Antioxidant and anti-aging effects of Porphyra-334 in human skin primary cells and human artificial skin tissue models = 인간 피부 세포와 인공 피부 모델에서 Porphyra-334의 항산화 및 항노화 효과

다국어 초록 (Multilingual Abstract)

Antioxidant and anti-aging effects of Porphyra-334 in human skin primary cells and human artificial skin tissue models Ultraviolet radiation (UV) causes damage on skin barrier and induces skin aging. Porphyra-334 (PPR-334) is one of the spices of mycosorprine-like amino acids (MAAs). MAAs containing PPR-334 are known as biological UV protection ingredients. This study developed a large-scale purification process to extract PPR-334 from Saccharomyces cerevisiae, confirmed the previously identified efficacy of PPR-334 in primary cells and human artificial skin tissue models on UV irradiation, and recognized its independent efficacy. We observed antioxidant effect of PPR-334 using 2,2- diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2′-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid, ABTS). PPR-334 scavenged reactivity oxygen species (ROS) and increased catalase (CAT) gene expression in human epidermal keratinocyte cells (HEKa). In both HEKa cells exposed by UVA/UVB and normal human dermal fibroblast cells (NHDF), PPR-334 suppressed gene expression of matrix metalloproteinase-1 (MMP-1). NHDF cells treated with PPR-334 showed collagen expression increased and proliferation, while advanced glycation end-products (AGEs) production was decreased. It was confirmed that the efficacy in vitro was also reproduced on human artificial skin tissue models. Above all, the mechanism of antioxidant efficacy of PPR-334 in HEKa cells through nuclear factor erythroid 2–related factor 2 (NRF2) and Caspase-9 signals was identified. It was determined that the proliferation efficacy of PPR-334 was due to factors related to the cell cycle. These results demonstrate the anti-aging efficacy of PPR-334 independent of UV irradiation, while enhancing the UV-blocking and antioxidant effects. Thus, we suggest the potential of PPR-334 as a sunscreen agent as well as dual- or multi- functional material.

목차 (Table of Contents)

• ABSTRACT = ⅰ
• TABLE OF CONTENTS = ⅲ
• LIST OF TABLES = ⅵ
• LIST OF FIGURES = ⅶ
• LIST OF ABBREVIATIONS = ⅷ
• Ⅰ. INTRODUCTION = 1
• Ⅱ. MATERIALS AND METHODS = 4
• 2.1 Purification of PPR-334 = 4
• 2.2 2,2-diphenyl-1-pocrylhydrazyl (DPPH) radical scavenging assay = 4
• 2.3 2,2’-azino-bis(3-ehthylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+∙) scavenging assay = 5
• 2.4 Cell culture = 5
• 2.5 Cell cytotoxicity assay = 6
• 2.6 H2O2-induced cell death inhibition assay = 6
• 2.7 2’7’-dichloroflurosecin diacetate (DCF-DA) assay = 7
• 2.8 UVA/B irradiation = 7
• 2.9 Quantitative reverse transcription PCR (qRT- PCR) assay = 8
• 2.10 Cell proliferation assay = 8
• 2.11 Immunocytochemistry (ICC) assay = 9
• 2.12 Anti-glycation assay = 9
• 2.13 Label-free collagen imaging = 10
• 2.14 Western blot analysis = 11
• 2.15 Statistical analysis = 12
• Ⅲ. RESULTS = 13
• 3.1 PPR-334 exhibits potent radical scavenging activity and enhance catalase (CAT) expression = 13
• 3.2 Anti-aging effect of PPR-334 via MMP-1 suppression and collagen upregulation = 20
• 3.3 PPR-334 promotes proliferation and increase growth factor expression in NHDF = 26
• 3.4 Comprehensive evaluation of PPR-334 efficacy on human artificial skin tissue models = 29
• 3.5 Mechanism of antioxidant and proliferation effect of PPR-334 in human skin cells = 33
• Ⅳ. DISCUSSION = 37
• Ⅴ. CONCLUSION = 42
• REFERENCES = 43
• APPENDIX = 52
• 국문요지 = 53