A composition of anti-aging fermentation product and polypeptide and its application in preparing skin care products

Abstract

The present invention relates to the technical field of skin care products, and in particular to a composition of an anti-aging fermentation product and a polypeptide and its application in the preparation of skin care products. The invention provides Bifidobacterium longum subsp. infantis ( Bifidobacterium longum The invention discloses a composition of a fermentation filtrate of Bifidobacterium longum infantis subsp. Infantis and a microcurrent tetrapeptide. Experiments have shown that the composition prepared by the two can achieve anti-aging effects by anti-oxidation, anti-photodamage and / or inhibiting harmful bacteria. In addition, the fermentation filtrate of Bifidobacterium longum infantis subsp. Infantis and the microcurrent tetrapeptide can produce a synergistic effect. The composition can be applied to the field of cosmetics, with a wider range of efficacy, especially in the field of beauty and skin care.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and in particular to a composition of an anti-aging fermentation product and a polypeptide and application thereof in preparing skin care products. Background Art

[0002] Skin is the largest organ in the human body. Its aging is a natural phenomenon in the life process. There are many factors that lead to skin aging:

[0003] First of all, the natural aging process is inevitable. As we age, the regenerative ability of skin cells gradually weakens, leading to changes in skin quality and decreased elasticity.

[0004] Secondly, environmental factors also play an important role. External stimuli such as ultraviolet rays, pollution, and extreme temperatures can accelerate skin aging. For example, ultraviolet rays can penetrate the surface of the skin and damage the collagen and elastic fibers in the skin, causing their quantity and quality to gradually decrease, causing the skin to lose support, form wrinkles and sag. At the same time, ultraviolet rays also accelerate the loss of moisture from the skin, making the skin dry and rough. These factors work together to make the skin show obvious signs of aging, such as spots, dullness, and fine lines. In addition, ultraviolet rays also promote the generation of free radicals, which damage collagen and elastic fibers, exacerbating skin sagging and wrinkle formation.

[0005] In addition, Moraxella osloensis is involved in regulating collagen metabolism, extracellular matrix assembly and promoting cell senescence in human keratinocytes and fibroblasts, and can be used as an aging indicator bacterium.

[0006] At present, the market demand for anti-aging cosmetics is strong, which has promoted the development of product diversification. Technological innovation brings new formulas and ingredients, improving product competitiveness. With the increasing variety of anti-aging cosmetics, the market is also facing more challenges. For example, the products on the market are of varying quality, and improving quality has become a current research hotspot. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a composition of an anti-aging fermentation product and a polypeptide and its application in the preparation of skin care products.

[0008] The composition provided by the invention comprises fermentation filtrate of Bifidobacterium longum subspecies infantis and microcurrent tetrapeptide.

[0009] In the composition of the present invention, the mass ratio of the fermentation filtrate of Bifidobacterium longum subspecies infantis to the microcurrent tetrapeptide is (0.08-20): (0.08-20).

[0010] The present invention compounds the fermentation filtrate of Bifidobacterium longum infantis subspecies and microcurrent tetrapeptide, and the anti-aging effect of the obtained composition is very significant. And it has been verified that there is a synergistic effect between the two components. Especially at the mass ratio as described above, the synergistic effect is more significant. In addition, other strains were tried to be compounded with the microcurrent tetrapeptide in preliminary experiments. For example, the compounding effect of bifid yeast and microcurrent tetrapeptide is not as good as the composition as described above.

[0011] In the composition of the present invention, the preparation method of the fermentation filtrate of Bifidobacterium longum subsp. infantis comprises: inoculating and culturing Bifidobacterium longum subsp. infantis to obtain seed liquid; fermenting the seed liquid, centrifuging it, taking the supernatant, and obtaining the fermentation filtrate of Bifidobacterium longum subsp. infantis.

[0012] Preferably, in the preparation of the fermentation filtrate of Bifidobacterium longum subsp. infantis, the inoculation amount is 5%, the culture medium is MRS culture medium, and the culture conditions include anaerobic culture. The centrifugation speed is 3000 rpm and the time is 30 min. After taking the supernatant, a concentration step is also included, and the concentration multiple is 5 times.

[0013] Preferably, the Bifidobacterium longum subspecies infantis in the present invention is a strain with a deposit number of CGMCC NO.27851.

[0014] The peptide sequence of the microcurrent tetrapeptide in the composition of the present invention is LEAP.

[0015] The composition provided by the invention optimizes the mass ratio of the fermentation filtrate of Bifidobacterium longum subspecies infantis and the microcurrent tetrapeptide.

[0016] As a feasibility case, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and microcurrent tetrapeptide is (0.08~0.4):(0.08~0.15), (0.08~0.4):(0.08~0.4), (0.08~0.4):(0.08~0.8), (0.08~0.4):(0.08~2), (0.08~0.4):(0.08~10), (0.08~0.4):(0.08~20), (0.08~0.4):(0.15~0.4), (0.08~0.4):(0.15~0.8), (0.08~0.4):(0.15~2), (0.08~ 0.4): (0.15~10), (0.08~0.4): (0.15~20), (0.08~0.4): (0.4~0.8), (0.08~0.4): (0.4~2), (0.08~0.4): (0.4~10), (0.08~0.4): (0.4~20), (0.08~0.4): (0.8~2), (0.08~0.4): (0.8~10), (0.08~0.4): (0.8~20), (0.08~0.4): (2~10), (0.08~0.4): (2~20), or (0.08~0.4): (10~20).

[0017] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0018] (0.08~0.8): (0.08~0.15), (0.08~0.8): (0.08~0.4), (0.08~0.8): (0.08~0.8), (0.08~0.8): (0.08~2), (0.08~0.8): (0.08~10), (0.08~0.8): (0.08~20), (0.08~0.8): (0.15~0.4), (0.08~0.8): (0.15~0.8), (0.08~0.8): (0.15~2), (0.08~0.8): (0.15~10), ( 0.08~0.8): (0.15~20), (0.08~0.8): (0.4~0.8), (0.08~0.8): (0.4~2), (0.08~0.8): (0.4~10), (0.08~0.8): (0.4~20), (0.08~0.8): (0.8~2), (0.08~0.8): (0.8~10), (0.08~0.8): (0.8~20), (0.08~0.8): (2~10), (0.08~0.8): (2~20), or (0.08~0.8): (10~20).

[0019] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0020] (0.08~1.5): (0.08~0.15), (0.08~1.5): (0.08~0.4), (0.08~1.5): (0.08~0.8), (0.08~1.5): (0.08~2), (0.08~1.5): (0.08~10), (0.08~1.5): (0.08~20), (0.08~1.5): (0.15~0.4), (0.08~1.5): (0.15~0.8), (0.08~1.5): (0.15~2), (0.08~1.5): (0.15~10), ( 0.08~1.5): (0.15~20), (0.08~1.5): (0.4~0.8), (0.08~1.5): (0.4~2), (0.08~1.5): (0.4~10), (0.08~1.5): (0.4~20), (0.08~1.5): (0.8~2), (0.08~1.5): (0.8~10), (0.08~1.5): (0.8~20), (0.08~1.5): (2~10), (0.08~1.5): (2~20), or (0.08~1.5): (10~20).

[0021] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis to the microcurrent tetrapeptide was

[0022] (0.08~2): (0.08~0.15), (0.08~2): (0.08~0.4), (0.08~2): (0.08~0.8), (0.08~2): (0.08~2), (0.08~2): (0.08~10), (0.08~2): (0.08~20), (0.08~2): (0.15~0.4), (0.08~2): (0.15~0.8), (0.08~2): (0.15~2), (0.08~2): (0.15~10), (0.08~2): (0.15~20), (0.08~2): (0.4~0.8), (0.08~2): (0.4~2), (0.08~2): (0.4~10), (0.08~2): (0.4~20), (0.08~2): (0.8~2), (0.08~2): (0.8~10), (0.08~2): (0.8~20), (0.08~2): (2~10), (0.08~2): (2~20), or (0.08~2): (10~20).

[0023] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0024] (0.08~10): (0.08~0.15), (0.08~10): (0.08~0.4), (0.08~10): (0.08~0.8), (0.08~10): (0.08~2), (0.08~10): (0.08~10), (0.08~10): (0.08~20), (0.08~10): (0.15~0.4), (0.08~10): (0.15~0.8), (0.08~10): (0.15~2), (0.08~10): (0.15~10), ( 0.08~10): (0.15~20), (0.08~10): (0.4~0.8), (0.08~10): (0.4~2), (0.08~10): (0.4~10), (0.08~10): (0.4~20), (0.08~10): (0.8~2), (0.08~10): (0.8~10), (0.08~10): (0.8~20), (0.08~10): (2~10), (0.08~10): (2~20), or (0.08~10): (10~20).

[0025] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0026] (0.08~20): (0.08~0.15), (0.08~20): (0.08~0.4), (0.08~20): (0.08~0.8), (0.08~20): (0.08~2), (0.08~20): (0.08~10), (0.08~20): (0.08~20), (0.08~20): (0.15~0.4), (0.08~20): (0.15~0.8), (0.08~20): (0.15~2), (0.08~20): (0.15~10), ( 0.08~20): (0.15~20), (0.08~20): (0.4~0.8), (0.08~20): (0.4~2), (0.08~20): (0.4~10), (0.08~20): (0.4~20), (0.08~20): (0.8~2), (0.08~20): (0.8~10), (0.08~20): (0.8~20), (0.08~20): (2~10), (0.08~20): (2~20), or (0.08~20: (10~20).

[0027] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0028] (0.4~0.8): (0.08~0.15), (0.4~0.8): (0.08~0.4), (0.4~0.8): (0.08~0.8), (0.4~0.8): (0.08~2), (0.4~0.8): (0.08~10), (0.4~0.8): (0.08~20), (0.4~0.8): (0.15~0.4), (0.4~0.8): (0.15~0.8), (0.4~0.8): (0.15~2), (0.4~0.8): (0.15~10), ( 0.4~0.8): (0.15~20), (0.4~0.8): (0.4~0.8), (0.4~0.8): (0.4~2), (0.4~0.8): (0.4~10), (0.4~0.8): (0.4~20), (0.4~0.8): (0.8~2), (0.4~0.8): (0.8~10), (0.4~0.8): (0.8~20), (0.4~0.8): (2~10), (0.4~0.8): (2~20), or (0.4~0.8): (10~20).

[0029] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0030] (0.4~1.5): (0.08~0.15), (0.4~1.5): (0.08~0.4), (0.4~1.5): (0.08~0.8), (0.4~1.5): (0.08~2), (0.4~1.5): (0.08~10), (0.4~1.5): (0.08~20), (0.4~1.5): (0.15~0.4), (0.4~1.5): (0.15~0.8), (0.4~1.5): (0.15~2), (0.4~1.5): (0.15~10), ( 0.4~1.5): (0.15~20), (0.4~1.5): (0.4~0.8), (0.4~1.5): (0.4~2), (0.4~1.5): (0.4~10), (0.4~1.5): (0.4~20), (0.4~1.5): (0.8~2), (0.4~1.5): (0.8~10), (0.4~1.5): (0.8~20), (0.4~1.5): (2~10), (0.4~1.5): (2~20), or (0.4~1.5): (10~20).

[0031] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0032] (0.4~2): (0.08~0.15), (0.4~2): (0.08~0.4), (0.4~2): (0.08~0.8), (0.4~2): (0.08~2), (0.4~2): (0.08~10), (0.4~2): (0.08~20), (0.4~2): (0.15~0.4), (0.4~2): (0.15~0.8), (0.4~2): (0.15~2), (0.4~2): (0.15~10), (0.4~2): (0.15~20), (0.4~2): (0.4~0.8), (0.4~2): (0.4~2), (0.4~2): (0.4~10), (0.4~2): (0.4~20), (0.4~2): (0.8~2), (0.4~2): (0.8~10), (0.4~2): (0.8~20), (0.4~2): (2~10), (0.4~2): (2~20), or (0.4~2): (10~20).

[0033] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0034] (0.4~10): (0.08~0.15), (0.4~10): (0.08~0.4), (0.4~10): (0.08~0.8), (0.4~10): (0.08~2), (0.4~10): (0.08~10), (0.4~10): (0.08~20), (0.4~10): (0.15~0.4), (0.4~10): (0.15~0.8), (0.4~10): (0.15~2), (0.4~10): (0.15~10), (0.4~10): (0.15~20), (0.4~10): (0.4~0.8), (0.4~10): (0.4~2), (0.4~10): (0.4~10), (0.4~10): (0.4~20), (0.4~10): (0.8~2), (0.4~10): (0.8~10), (0.4~10): (0.8~20), (0.4~10): (2~10), (0.4~10): (2~20), or (0.4~10: (10~20).

[0035] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0036] (0.4~20): (0.08~0.15), (0.4~20): (0.08~0.4), (0.4~20): (0.08~0.8), (0.4~20): (0.08~2), (0.4~20): (0.08~10), (0.4~20): (0.08~20), (0.4~20): (0.15~0.4), (0.4~20): (0.15~0.8), (0.4~20): (0.15~2), (0.4~20): (0.15~10), (0.4~20): (0.15~20), (0.4~20): (0.4~0.8), (0.4~20): (0.4~2), (0.4~20): (0.4~10), (0.4~20): (0.4~20), (0.4~20): (0.8~2), (0.4~20): (0.8~10), (0.4~20): (0.8~20), (0.4~20): (2~10), (0.4~20): (2~20), or (0.4~20: (10~20).

[0037] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0038] (0.8~1.5): (0.08~0.15), (0.8~1.5): (0.08~0.4), (0.8~1.5): (0.08~0.8), (0.8~1.5): (0.08~2), (0.8~1.5): (0.08~10), (0.8~1.5): (0.08~20), (0.8~1.5): (0.15~0.4), (0.8~1.5): (0.15~0.8), (0.8~1.5): (0.15~2), (0.8~1.5): (0.15~10), ( 0.8~1.5): (0.15~20), (0.8~1.5): (0.4~0.8), (0.8~1.5): (0.4~2), (0.8~1.5): (0.4~10), (0.8~1.5): (0.4~20), (0.8~1.5): (0.8~2), (0.8~1.5): (0.8~10), (0.8~1.5): (0.8~20), (0.8~1.5): (2~10), (0.8~1.5): (2~20), or (0.8~1.5): (10~20).

[0039] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0040] (0.8~2): (0.08~0.15), (0.8~2): (0.08~0.4), (0.8~2): (0.08~0.8), (0.8~2): (0.08~2), (0.8~2): (0.08~10), (0.8~2): (0.08~20), (0.8~2): (0.15~0.4), (0.8~2): (0.15~0.8), (0.8~2): (0.15~2), (0.8~2): (0.15~10), (0.8~2): (0.15~20), (0.8~2): (0.4~0.8), (0.8~2): (0.4~2), (0.8~2): (0.4~10), (0.8~2): (0.4~20), (0.8~2): (0.8~2), (0.8~2): (0.8~10), (0.8~2): (0.8~20), (0.8~2): (2~10), (0.8~2): (2~20), or (0.8~2): (10~20).

[0041] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0042] (0.8~10): (0.08~0.15), (0.8~10): (0.08~0.4), (0.8~10): (0.08~0.8), (0.8~10): (0.08~2), (0.8~10): (0.08~10), (0.8~10): (0.08~20), (0.8~10): (0.15~0.4), (0.8~10): (0.15~0.8), (0.8~10): (0.15~2), (0.8~10): (0.15~10), (0.8~10): (0.15~20), (0.8~10): (0.4~0.8), (0.8~10): (0.4~2), (0.8~10): (0.4~10), (0.8~10): (0.4~20), (0.8~10): (0.8~2), (0.8~10): (0.8~10), (0.8~10): (0.8~20), (0.8~10): (2~10), (0.8~10): (2~20), or (0.8~10: (10~20).

[0043] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0044] (0.8~20): (0.08~0.15), (0.8~20): (0.08~0.4), (0.8~20): (0.08~0.8), (0.8~20): (0.08~2), (0.8~20): (0.08~10), (0.8~20): (0.08~20), (0.8~20): (0.15~0.4), (0.8~20): (0.15~0.8), (0.8~20): (0.15~2), (0.8~20): (0.15~10), (0.8~20): (0.15~20), (0.8~20): (0.4~0.8), (0.8~20): (0.4~2), (0.8~20): (0.4~10), (0.8~20): (0.4~20), (0.8~20): (0.8~2), (0.8~20): (0.8~10), (0.8~20): (0.8~20), (0.8~20): (2~10), (0.8~20): (2~20), or (0.8~20: (10~20).

[0045] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0046] (1.5~2): (0.08~0.15), (1.5~2): (0.08~0.4), (1.5~2): (0.08~0.8), (1.5~2): (0.08~2), (1.5~2): (0.08~10), (1.5~2): (0.08~20), (1.5~2): (0.15~0.4), (1.5~2): (0.15~0.8), (1.5~2): (0.15~2), (1.5~2): (0.15~10), (1.5~2): (0.15~20), (1.5~2): (0.4~0.8), (1.5~2): (0.4~2), (1.5~2): (0.4~10), (1.5~2): (0.4~20), (1.5~2): (0.8~2), (1.5~2): (0.8~10), (1.5~2): (0.8~20), (1.5~2): (2~10), (1.5~2): (2~20), or (1.5~2): (10~20).

[0047] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0048] (1.5~10): (0.08~0.15), (1.5~10): (0.08~0.4), (1.5~10): (0.08~0.8), (1.5~10): (0.08~2), (1.5~10): (0.08~10), (1.5~10): (0.08~20), (1.5~10): (0.15~0.4), (1.5~10): (0.15~0.8), (1.5~10): (0.15~2), (1.5~10): (0.15~10), (1.5~10): (0.15~20), (1.5~10): (0.4~0.8), (1.5~10): (0.4~2), (1.5~10): (0.4~10), (1.5~10): (0.4~20), (1.5~10): (0.8~2), (1.5~10): (0.8~10), (1.5~10): (0.8~20), (1.5~10): (2~10), (1.5~10): (2~20), or (1.5~10: (10~20).

[0049] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0050] (1.5~20): (0.08~0.15), (1.5~20): (0.08~0.4), (1.5~20): (0.08~0.8), (1.5~20): (0.08~2), (1.5~20): (0.08~10), (1.5~20): (0.08~20), (1.5~20): (0.15~0.4), (1.5~20): (0.15~0.8), (1.5~20): (0.15~2), (1.5~20): (0.15~10), (1.5~20): (0.15~20), (1.5~20): (0.4~0.8), (1.5~20): (0.4~2), (1.5~20): (0.4~10), (1.5~20): (0.4~20), (1.5~20): (0.8~2), (1.5~20): (0.8~10), (1.5~20): (0.8~20), (1.5~20): (2~10), (1.5~20): (2~20), or (1.5~20: (10~20).

[0051] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0052] (2~10): (0.08~0.15), (2~10): (0.08~0.4), (2~10): (0.08~0.8), (2~10): (0.08~2), (2~10): (0.08~10), (2~10): (0.08~20), (2~10): (0.15~0.4), (2~10): (0.15~0.8), (2~10): (0.15~2), (2~10): (0.15~10) , (2~10): (0.15~20), (2~10): (0.4~0.8), (2~10): (0.4~2), (2~10): (0.4~10), (2~10): (0.4~20), (2~10): (0.8~2), (2~10): (0.8~10), (2~10): (0.8~20), (2~10): (2~10), (2~10): (2~20), or (2~10: (10~20).

[0053] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0054] (2~20): (0.08~0.15), (2~20): (0.08~0.4), (2~20): (0.08~0.8), (2~20): (0.08~2), (2~20): (0.08~10), (2~20): (0.08~20), (2~20): (0.15~0.4), (2~20): (0.15~0.8), (2~20): (0.15~2), (2~20): (0.15~10) 、(2~20):(0.15~20), (2~20):(0.4~0.8), (2~20):(0.4~2), (2~20):(0.4~10), (2~20):(0.4~20), (2~20):(0.8~2), (2~20):(0.8~10), (2~20):(0.8~20), (2~20):(2~10), (2~20):(2~20), or (2~20:(10~20);

[0055] In other cases, the mass ratio of the fermentation filtrate of Bifidobacterium infantis subsp. infantis and the microcurrent tetrapeptide is:

[0056] (10~20): (0.08~0.15), (10~20): (0.08~0.4), (10~20): (0.08~0.8), (10~20): (0.08~2), (10~20): (0.08~10), (10~20): (0.08~20), (10~20): (0.15~0.4), (10~20): (0.15~0.8), (10~20): (0.15~2), (10~20): (0.15~10), (10~20): (0.15~20), (10~20): (0.4~0.8), (10~20): (0.4~2), (10~20): (0.4~10), (10~20): (0.4~20), (10~20): (0.8~2), (10~20): (0.8~10), (10~20): (0.8~20), (10~20): (2~10), (10~20): (2~20), or (10~20: (10~20).

[0057] Preferably, the mass ratio of the fermentation filtrate of Bifidobacterium longum infantis subspecies and the microcurrent tetrapeptide is 1:10 to 10:1. For example, the mass ratio of the fermentation filtrate of Bifidobacterium longum infantis subspecies and the microcurrent tetrapeptide is 1:10, 1:1 or 10:1. In addition to the above ratios, the fermentation filtrate and the microcurrent tetrapeptide also have anti-aging effects at other mass ratios, but the effect is not as good as the aforementioned ratio.

[0058] Furthermore, the present invention also provides the use of the above-mentioned composition in the preparation of anti-aging products.

[0059] In the present invention, the anti-aging includes anti-oxidation, anti-photodamage and / or inhibition of harmful bacteria.

[0060] Oxidation is one of the main causes of aging. In some embodiments, the anti-oxidation comprises scavenging free radicals. In some specific embodiments, the free radical is a DPPH free radical.

[0061] Some harmful bacteria are also one of the causes of aging. In some embodiments, the anti-oxidation includes inhibiting harmful bacteria. In some specific embodiments, the harmful bacteria is Moraxella osloensis.

[0062] During the aging process, especially aging caused by light damage, the levels of some key genes and / or proteins also change. In the present invention, the resistance to light damage includes regulating the expression level of related proteins and regulating the transcription level of related genes.

[0063] In some embodiments, the related gene is COL3, COL 1, MMP3 and / or MMP1; in some specific embodiments, the regulation includes increasing the transcription level of COL 1 gene and / or COL3 gene, and decreasing the transcription level of MMP1 gene and / or MMP3 gene;

[0064] In some embodiments, the related protein is type I collagen and / or matrix metalloproteinases; in some specific embodiments, the regulation includes increasing the level of type I collagen and / or matrix metalloproteinases.

[0065] Aging caused by light damage often leads to cell damage. The anti-aging of the present invention, especially anti-aging caused by light damage, includes improving the ability of cells to resist ultraviolet damage; in some specific embodiments, the cells are fibroblasts.

[0066] In the embodiment of the present invention, the light damage is damage caused by ultraviolet rays. In some specific embodiments, the ultraviolet damage is damage caused by UVA.

[0067] Furthermore, the present invention also provides an anti-aging product, which comprises the composition as described above.

[0068] In the present invention, in the anti-aging product, the concentration of the fermentation filtrate of Bifidobacterium longum subsp. infantis is 0.08mg / ml~20mg / ml, or its mass fraction is 0.008%~2%. For example, the concentration of the fermentation filtrate of Bifidobacterium longum subsp. infantis is: 0.08mg / ml~0.4mg / ml, 0.08mg / ml~0.8mg / ml, 0.08mg / ml~1.5mg / ml, 0.08mg / ml~2mg / ml, 0.08mg / ml~10mg / ml, 0.08mg / ml~20mg / ml, 0.4mg / ml~0.8mg / ml, 0.4mg / ml~1.5mg / ml, 0.4mg / ml~2mg / ml, 0.4mg / 10mg / ml, 0.4mg / ml-20mg / ml, 0.8mg / ml-1.5mg / ml, 0.8mg / ml-2mg / ml, 0.8mg / ml-10mg / ml, 0.8mg / ml-20mg / ml, 1.5mg / ml-2mg / ml, 1.5mg / ml-10mg / ml, 1.5mg / ml-20mg / ml, 2mg / ml-10mg / ml, 2mg / ml-20mg / ml or 2mg / ml-20mg / ml. More specifically, the concentration of the fermentation filtrate of Bifidobacterium longum subsp. infantis is 0.08mg / ml, 0.4mg / ml, 0.8mg / ml, 1.5 mg / ml, 2mg / ml, 10mg / ml or 20mg / ml.

[0069] In the present invention, in the anti-aging product, the concentration of the microcurrent tetrapeptide is 0.08mg / ml~20mg / ml, or its mass fraction is 0.008%~2%. For example, the concentration of the microcurrent tetrapeptide is: 0.08mg / ml~0.15mg / ml, 0.08mg / ml~0.4mg / ml, 0.08mg / ml~0.8mg / ml, 0.08mg / ml~2mg / ml, 0.08mg / ml~10mg / ml, 0.08mg / ml~20mg / ml, 0.15mg / ml~0.4mg / ml, 0.15mg / ml~0.8mg / ml, 0.15mg / ml~2mg / ml, 0.15mg / ml~ 10mg / ml, 0.15mg / ml~20mg / ml, 0.4mg / ml~0.8mg / ml, 0.4mg / ml~2mg / ml, 0.4mg / ml~10mg / ml, 0.4mg / ml~20mg / ml, 0.8mg / ml~2mg / ml, 0.8mg / ml~10mg / ml, 0.8mg / ml~20mg / ml, 2mg / ml~10mg / ml, 2mg / ml~20mg / ml or 10mg / ml~20mg / ml. More specifically, the concentration of microcurrent tetrapeptide is: 0.08mg / ml, 0.15mg / ml, 0.4mg / ml, 0.8mg / ml, 2mg / ml, 10mg / ml or 20mg / ml.

[0070] In the present invention, the product is a cosmetic, which also includes cosmetically acceptable excipients.

[0071] In some embodiments, the cosmetics are cleansing cosmetics, care cosmetics or beauty / decorative cosmetics, and their dosage forms include creams, lotions, solutions, sprays, facial masks, essence water, gels or powders.

[0072] In some embodiments, the acceptable excipients in the cosmetics include, but are not limited to, purified water, oils, lipids, esters, waxes, surfactants, moisturizers, thickeners, chelating agents, pigments, fragrances, antioxidants, preservatives, pH adjusters, and the like.

[0073] The present invention also provides an anti-aging method, which comprises applying the composition and / or the product as described above on the human body surface. The method described in the present invention includes a method for therapeutic purposes and a method for non-therapeutic purposes. Preferably, it is an anti-aging method for non-therapeutic purposes. For example, a cosmetic method that does not involve the human body or produce trauma.

[0074] The invention provides a composition of a fermentation filtrate of Bifidobacterium longum subsp. Infantis and a microcurrent tetrapeptide. Experiments show that the composition prepared by the two achieves an anti-aging effect by resisting oxidation, resisting light damage and / or inhibiting harmful bacteria. In addition, the fermentation filtrate of Bifidobacterium longum subsp. Infantis and the microcurrent tetrapeptide can produce a synergistic effect. The composition can be applied to the field of cosmetics, has a wider range of efficacy, and has better performance in the field of beauty and skin care. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] Figure 1 DPPH radical scavenging rate of filtrates with different concentrations;

[0076] Figure 2 DPPH free radical scavenging rate of microcurrent tetrapeptide at different concentrations;

[0077] Figure 3 DPPH radical scavenging rate of different ratio combinations of filtrate and microcurrent tetrapeptide;

[0078] Figure 4 Cell viability of filtrates at different concentrations;

[0079] Figure 5 Cell survival rate of different concentrations of microcurrent tetrapeptide;

[0080] Figure 6 Cell viability of different ratio combinations of filtrate and microcurrent tetrapeptide;

[0081] Figure 7 Relative mRNA levels of MMP1 gene in filtrates of different concentrations;

[0082] Figure 8 Relative mRNA levels of MMP3 gene in filtrates of different concentrations;

[0083] Fig. 9 Relative mRNA levels of COL1 gene in filtrates of different concentrations;

[0084] Fig.10 Relative mRNA levels of COL3 gene in filtrates of different concentrations;

[0085] Fig.11 Relative mRNA levels of MMP1 gene at different concentrations of microcurrent tetrapeptide;

[0086] Fig.12 Relative mRNA levels of MMP3 gene at different concentrations of microcurrent tetrapeptide;

[0087] Fig.13Relative mRNA levels of COL1 gene at different concentrations of microcurrent tetrapeptide;

[0088] Fig.14 Relative mRNA levels of COL3 gene at different concentrations of microcurrent tetrapeptide;

[0089] Fig.15 Relative mRNA levels of MMP1 gene in different ratio combinations of filtrate and microcurrent tetrapeptide;

[0090] Fig.16 Relative mRNA levels of MMP3 gene in different ratio combinations of filtrate and microcurrent tetrapeptide;

[0091] Fig.17 Relative mRNA levels of COL1 gene in different ratio combinations of filtrate and microcurrent tetrapeptide;

[0092] Fig.18 Relative mRNA levels of COL3 gene in different ratio combinations of filtrate and microcurrent tetrapeptide;

[0093] Fig.19 Relative expression of MMP1 protein in the filtrate, microcurrent tetrapeptide, and the combination;

[0094] Fig. 20 Relative expression of COL1 protein in the filtrate, microcurrent tetrapeptide, and the combination;

[0095] Fig.21 Relative growth rate of bacterial flora of filtrate, microcurrent tetrapeptide and combination. DETAILED DESCRIPTION

[0096] The present invention provides a composition of an anti-aging fermentation product and a polypeptide and its application in the preparation of skin care products. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0097] Unless otherwise defined herein, scientific and technical terms related to the present invention shall have the meanings understood by those of ordinary skill in the art.

[0098] In the present invention, "include", "comprising" and "having" are used interchangeably, and are intended to indicate the inclusiveness of the scheme, meaning that the scheme may have other elements besides the listed elements. At the same time, it should be understood that the descriptions of "include", "comprising" and "having" used in this article also provide "consisting of..." schemes.

[0099] “And / or” as used herein includes the meanings of “and,” “or,” and “all or any other combination of elements linked by the term.”

[0100] In the present invention, "at least one" means one or more, "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0101] It should be understood that in various embodiments of the present invention, the size of the serial number does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0102] The numerical ranges and parameters involved in the present invention have been presented in the specific embodiments as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual testing methods. Therefore, unless otherwise explicitly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have certain reasonable deviations within a certain range, for example: within ±10%, ±5%, ±1% or ±0.5%.

[0103] Some cases are recorded in the embodiments and comparative examples of the present invention. The embodiments show some implementation methods of the present invention. However, this does not mean that the effects of the present invention can only be achieved in these cases. In fact, any concentration of each component between the two endpoint values ​​shown in the embodiments can achieve a good anti-aging effect. In the case with better effects, the mass ratio of fermentation filtrate to microcurrent tetrapeptide is 1:10, 1:1, and 10:1. The following embodiments also list some cases with poor results in the experiments, such as using different components, different ratios, and different strains. However, the effects of these attempts are not as good as the effects under the ratios described above, and will not be repeated here.

[0104] In the embodiment of the present invention, the statistical analysis method is to use GraphPad Prism 9 software for statistical analysis and drawing. If the data meet the normal distribution and variance homogeneity, the paired t test is used for the comparison before and after the experiment within the group, and the one-way analysis of variance is used for the comparison of the means between the groups. P < 0.05 indicates that the difference is statistically significant.

[0105] In the embodiments of the present invention, the synergistic analysis adopts the Bliss independence model. The Bliss independence model is used to simulate the effect of a single drug in combination, and is a common and reliable model for calculating drug synergy coefficients in the field. For related literature, see Wei Zhao et al. "A New Bliss Independence Model to Analyze Drug Combination Data" or CI BLISS "The toxicity of poisons applied jointly". The Bliss synergy score is calculated according to the Bliss independence model, and the formula is S BLISS =E A,B -(E A +E B -E A E B ), where S BLISS <0 is antagonistic, S BLISS =0 is additive effect, SBLISS >0 indicates synergistic effect.

[0106] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0107] The main component of the microcurrent tetrapeptide is Tetrapeptide-1, the peptide sequence of which is LEAP, which is purchased from Lipotec.

[0108] Bifidobacterium longum subsp. Infantis YSGBifido001 was deposited at the General Microbiology Center of China Culture Collection Administration on July 10, 2023; address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; Deposit number: CGMCCNo.27851.

[0109] The preparation of the fermentation filtrate of Bifidobacterium longum subsp. infantis includes (referring to Chinese patent application number 202410036419.6):

[0110] (1) Preparation of glycerol tube strains

[0111] 1) Activation of the original lyophilized powder: transfer the lyophilized powder to 100 mL of Huankai MRS triangular flask culture medium, and culture at 37℃ for 20-24 hours to prepare bacterial solution for use;

[0112] 2) Isolate single colonies: Streak the bacterial solution on MRS solid culture medium plates, culture at 37℃ for 48 hours, and keep them for later use after they grow up;

[0113] 3) Activation of single colony: transfer single colony to 100 mL liquid Erlenmeyer flask culture medium, and activate the bacterium-making liquid by anaerobic static culture at 37℃ for 20 hours. Check under a microscope to see if OD*10>0.260 and pH<4.50 meet the standards;

[0114] 4) Preservation of original glycerol tube strains: 5% inoculation of the above bacteria (OD*10>0.260 after 20-24 hours of culture can be considered qualified) is transferred to the culture medium in the triangular flask of formula card 1, and OD*10>0.260 and pH<4.50 are detected after 20 hours of culture. After the microscopic examination meets the standards, the strains are preserved in glycerol tubes (0.70 mL of preservative solution and 0.70 mL of bacterial solution).

[0115] 5) Establishment of seed batches: The seed batch system for production strains should be established in accordance with the relevant provisions of "Management and Quality Control of Strains for Production and Inspection of Biological Products".

[0116] (2) The strain Bifidobacterium longum subsp. infantis obtained in step (1) was inoculated into the culture medium with an inoculum amount of 5% (the culture was qualified if OD*10>0.260 was detected after 20-24 hours of culture), and cultured at 37°C for 20-24 hours. The supernatant was obtained by centrifugation and freeze-dried to obtain a concentrated fermentation liquid;

[0117] The culture medium (Formula 1) involved in the previous preparation method includes: 12 g / L lactose, 10 g / L wheat peptone, 10 g / L yeast powder, 2 g / L sodium acetate, 2 g / L dipotassium hydrogen phosphate, 2 g / L triammonium citrate, 0.58 g / L magnesium sulfate 7hydrate, 0.05 g / L manganese sulfate and 2 g / L Tween 80.

[0118] The present invention will be further described below in conjunction with embodiments:

[0119] Example

[0120] 1. DPPH free radical scavenging rate test to determine the antioxidant effect

[0121] 1. Determine the DPPH free radical scavenging rate of the sample

[0122] (1) Preparation of stock solution: 95% ethanol: Take 95 ml of ethanol in a measuring cylinder and add ultrapure water to 100 ml. 0.03% DPPH stock solution: Weigh 15 mg of DPPH powder into a 50 ml brown volumetric flask, add about 40 ml of 95% ethanol, sonicate to completely dissolve it, dilute to the mark, shake well, and use as DPPH stock solution.

[0123] (2) Preparation of 0.03‰ DPPH working solution: Based on the required amount of DPPH solution, measure 1 / 10 volume of 0.03% DPPH stock solution and dilute it to prepare 0.03‰ DPPH working solution.

[0124] (3) The positive control Trolex was diluted to 0.025 mM with pure water to verify the test system.

[0125] (4) The test substance is diluted with pure water to form samples of multiple concentrations. Referring to Table 1, set up sample tubes (T), sample background (T0), DPPH tubes (C) and solvent background (C0) in a 96-well plate. Three parallel wells should be set up for each sample well (T) of each test concentration of each sample, and three parallel wells should also be set up for the DPPH tube (C).

[0126] (5) Add 50 μl of sample solution of the same concentration and 100 μl of pure water to the sample tube (T) and sample background (T0), and add 150 μl of pure water to the DPPH tube (C) and solvent background (C0).

[0127] (6) Add 50 μl of DPPH ethanol solution to each of the sample tube (T) and DPPH tube (C). Replace the sample background (T0) and solvent background (C0) with 50 μl of 95% ethanol.

[0128] (7) Let the reaction stand at room temperature for 5 minutes and measure OD 517 .

[0129] (8) Calculate the DPPH free radical scavenging rate: scavenging rate (%) = (1- )×100%

[0130] (9) In the formula, T is the absorbance of the sample well, i.e., the absorbance of the solution after the sample reacts with DPPH; T0 is the background absorbance of the sample; C is the average of the three absorbances of the DPPH well, i.e., the absorbance of the DPPH solution when no sample is added; and C0 is the background absorbance of the solvent.

[0131] (10) Statistical analysis and graphing: GraphPad Prism 9 software was used for statistical analysis and graphing. If the data met the normal distribution and homogeneity of variance, paired t-test was used for pre- and post-experimental comparison within the group, and one-way analysis of variance was used for comparison of means between groups. P < 0.05 indicated statistically significant differences.

[0132] Table 1 DPPH free radical scavenging rate test sample solution table

[0133]

[0134] The test results are shown in Tables 2 to 4:

[0135] 1.1 Antioxidant efficacy of filtrate (DPPH free radical scavenging rate)

[0136] Table 2 DPPH free radical scavenging rate of filtrates at different concentrations

[0137]

[0138] Summary: The positive standard 0.025mM Trolex showed good DPPH free radical scavenging rate, indicating that the experimental system was successful. The filtrates of different concentrations showed excellent DPPH free radical scavenging rates, and had good concentration dependence. The DPPH free radical scavenging rate of the 10mg / ml filtrate reached 50%, indicating that the filtrate had strong antioxidant capacity and strong anti-aging effect.

[0139] 1.2 Antioxidant effect of microcurrent tetrapeptide (DPPH free radical scavenging rate)

[0140] Table 3 DPPH free radical scavenging rate of microcurrent tetrapeptide at different concentrations

[0141]

[0142] Summary: The positive standard 0.025mM Trolex showed good DPPH free radical scavenging rate, indicating that the experimental system was successful. However, different concentrations of microcurrent tetrapeptide did not show good DPPH free radical scavenging rate, indicating that microcurrent tetrapeptide has no antioxidant ability.

[0143] 1.3 Antioxidant efficacy of filtrate + microcurrent tetrapeptide in different ratios (DPPH free radical scavenging rate)

[0144] Table 4 DPPH free radical scavenging rate of different ratios of filtrate and microcurrent tetrapeptide

[0145]

[0146] Summary: When different ratios of filtrate and microcurrent tetrapeptide were combined, there was no significant difference in DPPH free radical scavenging rate compared with the filtrate alone. Only when the filtrate and microcurrent tetrapeptide were combined at a ratio of 10:1, the scavenging rate was slightly higher than that of the filtrate alone. This indicates that the filtrate and microcurrent tetrapeptide did not synergistically improve the antioxidant capacity and anti-aging effect in different combinations.

[0147] 2. MTT assay to determine cell viability

[0148] (1) HFF fibroblasts were plated into 96-well plates.

[0149] (2) On the second day, the culture medium was replaced with PBS solution and the cells were subjected to 30 J / cm 2 After UVA irradiation, fresh pre-warmed culture medium was replaced and the test samples were then added to stimulate the cells for 24 h. The final culture medium volume in each well was 200 μl.

[0150] (3) Discard the culture medium and add MTT solution (0.5 mg / mL) to each well at 37°C and incubate for 4 hours. Discard the culture medium and add 150 µl DMSO to dissolve the precipitate. Measure the absorbance at OD 490 nm using a multifunctional microplate reader and calculate the cell viability percentage using the following formula: Cell viability (%) = 100% × (OD sample - ODctl) / ODctl

[0151] 2.1 Filtrate against UVA damage (MTT)

[0152] Table 5 Numerical table of cell viability of filtrates of different concentrations

[0153]

[0154] Summary: After UVA treatment, the cell viability decreased significantly, indicating that UVA stimulation caused damage to the cell survival rate. On this basis, after treatment with different concentrations of filtrate, the detected cell viability increased significantly compared with UVA treatment alone, indicating that the filtrate has the effect of resisting cell damage caused by UVA and has anti-aging effects.

[0155] 2.2 Microcurrent Tetrapeptide Anti-UVA Damage (MTT)

[0156] Table 6 Numerical table of cell survival rates of different concentrations of microcurrent tetrapeptide

[0157]

[0158] Summary: After UVA treatment, the cell viability decreased significantly, indicating that UVA stimulation caused damage to the cell survival rate. On this basis, after treatment with different concentrations of microcurrent tetrapeptide, the detected cell viability increased to a certain extent compared with UVA treatment alone, indicating that microcurrent tetrapeptide has the effect of resisting cell damage caused by UVA and has anti-aging effects.

[0159] 2.3 Different ratios of filtrate + microcurrent tetrapeptide synergistically resist UVA damage (MTT)

[0160] Table 7 Cell survival rate values ​​of different ratios of filtrate and microcurrent tetrapeptide combination

[0161]

[0162] UVA stimulation caused damage to the survival rate of cells, and the anti-aging model was successfully established; under UVA stimulation, the filtrate, microcurrent tetrapeptide and combination were used to treat and count the cell survival rate. The results showed that when all filtrate: microcurrent tetrapeptide combinations were combined in different proportions, the cell survival rate increased significantly compared with the single filtrate and microcurrent tetrapeptide, and S BLISS>0, indicating that the combination of filtrate and microcurrent tetrapeptide can synergistically resist the damage of UVA to cell survival rate and has a synergistic anti-aging effect.

[0163] 3. Detection and analysis of anti-aging genes (qPCR)

[0164] (1) HFF fibroblasts grown in 12-well plates were stimulated with UVA to establish the model.

[0165] (2) 24 hours after modeling, cells were collected.

[0166] (3) Use an RNA total extraction kit to extract cellular RNA, and use a reverse transcription kit to reverse transcribe and synthesize cDNA.

[0167] (4) Amplification was performed using SYBR Green qPCR Master Mix fluorescent quantitative PCR instrument, and the specificity of the primers was checked by melting curve.

[0168] (5) The relative expression levels of the target genes (type I matrix metalloproteinase MMP1, type III matrix metalloproteinase MMP3, type I collagen COL1, and type III collagen COL3) were calculated using the 2^-ΔΔCt method, with GAPDH as the internal reference gene (primer sequences of GAPDH: F: 5'-ACAACTTTGGTATCGTGGAAGG-3'; R: 5'-GCCATCACGCCACAGTTTC-3').

[0169] 3.1 Anti-aging effect of filtrate (qPCR)

[0170] Table 8 qPCR experimental design table

[0171]

[0172] MMP1 PCR

[0173] Primers for Matrix metallopeptidase 1 (MMP1):

[0174] F:5'-AAAATTACACGCCAGATTTGCC-3'

[0175] R:5'-GGTGTGACATTACTCCAGAGTTG-3'

[0176] MMP1 is a matrix metalloproteinase, which mainly decomposes interstitial collagen, including types I, II and III. Collagen is the main structural protein in the dermis of the skin. When collagen is degraded, the mechanical support of the skin is lost, which leads to the appearance of wrinkles. After the test object acts on the cell model, the anti-wrinkle effect of the test object is evaluated by detecting the changes in MMP1 gene expression.

[0177] Table 9 Relative mRNA levels of MMP1 gene in filtrates of different concentrations

[0178]

[0179] Summary: After UVA treatment, compared with BC, the relative mRNA level of MMP1 gene was significantly upregulated, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP1 gene decreased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, the relative mRNA level of MMP1 gene decreased significantly after 0.08mg / ml and 0.4mg / ml filtrate treatment, while the relative mRNA level of MMP1 gene increased significantly after 0.8mg / ml filtrate treatment. This result shows that the filtrate concentration of 0.08mg / ml and 0.4mg / ml can significantly inhibit the expression of MMP1 gene, and has a certain anti-aging effect.

[0180] MMP3 PCR

[0181] Primers for Matrix metallopeptidase 3 (MMP3):

[0182] F:5'-CTGGACTCCGACACTCTGGA-3'

[0183] R:5'-CAGGAAAGGTTCTGAAGTGACC-3'

[0184] MMP3 is a matrix metalloproteinase III that can degrade some protein components in the extracellular matrix, such as collagen. In the skin, the degradation of collagen is associated with skin aging. Some studies have shown that MMP3 may play a role in tissue remodeling of the skin, helping to improve the elasticity and firmness of the skin by promoting the repair and regeneration of the extracellular matrix. In addition, MMP3 may be involved in regulating the proliferation and migration of skin cells. This is essential for the repair and regeneration process of the skin. After the test substance is applied to the cell model, the anti-wrinkle effect of the test substance is evaluated by detecting changes in MMP3 gene expression.

[0185] Table 10 Relative mRNA levels of MMP3 gene in filtrates of different concentrations

[0186]

[0187] Summary: After UVA treatment, compared with BC, the relative mRNA level of MMP3 gene was significantly upregulated, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP3 gene decreased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, the relative mRNA level of MMP3 gene decreased significantly after treatment with 0.4mg / ml and 0.8mg / ml filtrate. This result shows that the filtrate concentration of 0.4mg / ml and 0.8mg / ml can significantly inhibit the expression of MMP3 gene, and has a certain anti-aging effect.

[0188] COL1 PCR

[0189] Primers for Collagen I (COL1):

[0190] F:5'- GTGCGATGACTGATCTGTGA-3'

[0191] R:5'- CGGTGGTTTCTTGGTCGGT-3'

[0192] COL1 is type I collagen, which accounts for about 80% of the dermis of the skin. Type I collagen helps the dermis (middle layer of the skin) form fibroblasts, helps new cells grow to provide structure, strength and support for the skin, makes the skin plump and promotes the expression of COL1 protein, which can achieve a certain effect in resisting the formation of wrinkles, thereby achieving anti-aging effects.

[0193] Table 11 Relative mRNA level values ​​of COL1 gene in filtrates of different concentrations

[0194]

[0195] Summary: After UVA treatment, compared with BC, the relative mRNA level of COL1 gene decreased significantly, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of COL1 gene increased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, the relative mRNA level of COL1 gene increased significantly after treatment with different concentrations of filtrate. This result shows that the filtrate can significantly increase the expression of COL1 gene in cells and has a certain anti-aging effect.

[0196] COL3 PCR

[0197] Primers for Collagen III (COL3):

[0198] F: 5'-TTGAAGGAGGATGTTCCCATCT-3'

[0199] R: 5'-ACAGACACATATTTGGCATGGTT-3

[0200] COL3 is type III collagen, one of the most abundant proteins in the human body. It is composed of three alpha chains that contain many amino acids, mainly glycine, hydroxyproline and proline. COL3 is mainly found in the skin, blood vessel walls, internal organs and muscles. The functions of COL3 are similar to other types of collagen, including providing strength and elasticity to tissues, maintaining the firmness and elasticity of the skin, supporting the structure of blood vessel walls, and maintaining the health of muscles and joints.

[0201] Table 12 Relative mRNA levels of COL3 gene in filtrates of different concentrations

[0202]

[0203] Summary: After UVA treatment, compared with BC, the relative mRNA level of COL3 gene decreased significantly, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of COL3 gene increased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, the relative mRNA level of COL3 gene increased significantly after treatment with different concentrations of filtrate. This result shows that the filtrate can significantly increase the expression of COL3 gene in cells and has a certain anti-aging effect.

[0204] 3.2 Anti-aging effect of microcurrent tetrapeptide (qPCR)

[0205] Table 13 qPCR experimental design table

[0206]

[0207] MMP1 PCR

[0208] Primers for Matrix metallopeptidase 1 (MMP1):

[0209] F:5'-AAAATTACACGCCAGATTTGCC-3'

[0210] R:5'-GGTGTGACATTACTCCAGAGTTG-3'

[0211] MMP1 is a matrix metalloproteinase, which mainly decomposes interstitial collagen, including types I, II and III. Collagen is the main structural protein in the dermis of the skin. When collagen is degraded, the mechanical support of the skin is lost, which leads to the appearance of wrinkles. After the test object acts on the cell model, the anti-wrinkle effect of the test object is evaluated by detecting the changes in MMP1 gene expression.

[0212] Table 14 Relative mRNA levels of MMP1 gene in different concentrations of microcurrent tetrapeptide

[0213]

[0214] Summary: After UVA treatment, compared with BC, the relative mRNA level of MMP1 gene was significantly upregulated, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP1 gene decreased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, after 0.08mg / ml and 0.4mg / ml microcurrent tetrapeptide treatment, the relative mRNA level of MMP1 gene increased significantly, while after 0.8mg / ml microcurrent tetrapeptide treatment, the relative mRNA level of MMP1 gene decreased significantly. This result shows that the microcurrent tetrapeptide concentration of 0.8mg / ml can significantly inhibit the expression of MMP1 gene and has a certain anti-aging effect.

[0215] MMP3 PCR

[0216] Primers for Matrix metallopeptidase 3 (MMP3):

[0217] F:5'-CTGGACTCCGACACTCTGGA-3'

[0218] R:5'-CAGGAAAGGTTCTGAAGTGACC-3'

[0219] MMP3 is a matrix metalloproteinase III that can degrade some protein components in the extracellular matrix, such as collagen. In the skin, the degradation of collagen is associated with skin aging. Some studies have shown that MMP3 may play a role in tissue remodeling of the skin, helping to improve the elasticity and firmness of the skin by promoting the repair and regeneration of the extracellular matrix. In addition, MMP3 may be involved in regulating the proliferation and migration of skin cells. This is essential for the repair and regeneration process of the skin. After the test substance is applied to the cell model, the anti-wrinkle effect of the test substance is evaluated by detecting changes in MMP3 gene expression.

[0220] Table 15 Relative mRNA levels of MMP3 gene in different concentrations of microcurrent tetrapeptide

[0221]

[0222] Summary: After UVA treatment, compared with BC, the relative mRNA level of MMP3 gene was significantly upregulated, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP3 gene decreased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, after 0.08mg / ml microcurrent tetrapeptide treatment, the relative mRNA level of MMP3 gene increased significantly, while after 0.4mg / ml and 0.8mg / ml microcurrent tetrapeptide treatment, the relative mRNA level of MMP3 gene decreased significantly. This result shows that the microcurrent tetrapeptide concentration of 0.4mg / ml and 0.8mg / ml can significantly inhibit the expression of MMP3 gene, and has a certain anti-aging effect.

[0223] COL1 PCR

[0224] Primers for Collagen I (COL1):

[0225] F:5'- GTGCGATGACTGATCTGTGA-3'

[0226] R:5'- CGGTGGTTTCTTGGTCGGT-3'

[0227] COL1 is type I collagen, which accounts for about 80% of the dermis of the skin. Type I collagen helps the dermis (middle layer of the skin) form fibroblasts, helps new cells grow to provide structure, strength and support for the skin, makes the skin plump and promotes the expression of COL1 protein, which can achieve a certain effect in resisting the formation of wrinkles, thereby achieving anti-aging effects.

[0228] Table 16 Relative mRNA level values ​​of COL1 gene in different concentrations of microcurrent tetrapeptide

[0229]

[0230] Summary: After UVA treatment, compared with BC, the relative mRNA level of COL1 gene decreased significantly, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of COL1 gene increased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, the relative mRNA level of COL1 gene increased significantly after treatment with different concentrations of microcurrent tetrapeptide. This result shows that microcurrent tetrapeptide can significantly increase the expression of COL1 gene in cells and has a certain anti-aging effect.

[0231] COL3 PCR

[0232] Primers for Collagen III (COL3):

[0233] F: 5'-TTGAAGGAGGATGTTCCCATCT-3'

[0234] R: 5'-ACAGACACATATTTGGCATGGTT-3

[0235] COL3 is type III collagen, one of the most abundant proteins in the human body. It is composed of three alpha chains that contain many amino acids, mainly glycine, hydroxyproline and proline. COL3 is mainly found in the skin, blood vessel walls, internal organs and muscles. The functions of COL3 are similar to other types of collagen, including providing strength and elasticity to tissues, maintaining the firmness and elasticity of the skin, supporting the structure of blood vessel walls, and maintaining the health of muscles and joints.

[0236] Table 17 Relative mRNA level values ​​of COL3 gene in different concentrations of microcurrent tetrapeptide

[0237]

[0238] Summary: After UVA treatment, compared with BC, the relative mRNA level of COL3 gene decreased significantly, indicating that the anti-aging model was successfully established. Under UVA stimulation, using the positive control TGF-β, the relative mRNA level of COL3 gene increased significantly, indicating that the positive control of this experiment was effective. Compared with the UVA alone treatment group, after 0.08mg / ml and 0.4mg / ml microcurrent tetrapeptide treatment, the relative mRNA level of COL3 gene increased significantly. This result shows that the microcurrent tetrapeptide concentration has a certain anti-aging effect at 0.08mg / ml and 0.4mg / ml.

[0239] 3.3 Filtrate + microcurrent tetrapeptide anti-aging synergistic effect (qPCR)

[0240] Table 18 qPCR experimental design table

[0241]

[0242] MMP1 PCR

[0243] Primers for Matrix metallopeptidase 1 (MMP1):

[0244] F:5'-AAAATTACACGCCAGATTTGCC-3'

[0245] R:5'-GGTGTGACATTACTCCAGAGTTG-3'

[0246] MMP1 is a matrix metalloproteinase, which mainly decomposes interstitial collagen, including types I, II and III. Collagen is the main structural protein in the dermis of the skin. When collagen is degraded, the mechanical support of the skin is lost, which leads to the appearance of wrinkles. After the test object acts on the cell model, the anti-wrinkle effect of the test object is evaluated by detecting the changes in MMP1 gene expression.

[0247] Table 19 Relative mRNA level values ​​of MMP1 gene in different ratios of filtrate and microcurrent tetrapeptide

[0248] (The dosage concentrations under different ratios are the same as those in Table 18)

[0249]

[0250] Summary: After UVA treatment, the relative mRNA level of MMP1 gene increased significantly compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP1 gene decreased significantly, indicating that the positive control of the experiment was effective. The gene expression level of MMP1 in all filtrates and microcurrent tetrapeptide compositions did not change significantly compared with that of the single microcurrent tetrapeptide, indicating that the filtrate and microcurrent tetrapeptide did not synergistically regulate the expression of the MMP1 gene.

[0251] MMP3 PCR

[0252] Primers for Matrix metallopeptidase 3 (MMP3):

[0253] F:5'-CTGGACTCCGACACTCTGGA-3'

[0254] R:5'-CAGGAAAGGTTCTGAAGTGACC-3'

[0255] MMP3 is a matrix metalloproteinase III that can degrade some protein components in the extracellular matrix, such as collagen. In the skin, the degradation of collagen is associated with skin aging. Some studies have shown that MMP3 may play a role in tissue remodeling of the skin, helping to improve the elasticity and firmness of the skin by promoting the repair and regeneration of the extracellular matrix. In addition, MMP3 may be involved in regulating the proliferation and migration of skin cells. This is essential for the repair and regeneration process of the skin. After the test substance is applied to the cell model, the anti-wrinkle effect of the test substance is evaluated by detecting changes in MMP3 gene expression.

[0256] Table 20 Relative mRNA level values ​​of MMP3 gene in different ratios of filtrate and microcurrent tetrapeptide

[0257] (The dosage concentrations under different ratios are the same as those in Table 18)

[0258]

[0259] Summary: After UVA treatment, the relative mRNA level of MMP3 gene increased significantly compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of MMP1 gene decreased significantly, indicating that the positive control of this experiment was effective. When the ratio of filtrate and microcurrent tetrapeptide combination was 10:1, the gene expression level of MMP1 was significantly decreased compared with that of the single filtrate and microcurrent tetrapeptide, indicating that the filtrate and microcurrent tetrapeptide have synergistic anti-aging effects.

[0260] COL1 PCR

[0261] Primers for Collagen I (COL1):

[0262] F:5'- GTGCGATGACTGATCTGTGA-3'

[0263] R:5'- CGGTGGTTTCTTGGTCGGT-3'

[0264] COL1 is type I collagen, which accounts for about 80% of the dermis of the skin. Type I collagen helps the dermis (middle layer of the skin) form fibroblasts, helps new cells grow to provide structure, strength and support for the skin, makes the skin plump and promotes the expression of COL1 protein, which can achieve a certain effect in resisting the formation of wrinkles, thereby achieving anti-aging effects.

[0265] Table 21 Relative mRNA level values ​​of COL1 gene in different ratios of filtrate and microcurrent tetrapeptide

[0266] (The dosage concentrations under different ratios are the same as those in Table 18)

[0267]

[0268] Summary: After UVA treatment, the relative mRNA level of COL1 gene decreased significantly compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of COL1 gene increased significantly, indicating that the positive control of this experiment was effective. When the ratio of filtrate and microcurrent tetrapeptide combination was 10:1, the gene expression level of COL1 increased significantly compared with the single filtrate and microcurrent tetrapeptide, and S BLISS >0, indicating that the filtrate and microcurrent tetrapeptide have a synergistic anti-aging effect, indicating that the filtrate and microcurrent tetrapeptide combination have a synergistic effect in anti-aging efficacy.

[0269] COL3 PCR

[0270] Primers for Collagen III (COL3):

[0271] F: 5'-TTGAAGGAGGATGTTCCCATCT-3'

[0272] R: 5'-ACAGACACATATTTGGCATGGTT-3

[0273] COL3 is type III collagen, one of the most abundant proteins in the human body. It is composed of three alpha chains that contain many amino acids, mainly glycine, hydroxyproline and proline. COL3 is mainly found in the skin, blood vessel walls, internal organs and muscles. The functions of COL3 are similar to other types of collagen, including providing strength and elasticity to tissues, maintaining the firmness and elasticity of the skin, supporting the structure of blood vessel walls, and maintaining the health of muscles and joints.

[0274] Table 22 Relative mRNA level values ​​of COL3 gene in different ratios of filtrate and microcurrent tetrapeptide

[0275] (The dosage concentrations under different ratios are the same as those in Table 18)

[0276]

[0277] Summary: After UVA treatment, the relative mRNA level of COL3 gene decreased significantly compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the relative mRNA level of COL3 gene increased significantly, indicating that the positive control of this experiment was effective. When the ratio of filtrate and microcurrent tetrapeptide combination was 10:1, the gene expression level of COL3 increased significantly compared with the single filtrate and microcurrent tetrapeptide, and S BLISS >0, indicating that the filtrate and microcurrent tetrapeptide have a synergistic anti-aging effect, indicating that the filtrate and microcurrent tetrapeptide combination have a synergistic effect in anti-aging efficacy.

[0278] Detection and analysis of four anti-aging proteins (WB)

[0279] (1) HFF fibroblasts grown in 12-well plates were stimulated with UVA to establish the model.

[0280] (2) 24 hours after modeling, cells were collected and extracted by adding RIPA cell lysis buffer.

[0281] (3) Protein quantification: Use a protein quantification method (such as the BCA method) to determine the extracted protein concentration.

[0282] (4) SDS-PAGE electrophoresis: Use polyacrylamide gel electrophoresis (SDS-PAGE) to separate protein samples according to their molecular weight.

[0283] (5) Transfer: Assemble the gel, membrane and transfer buffer in a transfer apparatus, and use a wet transfer apparatus to transfer the separated proteins from the gel to the PVDF membrane.

[0284] (6) Blocking: Place the membrane in 5% skimmed milk powder and incubate for 1 h.

[0285] (7) Primary antibody incubation: Add the primary antibody (specific antibody) of the target protein to 5% skim milk powder and incubate overnight to allow it to fully bind to the target protein in the membrane.

[0286] (8) Washing: Wash the membrane with buffer TBST to remove unbound primary antibody.

[0287] (9) Secondary antibody incubation: Add the secondary antibody corresponding to the source of the primary antibody (e.g., goat anti-rabbit IgG) to a solution containing 5% skim milk powder and incubate for 2 h to allow it to bind to the primary antibody. The secondary antibody is usually labeled with an enzyme (e.g., horseradish peroxidase-HRP) or a fluorescent dye.

[0288] (10) Washing: Wash the membrane with buffer TBST to remove unbound secondary antibody.

[0289] (11) Signal detection: The secondary antibody-labeled protein was detected using the substrate chemiluminescence ECL method, and the target protein was imaged and stored using the ChemiDoc imaging system.

[0290] (12) Result analysis: ImageJ was used to quantitatively analyze the target protein based on the signal intensity and location.

[0291] 4.1 Anti-aging effect of filtrate + microcurrent tetrapeptide (10:1) (WB)

[0292] Table 23 WB experimental design table

[0293]

[0294] MMP1 WB

[0295] MMP1 is a matrix metalloproteinase, which mainly decomposes interstitial collagen, including types I, II and III. Collagen is the main structural protein in the dermis of the skin. When collagen is degraded, the mechanical support of the skin is lost, which leads to the appearance of wrinkles. After the test object acts on the cell model, the anti-wrinkle effect of the test object is evaluated by detecting the changes in the expression of MMP1 protein.

[0296] Table 24 Relative expression values ​​of MMP1 protein in filtrate, microcurrent tetrapeptide and different proportions of the composition

[0297] (The dosage concentrations at different ratios are the same as those in Table 23)

[0298]

[0299] Summary: After UVA treatment, the expression of MMP1 protein increased compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the expression of MMP1 protein decreased significantly using the positive control TGF-β, indicating that the positive control of this experiment was effective. Under UVA stimulation, the expression of MMP1 protein in all monomers and compositions decreased significantly, indicating that all monomers and compositions have anti-aging effects. In addition, when the ratio of the filtrate and microcurrent tetrapeptide composition was 10:1, the expression level of MMP1 protein decreased significantly compared with the single filtrate and microcurrent tetrapeptide, and S BLISS >0, indicating that the filtrate and microcurrent tetrapeptide have a synergistic anti-aging effect. At the same time, compared with the combination of CLR bifid yeast and microcurrent tetrapeptide (10:1), the MMP1 protein expression level of the filtrate and microcurrent tetrapeptide combination (10:1) showed a significant decrease, indicating that the combination of filtrate and microcurrent tetrapeptide has a higher inhibition rate on MMP1 protein than the combination of CLR bifid yeast and microcurrent tetrapeptide. In summary, the combination of filtrate and microcurrent tetrapeptide has a synergistic effect on anti-aging efficacy, and the effect is better than the anti-aging effect of the combination of CLR bifid yeast and microcurrent tetrapeptide.

[0300] COL1 WB

[0301] COL1 is type I collagen, which accounts for about 80% of the dermis of the skin. Type I collagen helps the dermis (middle layer of the skin) form fibroblasts, helps new cells grow to provide structure, strength and support for the skin, makes the skin plump and promotes the expression of COL1 protein, which can achieve a certain effect in resisting the formation of wrinkles, thereby achieving anti-aging effects.

[0302] Table 25 Relative expression values ​​of COL1 protein in filtrate, microcurrent tetrapeptide and different proportions of the composition

[0303] (The dosage concentrations at different ratios are the same as those in Table 23)

[0304]

[0305] Summary: After UVA treatment, the expression of COL1 protein decreased significantly compared with BC, indicating that the anti-aging model was successfully established. Under UVA stimulation, the positive control TGF-β was used, and the expression of COL1 protein increased significantly, indicating that the positive control of this experiment was effective. Under UVA stimulation, the expression of COL1 protein in 0.8 mg / ml filtrate increased significantly, indicating that the filtrate has anti-aging effects. In addition, when the ratio of the filtrate and microcurrent tetrapeptide combination was 10:1, the expression level of COL1 protein was significantly increased compared with the single filtrate and microcurrent tetrapeptide, and S BLISS >0, indicating that the filtrate and microcurrent tetrapeptide have a synergistic anti-aging effect. At the same time, compared with the combination of CLR bifid yeast and microcurrent tetrapeptide (10:1), the COL1 protein expression level of the filtrate and microcurrent tetrapeptide combination (10:1) showed a significant increase, indicating that the combination of filtrate and microcurrent tetrapeptide significantly increased the expression of COL1 protein compared with the combination of CLR bifid yeast and microcurrent tetrapeptide. In summary, the combination of filtrate and microcurrent tetrapeptide has a synergistic effect on anti-aging efficacy, and the effect is better than the anti-aging effect of the combination of CLR bifid yeast and microcurrent tetrapeptide.

[0306] 5. Antibacterial detection and analysis

[0307] (1) Take out the glycerol storage tube of the strain from the -80°C ultra-low temperature freezer, streak a single colony on the prepared R2A solid plate, and then place it in a 30°C incubator for 48 h;

[0308] (2) Take the R2A culture plate out of the 30°C incubator and pick a single colony with an inoculation loop in a clean bench. Place the colonies in sterile 12 ml shaking tubes and add 4 ml of R2A liquid culture medium to each tube. Place the tubes in a constant temperature shaker at 220 rpm and culture overnight at 30°C.

[0309] (3) Take the bacterial solution in step 2, prepare a sterile cell shake flask, take a certain volume, inoculate it into fresh sterile R2A liquid culture medium at a ratio of 2%, and then place it in a constant temperature shaker at a speed of 220 rpm and 30°C for 6 h;

[0310] (4) Meanwhile, prepare several sterile 96-well plates, add 30ul [ddH2O / (0.5% v / v) DMSO] to the negative control group, add 30ul R2A liquid culture medium and other components to the positive control group, and add 30ul raw materials to the experimental group, and set aside for use;

[0311] (5) Take 200ul of the bacterial solution from step 3, add it to a sterile 96-well plate, and measure the OD600; then resuspend it in fresh sterile R2A medium and adjust its OD600 reading to between 0.14 and 0.16;

[0312] (6) Mix the resuspended bacterial solution and biolog dye in the ratio (2ul: 170ul = 1: 85) (protect from light), and add 172ul to a 96-well plate;

[0313] (7) Measure the OD590 of each 96-well plate at 0 h, and then place it in a 30°C incubator in the dark;

[0314] (8) Measure OD590 at different time periods and compare and record the data.

[0315] 5.1 Filtrate + microcurrent tetrapeptide (10:1) antibacterial experiment

[0316] Table 26 Antibacterial experiment design table

[0317]

[0318] 5.2 Moraxella osloensis

[0319] Moraxella osloensis is a normal flora of human skin, mucous membranes and respiratory tract. Previous studies have shown that Moraxella osloensis is the second most abundant species in Chinese skin samples and can be used as an indicator of aging in Han Chinese. Moraxella osloensis is involved in regulating collagen metabolism, extracellular matrix assembly and promoting cell aging in human keratinocytes and fibroblasts.

[0320] Table 27 Numerical table of relative growth rates of bacterial flora in filtrate, microcurrent tetrapeptide and combination

[0321] (The dosage concentrations at different ratios are the same as those in Table 26)

[0322]

[0323] Summary: After culturing Moraxella osloensis for 24 hours, the growth rate of the bacterial colony after R2A treatment was significantly increased compared with BC, while the growth rate of the bacterial colony after treatment with the filtrate and the microcurrent tetrapeptide composition was significantly decreased compared with BC. This shows that the composition can inhibit the growth of Moraxella osloensis to a certain extent. After culturing Moraxella osloensis for 48 hours and 72 hours, the growth rate of the bacterial colony after treatment with the filtrate, microcurrent tetrapeptide and their combination was significantly decreased compared with BC. This shows that the filtrate and microcurrent tetrapeptide can significantly inhibit the growth of Moraxella osloensis and have certain microecological anti-aging effects. In addition, when the ratio of the filtrate and the microcurrent tetrapeptide composition was 10:1, the growth rate of the bacterial colony was significantly decreased compared with the filtrate and microcurrent tetrapeptide alone, and S BLISS >0, indicating that the filtrate and microcurrent tetrapeptide can synergistically inhibit the growth of Moraxella osloensis and have good microecological anti-aging effects.

[0324] 6. Synergy Calculation

[0325] The above data were collected and the Bliss synergy score was calculated according to the Bliss independence model. The formula is S BLISS =E A,B -(E A +E B -E A E B ), where S BLISS <0 is antagonistic, S BLISS =0 is additive effect, S BLISS >0 indicates synergistic effect. The E value is the average value of the test sample effect minus the average value of the control group after corresponding UVA treatment. The results are shown in Table 28:

[0326] Table 28

[0327]

[0328] Note: The ratio refers to the mass ratio of the filtrate and the microcurrent tetrapeptide.

[0329] The above results show that in most cases, the two components in the experimental composition have a good synergistic effect. Therefore, the combination of these two components can have a good synergistic anti-aging effect.

[0330] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition comprising a fermentation filtrate of Bifidobacterium longum subsp. infantis and a microcurrent tetrapeptide; in, The mass ratio of the fermentation filtrate of Bifidobacterium longum subsp. infantis to the microcurrent tetrapeptide is 1:10 to 10:1; The preparation method of the fermentation filtrate of Bifidobacterium longum subspecies infantis comprises: inoculating and culturing Bifidobacterium longum subspecies infantis with a preservation number of CGMCC NO.27851 to obtain a seed solution; fermenting the seed solution, centrifuging it, and taking the supernatant to obtain the fermentation filtrate of Bifidobacterium longum subspecies infantis; The peptide sequence of the microcurrent tetrapeptide is LEAP.

2. The composition according to claim 1, characterized in that The mass ratio of the fermentation filtrate of Bifidobacterium longum subspecies infantis to the microcurrent tetrapeptide is 1:10, 1:1 or 10:

1.

3. Use of the composition according to claim 1 or 2 in the preparation of anti-aging products.

4. The use according to claim 3, characterized in that: The anti-aging includes anti-photodamage and / or inhibition of harmful bacteria.

5. The use according to claim 4, characterized in that: The anti-light damage includes regulating the expression level of related proteins, regulating the transcription level of related genes and / or improving the ability of cells to resist ultraviolet damage; The harmful bacteria is Moraxella osloensis.

6. The use according to claim 5, characterized in that: The related proteins are type I collagen and / or matrix metalloproteinases; The related genes are COL3, COL 1, MMP3 and / or MMP1 ; The regulation includes increasing COL 1 Genes and / or COL3 The transcription level of the gene is reduced MMP1 Genes and / or MMP3 Transcription levels of genes; The cells are fibroblasts; The ultraviolet damage is the damage caused by UVA.

7. An anti-aging product comprising the composition according to claim 1 or 2.

8. The product according to claim 7, characterized in that in, The concentration of the fermentation filtrate of Bifidobacterium longum subspecies infantis is 0.08 mg / ml-20 mg / ml, or its mass fraction is 0.008%-2%, and the concentration of the microcurrent tetrapeptide is 0.08 mg / ml-20 mg / ml, or its mass fraction is 0.008%-2%.

9. The product according to claim 7 or 8, characterized in that It is a cosmetic, which also includes cosmetically acceptable auxiliary materials.

10. The product according to claim 9, characterized in that The cosmetics are cleaning cosmetics, care cosmetics or beauty / modification cosmetics, and their dosage forms include creams, emulsions, solutions, sprays, gels or powders.

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