
Materials and Methods: Human dermal fibroblast known as Hs27 cells, were treated with various concentrations of OPC (0, 2.5, 5, 10, and 20 ㎍/㎖) for 24, 48 and 72 hours. To determine cytotoxicity, cell viability was evaluated by the Cell Counting Kit assay (CCK-8
Dojindo Molecular Technologies, Rockville, MD, USA). The expression levels of intra and extra-cellular procollagen were analyzed by Western blot and real-time PCR. Gene expression of molecular chaperone and enzymes regulating collagen synthesis were determined by real-time PCR, and also intracellular localization of procollagen was determined by immunocytochemistry in OPC treated cells exposed to TGF-β1 or ascorbic acid.
Results: OPC had no proliferative effect or cytotoxicity on Hs27 cells at every concentration. The mRNA expression of procollagen, molecular chaperone and enzymes were not affected by OPC. However, OPC inhibited procollagen secretion in dose-dependent manner. The inhibitory effect also appeared under TGF-β1 induced collagen overproduction. Immunocytochemistry showed that ascorbic acid, essential cofactor in catalyzing post-translational hydroxylation of the procollagen, induced release of accumulated procollagen under OPC treatment.
Conclusion: In conclusion, OPC inhibits procollagen secretion from fibroblasts with no effects on cell proliferations. OPC has no effects on cell proliferation, and can regulate the diseases and symptoms of abnormal overabundant collagen production.