Application of active peptide in preparation of product for improving skin barrier and promoting collagen expression

Abstract

The invention provides application of an active peptide in preparation of a product for improving skin barrier and promoting collagen expression, the structural formula of the active peptide is H-A-OH, A comprises at least two amino acid residues, and the amino acid residues comprise at least one of Hyp (hydroxyproline), Gly (glycine), Pro (proline) and Ala (alanine). The invention proves that the collagen expression and skin state of the organism can be improved by supplementing exogenous precursors such as active peptides, and the polypeptide has wide application prospects in the fields of dietary supplements, pet foods and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of common food, health food, pet food or medicine, and specifically relates to the use of active peptides in preparing products that improve skin barrier and promote collagen expression. Background Art

[0002] Skin is the body's largest organ, and the most direct manifestation of aging is skin aging. The extracellular matrix (ECM) is a complex structure composed of biomacromolecules produced and secreted by cells onto their surfaces or between cells. It plays a vital role in maintaining cell function and tissue integrity. Composed of various proteins such as collagen, glycoproteins, and proteoglycans, the ECM provides mechanical support, stores growth factors, and regulates cellular homeostasis.

[0003] Collagen not only maintains the morphological structure of tissues but also plays a vital role in various physiological and pathological processes. Its absence or mutation can have serious consequences for the body. Over 1,300 human diseases are known to be caused by collagen mutations. Currently, a growing number of reports demonstrate the relationship between collagen and the immune system.

[0004] Many natural active ingredients play an anti-skin aging role by improving the growth and metabolism of human skin cells, promoting the increase of cell vitality and the expression of skin matrix. Peptides have become the main ingredients of anti-skin aging due to their significant anti-skin aging effects. However, in the current existing technologies, most of them extract collagen peptides from animal fish skin, pig skin, cowhide and tendon, and then use them to prepare related products for improving skin aging or skin condition. However, in these existing technologies, the actual components of the collagen small molecule peptide protein that play a role are not explained, nor are the effects of the intervention explained from the perspective of natural aging. Summary of the Invention

[0005] To address the problems and deficiencies in the prior art, the present invention provides a method for preparing a product that improves the skin barrier and promotes collagen expression. The present invention demonstrates that supplementing exogenous precursors such as active peptides can improve the body's collagen expression and skin condition, and has broad application prospects in multiple fields such as dietary supplements and pet food. Furthermore, in the use of the active peptides provided by the present invention for preparing products that improve the skin barrier and promote collagen expression, the related products include at least one of ordinary food, health food, pet food, and medicine, which can meet the needs of various market sectors.

[0006] According to a first aspect of the present invention, there is provided a use of an active peptide for preparing a product for improving the skin barrier and promoting collagen expression. The active peptide has a structural formula of HA-OH, where A comprises at least two amino acid residues, and the amino acid residues include at least one of Hyp (hydroxyproline), Gly (glycine), Pro (proline), and Ala (alanine).

[0007] As the largest organ in the human body, the skin is the first to experience the changes of aging. Skin aging leads to thinning of the epidermis and dermis, and damage to the skin barrier (physical barrier, immune barrier, etc.). The basement membrane is the key connecting structure between the dermis and epidermis. The health of the basement membrane is crucial for normal skin metabolism and barrier function. During skin aging, the expression of basement membrane components, such as collagen XVII, decreases. Supplementing extracellular matrix components such as active peptides can enhance the regeneration, differentiation, and migration of basement membrane cells, which is crucial for repairing the skin barrier.

[0008] Active peptide components are precursors to collagen in the human body. This study demonstrates that supplementing with exogenous precursors can improve collagen expression and skin condition. Specifically, the active peptides have been shown to effectively promote the expression of collagens such as COL12 and COL19, and reduce skin barrier damage, specifically by reducing NLP-29 expression. Therefore, using these active peptides can effectively improve the condition of naturally aging skin, resulting in smoother, more translucent, and healthier skin.

[0009] In addition, the confirmation experiment of the present invention used Caenorhabditis elegans as a model organism to verify the effectiveness of the above-mentioned active peptides. Collagen is the most important component of ECM, and the extracellular matrix of nematodes is mainly distributed in the stratum corneum and basement membrane. Caenorhabditis elegans is a classic model organism for studying aging. It has a short life cycle and a clear genetic background. Using Caenorhabditis elegans as a model organism to study the effects of natural aging on the skin barrier has certain innovations. Caenorhabditis elegans also has the advantages of a clear genetic background, a simple individual structure, a short life history, complete genome sequencing, and no ethical issues. It has a wide range of applications in the fields of genetics and developmental biology, aging and lifespan. The skin barrier is considered to be an important indicator of aging, and the epidermis of Caenorhabditis elegans has a high homology with human skin. Therefore, Caenorhabditis elegans was selected as the model organism for our functional verification.

[0010] Furthermore, the present invention uses the naturally aging Caenorhabditis elegans nematode as a model organism. This is the first demonstration, utilizing the naturally aging skin barrier model of the nematode, that the aforementioned active peptides improve the skin barrier and other skin conditions, such as epidermal wrinkles, under naturally aging conditions. This provides a new strategy for the clinical prevention of impaired skin barrier in aging, and has promising application prospects. Furthermore, this invention is the first to use the aforementioned active peptides to improve the skin barrier impaired under naturally aging conditions, expanding the application of products containing the corresponding components in naturally aging populations. Furthermore, this invention is the first to utilize the naturally aging nematode in skin-related intervention research, expanding the application of nematodes in skin beauty.

[0011] Preferably, products that improve the skin barrier and promote collagen expression include at least one of H-Hyp-Gly-OH (OG), H-Gly-Pro-Ala-OH (GPA), H-Gly-Pro-Hyp-OH (GPO), and H-Gly-Pro-Hyp-Gly-OH (GPOG). These collagen peptides, acting alone or in combination, have been shown in relevant experiments to effectively improve the skin barrier and promote collagen expression. This means they can effectively improve skin condition and delay skin aging during natural aging, demonstrating promising prospects for intervention in natural aging. It should be noted here that, with respect to H-Hyp-Gly-OH (OG), Hyp-Gly corresponds to sequence 1 in the sequence listing, i.e., residue "XG"; with respect to H-Gly-Pro-Ala-OH (GPA), Gly-Pro-Ala corresponds to sequence 2 in the sequence listing, i.e., residue "GPA"; with respect to H-Gly-Pro-Hyp-OH (GPO), Gly-Pro-Hyp corresponds to sequence 3 in the sequence listing, i.e., residue "GPX"; with respect to H-Gly-Pro-Hyp-Gly-OH (GPOG), Gly-Pro-Hyp-Gly corresponds to sequence 4 in the sequence listing, i.e., residue "GPXG".

[0012] Preferably, promoting collagen expression includes at least one of the following: (1) promoting the expression of collagen COL-12; (2) promoting the expression of collagen COL-19; (3) promoting the expression of collagen EMB-9; (4) improving epidermal wrinkles; and improving the skin barrier includes reducing the expression of the antimicrobial protein peptide NLP-29. Relevant experiments have verified that the present invention has verified that the above-mentioned active peptides can promote the expression of collagens COL-12, COL-19, and EMB-9 and reduce the expression of the antimicrobial protein peptide NLP-29, thereby effectively improving the damaged skin barrier and wrinkles caused by natural aging and improving skin condition.

[0013] Preferably, the product for improving skin barrier and promoting collagen expression is applicable to animals and humans.

[0014] Preferably, the product that improves the skin barrier and promotes collagen expression is used to improve the damaged skin barrier of animals or humans in natural aging, and to promote collagen expression in humans or animals in natural aging.

[0015] Preferably, improving the skin barrier and promoting collagen expression includes: H-Hyp-Gly-OH (OG), and / or H-Gly-Pro-Ala-OH (GPA), and / or H-Gly-Pro-Hyp-OH (GPO), and / or H-Gly-Pro-Hyp-Gly-OH (GPOG) are used to increase the expression of collagen COL-12 in middle-aged animals or humans.

[0016] Preferably, improving the skin barrier and promoting collagen expression includes: H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-12 in elderly animals or humans.

[0017] Preferably, improving the skin barrier and promoting collagen expression includes: H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-19 in young animals or humans.

[0018] Preferably, improving the skin barrier and promoting collagen expression includes: H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-19 in middle-aged animals or humans.

[0019] Preferably, improving the skin barrier and promoting collagen expression includes: H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-19 in elderly animals or humans.

[0020] Preferably, improving the skin barrier and promoting collagen expression includes: H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to reduce the expression of antimicrobial protein peptide NLP-29 in middle-aged animals or humans.

[0021] Preferably, improving the skin barrier and promoting collagen expression include: H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH for improving epidermal wrinkles in animals or humans in middle age.

[0022] Preferably, improving the skin barrier and promoting collagen expression include: H-Hyp-Gly-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to improve epidermal wrinkles in animals or humans during old age.

[0023] Preferably, improving the skin barrier and promoting collagen expression comprises: H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of basement membrane collagen EMB-9 in young animals or humans.

[0024] Preferably, improving the skin barrier and promoting collagen expression comprises: H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of basement membrane collagen EMB-9 in middle-aged animals or humans.

[0025] Preferably, improving the skin barrier and promoting collagen expression includes: H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH are used to increase the expression of basement membrane collagen EMB-9 in elderly animals or humans.

[0026] Preferably, the product for improving the skin barrier and promoting collagen expression includes an oral preparation, and the oral preparation includes at least one of tablets, capsules, granules, ointments, liquids, pills, and suspensions.

[0027] Preferably, the product that improves the skin barrier and promotes collagen expression includes at least two of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH; or, the product includes at least three of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH; or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH.

[0028] Preferably, the product that improves the skin barrier and promotes collagen expression includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH; or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH; or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH; or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH; or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH, and N-acetylglucosamine.

[0029] Preferably, when the product for improving the skin barrier and promoting the expression of collagen includes H-Hyp-Gly-OH and H-Gly-Pro-Ala-OH, the mass ratio of H-Hyp-Gly-OH and H-Gly-Pro-Ala-OH is 4-6:4-6; or, when the product for improving the skin barrier and promoting the expression of collagen includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-OH, the mass ratio of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-OH is 0.1-3:4-9.8:0.1-3; or, when the product for improving the skin barrier and promoting the expression of collagen includes H-Hyp-Gly-OH, H-Gly-Pro-Hyp- or, when the product for improving skin barrier and promoting collagen expression includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH, the mass ratio of H-Hyp-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH is 0.1~3:4~9.8:0.1~3; or, when the product for improving skin barrier and promoting collagen expression includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH, the mass ratio of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH is 0.1~2:2~3.9:4~5.9:0.1~2.

[0030] According to a second aspect of the present invention, there is provided a use of an active peptide in preparing a common food, a health food, a pet food or a medicine that improves the skin barrier and promotes collagen expression.

[0031] According to a third aspect of the present invention, there is provided a use of an active peptide in preparing a powder, soft candy or beverage for improving the skin barrier and promoting collagen expression.

[0032] According to a fourth aspect of the present invention, there is provided a use of an active peptide and amino sugar in preparing a product for improving the skin barrier and promoting collagen expression.

[0033] Preferably, the active peptide has the structural formula HA-OH, where A comprises at least two amino acid residues, including at least one of Hyp, Gly, Pro, and Ala; and the amino sugar comprises N-acetylglucosamine. The combination of the active peptide and amino sugar can also promote the expression of collagens COL-12, COL-19, and EMB-9, while reducing the expression of the antimicrobial protein peptide NLP-29. This can also effectively improve the damaged skin barrier and wrinkles associated with natural aging.

[0034] Preferably, active peptides and amino sugars are used in the preparation of products that improve the skin barrier and promote collagen expression, and the products include H-Hyp-Gly-OH (OG), H-Gly-Pro-Ala-OH (GPA), H-Gly-Pro-Hyp-OH (GPO), H-Gly-Pro-Hyp-Gly-OH (GPOG), and N-acetylglucosamine (NAG).

[0035] Preferably, the mass ratio of Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH, and N-acetylglucosamine, H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH, and N-acetylglucosamine is 0.2~0.7:3~3.5:4.6~5.2:0.2~0.7:0.3~2.

[0036] In an alternative embodiment, N-acetyl glucosamine is used to prepare a product for improving skin barrier and promoting collagen expression, and improving skin barrier and promoting collagen expression comprises: N-acetyl glucosamine is used to increase the expression of collagen COL-12 in middle-aged animals or humans; In an optional embodiment, N-acetylglucosamine is used to prepare a product that improves the skin barrier and promotes collagen expression, and improving the skin barrier and promoting collagen expression includes: N-acetylglucosamine is used to increase the expression of collagen COL-19 in young animals or humans.

[0037] In an optional embodiment, N-acetylglucosamine is used to prepare a product that improves the skin barrier and promotes collagen expression, and improving the skin barrier and promoting collagen expression includes: N-acetylglucosamine is used to reduce the expression of the antimicrobial protein peptide NLP-29 in young animals or humans.

[0038] In an optional embodiment, N-acetyl glucosamine is used to prepare a product that improves the skin barrier and promotes collagen expression, and improving the skin barrier and promoting collagen expression includes: N-acetyl glucosamine is used to improve epidermal wrinkles in animals or humans in middle age.

[0039] In an optional embodiment, N-acetyl glucosamine is used to prepare a product that improves the skin barrier and promotes collagen expression, and improving the skin barrier and promoting collagen expression includes: N-acetyl glucosamine is used to improve epidermal wrinkles in animals or humans in old age.

[0040] According to a fifth aspect of the present invention, there is provided a use of an active peptide and amino sugar in preparing a common food, health food, pet food or medicine that improves the skin barrier and promotes collagen expression.

[0041] According to a sixth aspect of the present invention, there is provided a use of an active peptide and amino sugar in preparing a powder, soft candy or beverage for improving the skin barrier and promoting collagen expression. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 、 2 These are the expressions of type I collagen COL-12 in the epidermis of nematodes (day 7) and nematodes (day 15) in Example 1.

[0043] Figure 3 、 4 , 5 respectively represent the expression of type II collagen COL-19 in the epidermis of nematodes (day 2 worm age), nematodes (day 7 worm age), and nematodes (day 15 worm age) in Example 2.

[0044] Figure 6 、 7 These are the expression levels of NLP-29 in nematodes (day 2) and nematodes (day 7) in Example 3, respectively.

[0045] Figure 8 、 9 They are SEM images of the epidermis of nematodes (day 7) and nematodes (day 15) in Example 4.

[0046] Figure 10 、 111 and 2 respectively represent the expression of EMB-9 in nematodes (day 2) and nematodes (day 15) in Example 5. DETAILED DESCRIPTION

[0047] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0048] COL-12 is a collagen protein located in the middle layer of the nematode's cuticle. Its expression decreases dramatically in early adulthood, primarily serving a mechanical support role. COL-19 is a collagen protein located in the annular rings and lateral surfaces of the nematode's cuticle. It is expressed in the outermost layer of the epidermis. Its mRNA expression decreases dramatically in early adulthood, but its protein level remains constant. The epidermal cell layer, as a physical barrier, frequently undergoes various physical insults that disrupt its structure. The epidermal cell immune system interprets this structural damage as a danger signal, triggering an immune response and the release of the antimicrobial peptide NLP-29. Increased NLP-29 expression indicates damage to the skin's physical barrier. Day 2 in nematodes is equivalent to 25-30 years old (youth) in humans, Day 7 to 40-50 years old (middle-aged), and Day 15 to 60-70 years old (elderly).

[0049] Therefore, in a specific embodiment of the present invention, active peptides (OG, GPA, GPO, GPOG) and N-acetylglucosamine (NAG) are used to study the expression of COL-12, COL-19, and NLP-29 in nematodes at different stages to verify the effects of the above-mentioned active peptides and N-acetylglucosamine (NAG) on improving the skin barrier and promoting collagen expression during the natural aging process of animals or humans.

[0050] Example 1 The effects of OG, GPA, GPO, GPOG and NAG on the expression of COL-12 were investigated respectively.

[0051] OG, GPA, GPO, GPOG, and NAG were dissolved in water to prepare 1 M solutions, which were then diluted with OP50 (nutrient-deficient Escherichia coli) to prepare 100 mM sample solutions. C. elegans COL-12::DsRed was used as a red fluorescent marker for mid-layer collagen. The amount of mid-layer collagen was determined by the intensity of red fluorescence (higher amounts indicate stronger red fluorescence). Synchronization was performed and, after reaching the L4 stage (48-60 hours), the worms were transferred to FUdR medium (to inhibit egg laying). This was marked as day 0 and cultured at 20°C. Samples were fed on days 0, 5, and 13. Fluorescence imaging and counting of the worms were performed using an upright fluorescence microscope on days 2, 7, and 15, respectively. A minimum of 15 worms were counted each time.

[0052] Fluorescence imaging and intensity statistics results are shown in Figure 2. Figure 1 、 2 As shown, Figure 1 、 2 The expression of type I collagen COL-12 in the epidermis of C. elegans (day 7) and C. elegans (day 15). The mock group was a blank control group, without the addition of active substances OG, GPA, GPO, GPOG, or NAG.

[0053] Depend on Figure 1 It can be seen that compared with the Mock group, the OG group, GPA group, GPO group, GPOG group and NAG group can promote the expression of collagen COL-12 in middle-aged nematodes, with a very significant improvement effect.

[0054] Depend on Figure 2 It can be seen that compared with the Mock group, the GPO group and the GPOG group can promote the expression of collagen COL-12 in elderly nematodes, with a significant improvement effect.

[0055] Example 2 The effects of OG, GPA, GPO, GPOG and NAG on the expression of COL-19 were investigated respectively.

[0056] OG, GPA, GPO, GPOG, and NAG were dissolved in water to prepare 1 M solutions, which were then diluted with OP50 (nutrient-deficient Escherichia coli) to prepare 100 mM sample solutions. Nematodes were synchronized using COL-19::GFP, a green fluorescent marker for mid-layer collagen. The intensity of green fluorescence was used to determine mid-layer collagen content (higher collagen content indicates stronger green fluorescence). After 48-60 hours of growth, nematodes were transferred to FUdR medium (to inhibit egg laying) (day 0) and cultured at 20°C. Samples were fed on days 0, 5, and 13. Fluorescence imaging and counting of nematodes were performed using an upright fluorescence microscope on days 2, 7, and 15, respectively. A minimum of 15 nematodes were counted each time.

[0057] Fluorescence imaging and intensity statistics results are shown in Figure 2. Figure 3 、 4 , 5, where Figure 3 、 4 5 are the expressions of type II collagen COL-19 in the epidermis of nematodes (day 2), nematodes (day 7), and nematodes (day 15), respectively. The mock group is a blank control group, i.e., no active substances OG, GPA, GPO, GPOG, or NAG were added.

[0058] Depend on Figure 3 It can be seen that compared with the Mock group, the OG group, GPA group, GPO group, GPOG group and NAG group can promote the expression of collagen COL-19 in young nematodes, with a very significant improvement effect.

[0059] Depend on Figure 4 It can be seen that compared with the Mock group, the OG group, GPA group, GPO group, and GPOG group can promote the expression of collagen COL-19 in middle-aged nematodes, with a significant improvement effect.

[0060] Depend on Figure 5 It can be seen that compared with the Mock group, the GPO group and the GPOG group can promote the expression of collagen COL-19 in elderly nematodes, with a significant improvement effect.

[0061] Example 3 The effects of OG, GPA, GPO, GPOG and NAG on the expression of NLP-19 were investigated respectively.

[0062] The antimicrobial protein peptide NLP-29 is an immune response product expressed after nematode epidermal damage. The more severe the epidermal damage, the more it is expressed. Therefore, it can be used to measure the extent of epidermal damage.

[0063] OG, GPA, GPO, GPOG, and NAG were dissolved in water to prepare 1 M solutions, which were then diluted with OP50 (nutrient-deficient Escherichia coli) to prepare 100 mM sample solutions. Nematodes were synchronized using NLP-19::GFP, a green fluorescent marker for the middle layer collagen. Green fluorescence intensity indicated damage to the middle layer collagen (lower damage, lower green fluorescence intensity). After 48-60 hours of growth, nematodes were transferred to FUdR medium (to inhibit egg laying) (day 0) and cultured at 20°C. The nematodes were fed samples on days 0 and 5, and were imaged and counted using an upright fluorescence microscope on days 2 and 7, respectively. A minimum of 15 nematodes were counted each time.

[0064] Fluorescence imaging and intensity statistics results are shown in Figure 2. Figure 6 、 7 , 8, where Figure 6 is the expression of NLP-29 in nematodes (day 2), and Figure 6 There are only Mock group and NAG group. Figure 7 The expression of NLP-29 in nematodes (day 7). Figure 8 The expression of NLP-29 in C. elegans (day 15). The Mock group was a blank control group, i.e., no active substances OG, GPA, GPO, GPOG, or NAG were added.

[0065] Depend on Figure 6 It can be seen that compared with the Mock group, the NAG group has a significant protective effect on the epidermis of young nematodes.

[0066] Depend on Figure 7 It can be seen that compared with the Mock group, the OG group, GPA group, GPO group, and GPOG group all have obvious protective effects on the epidermis of middle-aged nematodes.

[0067] Example 4 The effects of OG, GPA, GPO, GPOG and NAG on the expression of epidermal status were investigated respectively.

[0068] OG, GPA, GPO, GPOG, and NAG were dissolved in water to prepare 1 M solutions. These solutions were then diluted with OP50 (nutrient-deficient Escherichia coli) to prepare 100 mM sample solutions. Standard nematodes (N2) were synchronized and grown to the L4 stage (48-60 hours). The nematodes were then transferred to FUdR medium (to inhibit egg laying). This was marked as day 0. The cells were incubated at 20°C with continuous feeding. Scanning electron microscopy (SEM) was used to observe the nematodes on days 7 and 15, with at least five nematodes counted for each image. SEM allows for intuitive visualization of the nematode's epidermis.

[0069] The SEM image results are as follows Figure 8 、 9 As shown, Figure 8 、 9 The following are SEM images of the epidermis of nematodes (day 7) and nematodes (day 15). The mock group is a blank control group, without the addition of active substances OG, GPA, GPO, GPOG, or NAG.

[0070] Depend on Figure 8 It can be seen that the epidermis of middle-aged nematodes in the OG group, GPA group, GPO group, GPOG group and NAG group has a very significant improvement effect.

[0071] Depend on Figure 9 It can be seen that the epidermis of nematodes in the aged period of the OG group, GPO group, GPOG group and NAG group had a very significant improvement effect.

[0072] Example 5 The effects of OG, GPA, GPO, GPOG and NAG on the expression of basement membrane collagen EMB-9 were investigated respectively.

[0073] EMB-9 collagen is a structural component of the extracellular matrix that provides tensile strength. It is involved in multiple processes, including positive regulation of axonal regeneration, protein expression, and serotonin response. It is localized to the basement membrane and expressed in various structures, including peritoneal cells, ovipositors, gonads, nerve rings, and rectal muscles. It is a homolog of human COL4A1 (type IV collagen alpha 1 chain). To visualize this type of collagen, a green fluorescent protein-collagen fusion strain, emb-9::NLG, was constructed. Intensity of green fluorescence indicates the abundance of this collagen.

[0074] First, OG, GPA, GPO, GPOG, and NAG were dissolved in water to prepare a 1 M solution. This solution was then diluted with OP50 (nutrient-deficient Escherichia coli) to a 100 mM sample solution. emb-9::NLG nematodes were synchronized and grown to the L4 stage (48-60 hours). The nematodes were transferred to FUdR medium (day 0) and cultured at 20°C. The nematodes were fed with sample on days 0, 5, and 13. The nematodes were photographed and counted using an upright fluorescence microscope on days 2, 7, and 15. A minimum of 15 nematodes were counted for each photograph.

[0075] Fluorescence imaging and intensity statistics results are shown in Figure 2. Figure 10 、 11 , 12, wherein, Figure 10 is the expression of EMB-9 in nematodes (day 2 instar), and Figure 10 There are only Mock, GPOG, and NAG groups. Figure 11 is the expression of EMB-9 in nematodes (day 7), and Figure 11 There are only Mock, GPA, and GPOG groups. Figure 12 is the expression of EMB-9 in nematodes (day 15), and Figure 12 There are only Mock, OG, GPO and NAG groups. The Mock group is the blank control group, that is, no active substances OG, GPA, GPO, GPOG, NAG are added.

[0076] Depend on Figure 10 It can be seen that compared with the Mock group, the GPOG group and the NAG group have a significant protective effect on the epidermis of young nematodes.

[0077] Depend on Figure 11 It can be seen that compared with the Mock group, both the GPA group and the GPOG group have a significant protective effect on the epidermis of middle-aged nematodes.

[0078] Depend on Figure 12 It can be seen that compared with the Mock group, the OG group, GPO group and NAG group have a significant protective effect on the epidermis of elderly nematodes.

[0079] Example 6 The expression of COL-12 NLP-29 and EMB-9 by OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG and OG+GPA+GPO+GPOG+NAG were investigated respectively.

[0080] Combinations of OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG, and OG+GPA+GPO+GPOG+NAG were dissolved in water according to the ratios listed in Table 1 to prepare 1 M solutions. These solutions were then diluted with OP50 (nutrient-deficient Escherichia coli) to prepare 100 mM solutions. C. elegans N2 were synchronized and grown to the L4 stage (48-60 hours). The nematodes were then transferred to FUdR medium (to inhibit egg laying) (day 0). Cultures were then incubated at 20°C and fed with food on days 0, 5, and 13. Fluorescence microscopy was used to measure COL-12 and NLP-29 on days 2, 7, and 15. The relevant results are also shown in Table 1. The data for COL-12 (Day 7), COL-12 (Day 15), NLP-29 (Day 2), and NLP-29 (Day 7) are calculated based on the mock group, which served as a blank control group without the active ingredients OG, GPA, GPO, GPOG, or NAG. The corresponding " / " indicates no significant effect compared to the mock group, meaning the results were comparable to or even worse than those in the mock group.

[0081] Table 1 Collagen expression of each composition in Example 6

[0082] As shown in Table 1, OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG and OG+GPA+GPO+GPOG+NAG significantly improved collagen expression and nematode epidermal barrier function under natural aging conditions within a specific proportion range.

[0083] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents, but these modifications or replacements are all within the scope of protection of the present invention.

Claims

1. Use of an active peptide in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: The active peptide has a structural formula of HA-OH, wherein A comprises at least two amino acid residues, and the amino acid residues comprise at least one of Hyp, Gly, Pro, and Ala.

2. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: The product includes at least one of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH.

3. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: Promoting collagen expression includes at least one of the following: (1) promoting the expression of collagen COL-12; (2) promoting the expression of collagen COL-19; (3) promoting the expression of EMB-9 collagen; (4) improving epidermal wrinkles; Improving the skin barrier includes reducing the expression of the antimicrobial protein peptide NLP-29.

4. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: Applicable objects include animals and humans.

5. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: Used to improve the skin barrier of animals or humans during natural aging, and promote collagen expression in humans or animals during natural aging.

6. Use of the active peptide according to claim 3 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: Improving the skin barrier and promoting collagen expression include at least one of the following: (1) H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-12 in middle-aged animals or humans; Or, H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-12 in aged animals or humans; (2) H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-19 in young animals or humans; Or, H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of collagen COL-19 in middle-aged animals or humans; Or, H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-19 in aged animals or humans; (3) H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to reduce the expression of antimicrobial protein peptide NLP-29 in middle-aged animals or humans; (4) H-Hyp-Gly-OH, and / or H-Gly-Pro-Ala-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to improve epidermal wrinkles in animals or humans during middle age; Or, H-Hyp-Gly-OH, and / or H-Gly-Pro-Hyp-OH, and / or H-Gly-Pro-Hyp-Gly-OH are used to improve epidermal wrinkles in animals or humans during old age; (5) H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of basement membrane collagen EMB-9 in young animals or humans; or, H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-Gly-OH are used to increase the expression of basement membrane collagen EMB-9 in middle-aged animals or humans; Alternatively, H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH are used to increase the expression of basement membrane collagen EMB-9 in aged animals or humans.

7. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: The product includes at least two of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH; Or, the product includes at least three of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH; Alternatively, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH.

8. Use of the active peptide according to claim 1 in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: The products include H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH; Or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH; Or, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH; Alternatively, the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH.

9. Use of an active peptide and amino sugar in preparing a product for improving skin barrier and promoting collagen expression, characterized in that: The active peptide has a structural formula of HA-OH, wherein A comprises at least two amino acid residues, and the amino acid residues comprise at least one of Hyp, Gly, Pro, and Ala; The amino sugar includes N-acetyl glucosamine.

10. Use of the active peptide according to any one of claims 1 to 8 for preparing a product that improves the skin barrier and promotes collagen expression, or use of the active peptide and amino sugar according to claim 9 for preparing a product that improves the skin barrier and promotes collagen expression, characterized in that: The product includes at least one of ordinary food, health food, pet food, and medicine.

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