Protective effect of polysaccharides from Sargassum fusiforme against UVB-induced oxidative stress in HaCaT human keratinocytes
Introduction
Human skin, as the largest and visible organ of the body, which can be divided into three layers including epidermis, dermis and hypodermis (Hou et al., 2012). Skin is an effective physical and chemical barrier against the oxidative environment directly including solar ultraviolet (UV). UV can be divided into UVA (320–400 nm), UVB (290–320 nm) and UVC (100–290 nm) based on wavelength. Among these, UVC is absorbed by the ozone layer which blocked from reaching the earth’s surface (Dai et al., 2011). UV irradiation is mainly composed of UVA and UVB rays. Though UVB is a minor part of solar UV irradiation, which is 1000 times more capable of photoaging damage than UVA (Martinez et al., 2015, Matsumura and Ananthaswamy, 2004). Excessive exposure of the skin to UVB irradiation causes several pathological changes which are characterized by erythema, edema, sunburn, hyperplasia, inflammation, immune suppression, photoaging, and carcinogenesis (Martinez et al., 2015). Chronic exposure of the skin to UVB radiation could increase the generation of excessive reactive oxygen species (ROS) and decrease antioxidant levels in the skin (Fan, Zhuang, & Li, 2013). Excessive amount of ROS would break the balance of antioxidant defense systems which directly increase oxidative DNA damage and peroxidation of lipid and protein in the skin, and finally cause oxidative stress and photoaging (Lee, Ko et al., 2013, Lee, Yang et al., 2013). UVB-induced ROS mediated the epidermal growth factor and cytokines binding to the receptors which would activate the matrix metalloproteinases (MMPs) in both the epidermis and dermis (Tiraravesit et al., 2015), and then promote collagen breakdown by upregulating extracellular matrix and elastin in the dermis (Lee, Ko et al., 2013, Lee, Yang et al., 2013). Collagen, the most abundant extracellular matrix (ECM) proteins which are the main building blocks of the skin in the dermis, is derived from dermal fibroblasts and regulated by mitogen-activated protein (MAP) kinase (Halliday, 2005). MAP kinase induces activator protein-1 (AP-1; a transcription factor containing c-Fos and c-Jun) and promotes collagen breakdown by upregulating enzyme called matrix metalloproteinases (MMPs). MMPs are a family of zinc dependent enzyme related matrix-degrading which played crucial roles in skin aging and photodamage. Especially, MMP-1, known as an interstitial collagenase, is the major collagen-degrading enzyme which is related to the degradation of collagen type I, II and III in the skin (Chiang, Chan, Chu, & Wen, 2015). In addition, MMP-9 also plays vital roles in photoaging by degrading ECM in the dermis (Chiang et al., 2015). More and more scholars pay attention to the skin care for maintenance of healthy skin in recent years. Various botanical antioxidants supplement indicated protective effects against UVB-induced skin injury, such as chlorogenic acid (Cha et al., 2014), phenolic (Kim, Park, Lee, Lim, & Nho, 2015), flavonoids (Chiang et al., 2015) and collagen peptide (Fujii et al., 2013). However, little work has been done on the skin protective effect of polysaccharides from Sargassum fusiforme. Sargassum fusiforme, belonging to the family of Sargassaceae, is a kind of brown algae extensively distributed in the coastal zones in Asia (Cong et al., 2016). Sargassum fusiforme not only has been used as a Chinese health food but also as herbal medicine for thousands of years (Hu et al., 2016). Sargassum fusiforme contains many nutritional components, such as polysaccharides, protein, minerals, vitamins and dietary fiber (Zhou, Hu, Wu, Pan, & Sun, 2008). Modern pharmacological studies demonstrated that Sargassum fusiforme polysaccharides possessed multiple functions such as antitumor (Chen et al., 2012), antioxidant (Wang et al., 2013), hypolipidemic, immunomodulatoty activities (Chen, Nie, Yu et al., 2012, Hu et al., 2014). Due to these multiple biological activities, structures and properties of Sargassum fusiforme polysaccharides (SFP) have been investigated widely. However, little work has been reported on the relationships between key structural characteristics of SFP and its biological activity. Therefore, in order to explore and utilize Sargassum fusiforme resource, it is necessary to investigate the structural properties of Sargassum fusiforme polysaccharides and their biological activities. In this study, Sargassum fusiforme polysaccharides were extracted by hot water and purified using a DEAE-Sepharose fast-flow column to obtain SFP-P1 fraction. The chemical composition, preliminary structural properties and protective effects against UVB-induced oxidative stress of SFP-P1 were investigated.
Section snippets
Chemicals and reagents
Sargassum fusiforme was collected from Dongtou (Zhejiang China) in May 2015. DMEM was purchased from Gibco Biotechnology Co. (Grand Island, NY, USA). The human keratinocyte cells were obtained from Jennio Biological (Guangzhou, Guangdong, China). The assay kits for superoxidase dismutase (SOD), glutathione peroxidase (GSH-Px), and reactive oxygen species (ROS) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China). MMP-1 and MMP-9 ELISA assay kits were
Chemical compositions and structural characteristics of SFP-P1 analysis
SFP-P1 was obtained after purification using a DEAE-Sepharose fast-flow column. Chemical compositions and structural characteristics of SFP-P1 polysaccharides were as follows (Table 1): As expected, the SFP-P1 contains mainly carbohydrates (87%) with less than 1% protein. Within the carbohydrate, it also contains about 10% sulfate and 18% glucuronic acids. SFP-P1 consisted of l-fucose (13.17%), galactose (4.28%), glucose (1.95%), xylose (5.50%), and mannose (75.10%) with an average molecular
Conclusions
In this study, Sargassum fusiforme polysaccharides were purified using a DEAE-Sepharose fast flow column to obtain Sargassum fusiforme polysaccharides fraction (SFP-P1). The monosaccharide profile of SFP- P1 was l-fucose, galactose, glucose, xylose, and mannose, respectively. And the glycosidic linkages of SFP-P1 polysaccharides were proven to be →3,6)-α-d-Manp(1→, →4)-α-d-GalAp(1→, →4)-β-d-Xylp(1→ and → 3,4)-α-d-GlcAp(1→. Furthermore, SFP-P1 could against UVB-induced oxidative stress in HaCaT
Acknowledgements
This work was funded by the program of Guangdong Special Support Program (2015TQ01N670), Pearl River S&T Nova Program of Guangzhou (201610010096), the Fundamental Research Funds for the Central Universities (2015ZZ110), the National Natural Science Foundation of Guangdong Province (2014A030313242) and “Leading Talent of Guangdong Province”.
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