Hyaluronidase inhibitor derived from symbiotic fermentation product and applications of hyaluronidase inhibitor
A technology of hyaluronidase and symbiotic fermentation, which is applied in skin care preparations, medical preparations containing active ingredients, anti-inflammatory agents, etc., and can solve problems such as limited inhibitory effect
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Embodiment 1
[0048] Embodiment 1: Preparation of symbiotic fermentation product
[0049] 1. Preparation of vegetable medium
[0050] First, according to Table 1, the vegetative culture media of Examples and Comparative Examples 1 to 2 were prepared.
[0051] Table 1
[0052]
[0053] Before fermentation, the vegetable culture medium listed in Table 1 can be sterilized by a known high-pressure steam sterilization method, and the sterilization process conditions can be, for example, 121° C. for 40 minutes. Then, inoculate at least 8 different fermenting strains into the vegetative medium shown in Table 1, wherein the total inoculum of the fermenting strains in the vegetative medium can be 3% (w / v) to 5% (w / v ).
[0054] In this embodiment, the aforementioned fermentation strains include Bifidobacterium adolescentis (B.adolescentis; BCRC 14606 or ATCC 15703, and / or BCRC 14658 or DSM 20087), Bifidobacterium bifidus (B.bifidum; BCRC 11845 or ATCC 29521 , and / or BCRC 14615 or ATCC 29521),...
Embodiment 2
[0056] Example 2: Evaluation of Symbiotic Fermentation Products Using High Performance Liquid Chromatography
[0057] 1. Analytical product extraction of symbiotic fermentation products
[0058] The symbiotic fermentation products obtained in the experimental example of Example 1, Comparative Example 1, and Comparative Example 2 were first extracted with 50% ethyl acetate, and then the filtrate was filtered with multiple layers (for example, 6 layers) of gauze, and dried at 45 ° C. Dry for 6 hours to obtain the extract product of the symbiotic fermentation product test sample, take 1 mg sample, and dissolve it back in 2 mL DMSO for high performance liquid chromatography (HPLC) analysis.
[0059] 2. Analytical method for the extraction product of the symbiotic fermentation product test sample
[0060] The HPLC analysis conditions are as follows, and the results are as follows: figure 1 Shown:
[0061] (i) Analysis column: Cosmosil C18;
[0062] (ii) UV-VIS detector: Hitachi...
Embodiment 3
[0069] Example 3: Assessing the hyaluronidase inhibitory activity of symbiotic fermentation products (i.e. antiallergic activity analysis)
[0070] 1. Cell Culture
[0071]First, mouse 3T3 fibroblasts (mouse 3T3 fibroblasts; BCRC 60071) were cultured in Dulbecco's Modified Eagle's Medium (DMEM ) medium and add 3.7% (w / v) sodium bicarbonate, 0.03% (w / v) glutamic acid (L-glutamine), every milliliter contains 100 units of penicillin (penicillin) and 100 mg of streptavidin Streptomycin and other antibiotics, at 37°C, 10% CO 2 Incubator culture, generally 1 × 10 6 The cell density of each cell was seeded at 75cm 2 Petri dishes were subcultured every three to four days and replaced with fresh medium. Afterwards, about the tenth generation of 3T3 cells was taken to conduct cell experiments with different concentrations of symbiotic fermentation products.
[0072] 2. Hyaluronidase activity inhibition analysis method (hyaluronan-basedzymography analysis; hyaluronan-basedzymography...
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