Effect of Bacillus subtilis natto- fermented Radix astragali on collagen and hyaluronic acid biosynthesis in human skin cells
Date Issued
2008
Date
2008
Author(s)
Hsu, Mei-Fang
Abstract
The biochemical changes associated with skin aging include reduced expression of type I collagen messenger ribonucleic acid (mRNA), overexpression of matrix metalloproteinase (MMPs), and decreased synthesis of extracellular matrix (ECM). These changes result in increased collagen degradation and leads to skin laxity, atrophy, wrinkles, dryness, and other clinical manifestations of skin. Chinese herbs are of particular interests because they have been applied in skincare products for more than 5000 years. Radix astragali has traditionally been used for skin health, and microbial fermentation is one of the traditional methods for processing Chinese herbs. The first part of this study was to perform a screening test on the evaluation of free radical scavenging activities, collagenase and elastase inhibitory effects, and the stimulating effect on collagen production in human fibroblasts with fermented Chinese herbal preparations for potential cosmetic applications. The results revealed that the DPPH free radical scavenging effect of SAL (60.3%), at a concentration of 1250 μg/mL, was better than that of other preparations. One mg/ml of SAG and PDG were equivalent to 1.09 and 0.90 mM of Trolox in TEAC scavenging assay, respectively. In addition, 170.85 µg/mL and 369.59 µg/mL of PDG were needed for 50% collagenase and elastase inhibitory effect, respectively. The HQB treatment (500 μg/mL) increased type I procollagen production by 33% in fibroblasts (CCD-966SK) as compared with the untreated group. With relatively low toxicity and the stimulating effects on collagen production of HQB, the effect of B. subtilis natto– fermented R. astragali (HQB) on collagen and hyaluronic acid production in normal primary human skin cells were investigated in the last two parts of this research.t was found that HQNB significantly improved cell growth and proliferation of human dermal fibroblasts (HDF). Conversely, the enzyme-linked immunosorbent assay (ELISA) and Western blot analysis demonstrated that HQB, but not HQNB, significantly stimulated the biosynthesis of type I procollagen in both aged (81y) and young (22y) HDF cells in dose-dependently manners. Real-time reverse transcription-polymerase chain reaction revealed that expression of type I, type III procollagen and transforming growth factor β1 (TGF-β1) mRNA was significantly induced in HQB-treated HDF cells than that of HQNB-treated and un-treated HDF cells, suggesting that there was a close correlation between the mRNA expression and protein production in the fibroblasts response to HQB stimulation. The results also suggested that HQB could stimulate the collagen biosynthesis in human dermal fibroblasts, which is, at least in part, associated with the regulation of procollagen biosynthesis resulting from HQB-induced TGF-β1 expression and the mitogenic activity in HDF cells. On the other hand, it was found that human epidermal keratinocytes (HEK) reacted to HQNB much more sensitive than to HQB on growth-stimulating effect. In contrast, HQB significantly and dose-dependently stimulated hyaluronic acid (HA) production in cultured primary human epidermal keratinocytes and human dermal fibroblasts. In addition, HQB markedly and dose-dependently increased the expression of hyaluronan synthase 3 (HAS3) and hyaluronan synthase 2 (HAS2) mRNA in HaCaT cells and human fibroblasts, respectively. These findings suggest that the enhancement of HA synthesis in HQB-treated skin cells does not mainly depend on the basis of proliferation effect, and correlates well with the higher expression of hyaluronan synthase genes. The isoflavonoids of HQB contained lower levels of calycosin, formononetin and ononin than that of HQNB, but had significantly higher enhancement effect on the production of HA in the cultures of human epidermal keratinocytes. It is suggested that isoflavonoid aglycones or other metabolites (e.g. daidzein, equol), converted from their primary isoflavones during the fermentation, might be responsible for the skincare functions found in this study. Taken together, HQB might act to reduce the age-dependent loss of extracellular matrix components (e.g. collagen and hyaluronan), and might play a promising role in antiaging cosmetic applications.
Subjects
Radix astragali
Bacillus subtilis natto
Fermentation
Collagen
Hyaluronic acid
Human epidermal keratinocytes
Human dermal fibroblasts
SDGs
Type
thesis
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