Anti-aging Effect of Brown Alga Sargassum horneri derived Chromenes in Human Dermal Fibroblasts
- Abstract
- Significant increase in the life expectancy in most countries, is closely related to the growing interest in impact of aging on the function and appearance of skin. Skin aging is influenced by several factors, especially solar UV irradiation is considered to be one of the most important causes of skin aging.
Marine algae have produced a variety of biologically active secondary metabolites. It has been reported that bioactive substances derived from marine algae are closely related to various skin symptoms. Moreover, algae and their extracts are being used as cosmetic ingredients such as antioxidants, thickening agents and moisturizing agents in a number of countries up to date. Despite these efforts, anti-photoaging effect of algal-derived compounds and their mechanism actions have not been well established. Thus, marine algae are attractive targets for the development of new cosmetic agents and skin drugs.
This research is focused on the development of natural agents for cosmeceuticals from marine brown alga, Sargassun horneri as cosmeceuticals. In the present study, we isolated three chromenes from 85% aq. MeOH fraction of Sargassum horneri, Sargachromenol, Sargachromanol E and Sargachromanol D.
Effects of three chromenes on the prevention of photoaging were investigated by measuring ROS production, cytokine release and aging related gene expression in UV-A irradiated human skin dermal fibroblasts (HDF). A number of studies demonstrated that activation of matrix metalloproteinases (MMPs) might be involved in photoaging process, but little study is known about their direct contribution to ultraviolet-mediated changes in skin dermis in vitro. The results indicated that three chromenes suppressed the collagen degrading MMPs, MMP-1, MMP-2, MMP-3 and MMP-9 expression. It was further found that these inhibitions are due to increase in expression of TIMP-1 and TIMP-2 genes. In addition, treatment with three chromenes enhanced the expression of collagen synthetic markers such as procollagen and type I collagen in UV-A irradiated dermal fibroblasts. Moreover, treatment with chromenes enhanced elastin expression by regulation of elastase activity. Since hyaluronan has been known to control cell proliferation and regeneration, effect of UV-A on the expression of hyaluronan synthases (HASs) was examined in cultured human dermal fibroblasts. HAS1, HAS2 and HAS3 mRNA were predominantly up-regulated after a single irradiation with 6 J/cm2 dose of UV-A. Increased accumulation of hyaluronan in the culture medium was proportional to the UV-A induced increase in the mRNA levels of HAS1, HAS2 and HAS3. It can be suggested that UV-A-irradiated fibroblasts may enhance hyaluronan production to maintain homeostasis through up-regulation of HAS 1, HAS2 and HAS3 genes. Furthermore, we confirmed that the UV-A induced transcriptions of AP-1 and TGF-β/Smad signaling cascade were regulated by three chromenes treatment in UV-A irradiated dermal fibroblasts.
Therefore, this study suggests that three compounds isolated from Sargassum horneri can be developed as topical application of natural anti-aging cosmeceuticals as they reduced the skin collagen decomposition by enhancement of collagen synthesis.
- Issued Date
- 2012
- Awarded Date
- 2012. 2
- Type
- Dissertation
- Publisher
- 부경대학교
- Affiliation
- 부경대학교 화학과
- Department
- 대학원 화학과
- Advisor
- 김세권
- Table Of Contents
- Table of Contents
Abstract i
Table of Contents iii
List of Figures viii
List of Tables xiii
List of Abbreviation xiv
Chapter 1. Research Background 1
Cosmeceuticals from marine resources 2
1. Algae extract 3
2. Terpenoids and Carotenoids 3
3. Polysaccharides 4
4. Polyunsatuared Fatty Acids 6
5. Proteins 7
6. Tocopherol 7
7. Phenolic Compounds 8
Chapter 2. Isolation and chracterization of chromenes from sargassum horneri 10
1. INTRODUCTIONS 11
2. MATERIALS AND METHODS 14
2.1. Instruments and Reagents 14
2.2. HPLC system 15
2.3. Extraction and isolation 15
3. RESULTS 20
3.1. Isolation of compounds from Sargassum horneri 20
3.2. Spectroscopic properties of compounds isolated from Sargassum horneri 20
3.3. Identification of compounds isolated from Sargassum horneri 23
4. SUMMARY 40
Chapter 3. Anti-photoaging effect of chromenes from sargassum horneri 41
1. INTRODUCTION 42
1.1. Morphology of UV-irradiated aged human skin 43
1.2. UV-A irradiation 44
1.3. Signaling cascade induced by UV irradiation 44
1.4. Objective and significance of the study 47
2. MATERIALS AND METHODS 49
2.1. Materials 49
2.2. Cell culture 49
2.3. UV-A irradiation 50
2.4. Cell cytotoxicity and protection of photo-damage determination using MTT assay 50
2.5. Phototoxicity determination 51
2.6. Determination of intracellular formation of reactive oxygen species (ROS) using DCFH-DA labeling 51
2.7. Membrane protein oxidation 52
2.8. Membrane lipid peroxidation 53
2.9. Measurement of Intracellular GSH 54
2.10. Measurement of cytokines production by enzyme-linked immunosorbent (ELISA) assay 54
2.11. Measurement of elastase activity 55
2.12. Collagen degrading MMPs level by enzyme-linked immunosorbent (ELISA) assay 56
2.13. Procollagen secretion determined by ELISA assay 57
2.14. Hyaluronan content determined by ELISA assay 58
2.15. Immunofluorescent labeling 59
2.16. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis 59
2.17. Western blot analysis 62
2.18. Statistical analysis 62
3. RESULTS 63
3.1. Optimization of UV-A irradiation dosage 63
3.2. Chromenes have protective effect on the viability of UV-A induced dermal fibroblasts 72
3.3. Phototoxic effect of compounds 72
3.4. Chromenes inhibited intracellular ROS generation caused by UV-A irradiation 76
3.5. Inhibition of membrane protein oxidation 80
3.6. Chromenes have protective effect on membrane lipid peroxidation caused by UV-A irradiation 82
3.7. Chromenes enhanced GSH level in UV-A irradiated dermal fibroblasts 84
3.8. Chromenes inhibit production of inflammatory cytokines by UV-A 86
3.9. Chromenes inhibited inflammatory response gene and protein 91
3.10. Chromenes inhibited UV-A induced Hyarulonan secretion 94
3.11. Chromenes inhibited mRNA expression of hyaluronan syntheases induced by UV-A irradiation 98
3.12. Chromenes inhibited elastase 100
3.13. Inhibitory effect of chromenes on UVA-induced collagen degrading MMP secretions 101
3.14. Chromenes enhanced the procollagen level 104
3.15. Chromenes suppressed UV-A-induced collagen degrading MMP expressions 106
3.16. Chromenes enhanced elastic fiber expression in UV-A irradiated fibroblasts 111
3.17. Chromenes enhanced decomposition of extracellular matrix components caused by UV-A 114
3.18. Chromenes inhibited UV-A induced AP-1 signal pathway 119
3.19. Chromenes enhanced down-regulation of TGF-β/Smad signaling by UV-A irradiation 121
3.20. Chromenes regulated AP-1 and TGF-β/Samd in UV-A dependent 123
4. DISCUSSION 126
5. SUMMARY 134
6. CONCLUSIONS 135
7. REFERENCES 137
- Degree
- Doctor
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