Effects of Spirulina platensis polysaccharides on promotion of extracellular matrix production and inhibition of hydrogen peroxide induced fibroblasts injury
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Presented by: Dengliang Yang
The application of algae-derived ingredients in cosmetics has received more and more attention in the treatment of skin problems, such as skin dryness, skin aging and pigment disorders. Spirulina platensis is known for its higher content of protein. However, little data has been reported about the exploitation of its polysaccharides in cosmetics. In previous studies, we reported a new strain of Spirulina platensis mutant (SPm) with higher content of polysaccharides (19.60% vs 4.72%) after space breeding. In this study, we further explored the mechanism of polysaccharides from SPm in skin care.
Molecular weights of SPm polysaccharides were determined by gel-permeation chromatography (GPC). The tetrazolium dye MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell toxicity of polysaccharides to human skin fibroblasts. Enzyme-linked immunosorbent assay was used to measure the hyaluronic acid (HA) and type Ⅰ collagen (ColI) levels in the supernatants of cultured fibroblasts after the treatment with different doses of polysaccharides. Hydrogen peroxide (H2O2)-induced oxidative stress on fibroblasts was established to investigate the protective effects of polysaccharides. Biochemical methods were used to measure the levels of malondialdehyde (MDA), glutathione (GSH) and the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in H2O2-treated fibroblasts with or without the treatment of polysaccharides. Dichlorodihydrofluorescein (DCF) was used to label reactive oxygen species (ROS) and monitor its changes in fibroblasts. Apoptotic rate of H2O2-treated fibroblasts was observed using Hoechst, 33342 dye under fluorescent microscope.
The low molecular weight fractions (< 800 Da) comprised more than 20% of the SPm polysaccharides. MTT assay demonstrated the polysaccharides had no significant toxicity or proliferation promoting effects on fibroblasts when less than 160 mg/L. SPm polysaccharides could increase the levels of type I collagen and hyaluronic acid in the supernatants of cultured fibroblasts dose-dependently. Thus, those results demonstrated that SPm polysaccharides could promote the production of extracellular matrix (ECM) in fibroblasts.
H2O2 induced apoptosis and oxidative injury in fibroblasts. Pre-treatment of fibroblasts with SPm polysaccharides significantly reversed the H2O2-induced decrease of SOD and GSH-Px activity. Meanwhile, SPm polysaccharides could increase the concentration of GSH and decrease the levels of MDA and ROS in fibroblasts after H2O2 treatment. This fact demonstrated that SPm polysaccharides were effective against H2O2-induced fibroblasts injury by enhancing the activity of SOD and GSH-Px.
In summary, SPm polysaccharides could increase the secretion of ECM and protect fibroblasts from oxidative injury caused by H2O2. Our results suggest that SPm polysaccharides have the potential to be used further in cosmetics and in the prevention of skin aging and oxidation-related skin problems.
Molecular weights of SPm polysaccharides were determined by gel-permeation chromatography (GPC). The tetrazolium dye MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell toxicity of polysaccharides to human skin fibroblasts. Enzyme-linked immunosorbent assay was used to measure the hyaluronic acid (HA) and type Ⅰ collagen (ColI) levels in the supernatants of cultured fibroblasts after the treatment with different doses of polysaccharides. Hydrogen peroxide (H2O2)-induced oxidative stress on fibroblasts was established to investigate the protective effects of polysaccharides. Biochemical methods were used to measure the levels of malondialdehyde (MDA), glutathione (GSH) and the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in H2O2-treated fibroblasts with or without the treatment of polysaccharides. Dichlorodihydrofluorescein (DCF) was used to label reactive oxygen species (ROS) and monitor its changes in fibroblasts. Apoptotic rate of H2O2-treated fibroblasts was observed using Hoechst, 33342 dye under fluorescent microscope.
The low molecular weight fractions (< 800 Da) comprised more than 20% of the SPm polysaccharides. MTT assay demonstrated the polysaccharides had no significant toxicity or proliferation promoting effects on fibroblasts when less than 160 mg/L. SPm polysaccharides could increase the levels of type I collagen and hyaluronic acid in the supernatants of cultured fibroblasts dose-dependently. Thus, those results demonstrated that SPm polysaccharides could promote the production of extracellular matrix (ECM) in fibroblasts.
H2O2 induced apoptosis and oxidative injury in fibroblasts. Pre-treatment of fibroblasts with SPm polysaccharides significantly reversed the H2O2-induced decrease of SOD and GSH-Px activity. Meanwhile, SPm polysaccharides could increase the concentration of GSH and decrease the levels of MDA and ROS in fibroblasts after H2O2 treatment. This fact demonstrated that SPm polysaccharides were effective against H2O2-induced fibroblasts injury by enhancing the activity of SOD and GSH-Px.
In summary, SPm polysaccharides could increase the secretion of ECM and protect fibroblasts from oxidative injury caused by H2O2. Our results suggest that SPm polysaccharides have the potential to be used further in cosmetics and in the prevention of skin aging and oxidation-related skin problems.