Use of caftaric acid and lactic bacterium in food supplement for regulating skin pigmentation
一种咖啡酰酒石酸、食品补充剂的技术，应用在食品制备中使用的细菌、护理皮肤的制剂、食品成分的功能等方向，能够解决局部活性剂刺激性副作用、不能完全令人满意等问题
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Embodiment 1
[0095] Embodiment 1: the hydrolysis of caffeoyl tartaric acid
[0096] Hydrolysis of caffeoyltartaric acid by 1-chlorogenic esterase
[0097] A solution of chlorogenic esterase (0.8 mg, 24 U / g, from Kikkoman, Japan) in 100 μl of phosphate buffer (50 mM, pH 7.0) was added to caffeoyl tartaric acid (0.57 mg) in 100 μl of phosphate buffer (50mM, pH 7.0) in solution. The mixture was then incubated at 37°C for 4 hours. After the reaction, the enzyme activity was terminated by heat treatment (5min, 90°C) and the mixture was centrifuged (microcon YM10, 30min, 14000g). The supernatant was analyzed by HPLC. Control reactions were run in parallel under the same reaction conditions (but without enzyme).
[0098] Hydrolysis of caffeoyltartaric acid by Lactobacillus johnsonii (La1)
[0099] Cells of Lactobacillus johnsonii (CNCM I-1225) were grown (7.0 E08 cfu / ml) and centrifuged (5000g, 10min), and the cell pellet was resuspended in phosphate buffer (50mM , pH 7.0). To 100 µl of ...
Embodiment 2
[0126] Example 2: Effect on Skin Pigmentation
[0127] To assess the potential beneficial effects of ingredients on skin depigmentation or promoting hyperpigmentation, we performed 2 experiments with 2D cultures of murine melanocytes (B16): 1 - assessing melanogenesis and 2 - assessing tyrosinase production.
[0128] 1. cell culture conditions
[0129] Incubate B16 cells in DMEM containing 1 g / L glucose without phenol red supplemented with 10% fetal bovine serum in a humidified culture room at 37 °C with 5% CO 2 cultivated under conditions.
[0130] 2. Melanin produced by the B16 murine melanocyte line
[0131] Cells were incubated with selected components or assay reference (kojic acid, 400 μg / mL) in the presence or absence of NDP-MSH, an analogue of MSH, for 72 hours. The total amount of melanin (extracellular and intracellular) was assessed by measuring the optical density at 405 nm of each sample compared to a melanin standard in the presence or absence of NDP-MSH...
Embodiment 3
[0141] Example 3: Effects on Skin Barrier Function and Skin Moisture Content
[0142] The potential beneficial effects of the extract of Example 2 on skin barrier function and skin moisture content were assessed using 2D cultures of human prodermal keratinocytes by assessing their filaggrin synthesis after treatment with selected components.
[0143] cell culture conditions
[0144] Culture human epidermal keratinocytes in restricted keratinocyte serum-free medium in a humidified culture chamber at 37 °C with 5% CO 2 cultivated under conditions.
[0145] Synthesis of filaggrin by human epidermal keratinocytes
[0146] Cells were mixed with selected components or assay references (CaCl 2 , 1.5mM) together for 144 hours. Filaggrin production was assessed by immunolabeling.
[0147] result
[0148] The result is as image 3 shown. Pretreatment of cells with caffeoyl tartaric acid + La1 showed a significant increase in filaggrin (280% of control, image 3 ), sugge...
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