Use of a human-derived elastin peptide in the preparation of soothing anti-inflammatory cosmetic products

Abstract

The application proposes an application of a human-derived elastin peptide in preparation of soothing anti-inflammatory cosmetics, and relates to the field of cosmetics. The application screens six elastin peptides, and determines the repair and anti-inflammatory effects thereof, and finally screens the optimal human-derived elastin peptide amino acid sequence, as shown in SEQ ID No: 3. The application applies the human-derived elastin peptide to the fields of skin repair and anti-inflammation. The human-derived elastin peptide can promote skin repair by promoting the proliferation and migration of HaCaT cells, and promoting the expression levels of growth factors and collagen genes of HaCaT cells; meanwhile, the human-derived elastin peptide can inhibit the expression of LPS-induced THP-1 cell inflammatory factors, thereby achieving the dual effects of skin repair and anti-inflammation. The human-derived elastin peptide has a small molecular weight, is beneficial to penetration of the stratum corneum, and improves the delivery efficiency; the human-derived elastin peptide can avoid immunogenicity of animal sources, and can be synthesized by solid-phase peptide synthesis, has a low production cost, and is suitable for industrial production.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, and more specifically, to the application of a human-derived elastin peptide in the preparation of repairing, soothing, and anti-inflammatory cosmetics. Background Technology

[0002] Sensitive skin (SS) specifically refers to a hyperreactive state of the skin under physiological or pathological conditions, manifesting as burning, stinging, itching, and tightness. According to the "Chinese Expert Consensus on the Diagnosis and Treatment of Sensitive Skin," current research suggests that the development of sensitive skin is a complex process involving the skin barrier, neurovascular system, and immune inflammation. Under the interaction of internal and external factors, impaired skin barrier function leads to increased sensory nerve afferent signals, resulting in enhanced skin reactivity to external stimuli, which in turn triggers an immune inflammatory response. Therefore, the vicious cycle between epidermal barrier dysfunction and excessive inflammatory response is a critical issue for sensitive skin and related skin aging problems.

[0003] The Chinese Journal of Dermatology, in its "Guidelines for the Application of Soothing and Moisturizing Skincare Products in Sensitive Skin," points out that simply repairing or reducing inflammation alone cannot fundamentally cure sensitive skin. Therefore, the development of skincare and cosmetic products for sensitive skin should prioritize active ingredients that possess both barrier repair and anti-inflammatory soothing properties to achieve long-term stability for sensitive skin.

[0004] Therefore, most cosmetics currently on the market formulated for sensitive skin combine ingredients with barrier repair and anti-inflammatory soothing effects, such as ceramides, squalane, panthenol, asiaticoside, purslane extract, and elastin peptides. Among these, elastin peptides are rapidly expanding in application due to their low allergenicity and suitability for sensitive skin pain points. They can repair sensitive skin by promoting angiogenesis, collagen and elastin synthesis, keratinocyte proliferation and migration, tissue remodeling, and reducing skin inflammation, ultimately restoring the integrity of the skin's structure and function.

[0005] However, existing elastin peptides have relatively limited efficacy, mostly possessing only single repair or anti-inflammatory effects. For example, Chinese patent CN120241520A discloses a composition containing elastin peptides, which only mentions promoting skin wound healing without mentioning anti-inflammatory effects. CN112869150A discloses a method that combines astaxanthin, bonito elastin peptides, fish collagen peptides, natural vitamin C, and hyaluronic acid to prevent skin inflammation caused by medium- and long-wave ultraviolet (UVB-UVA) radiation. Furthermore, research by Qiaoli Wu et al. demonstrated that LGVGSPPGK, GFDPSR, and PGAPGGGFDI in enzymatically extracted bonito elastin peptides (STEP) alleviate IL-1β-induced osteoarthritis and promote chondrocyte regeneration by regulating inflammatory factors and the JAK2 / STAT3 signaling pathway; however, no data have proven that STEP elastin peptides have a repair effect on skin barrier cells.

[0006] Furthermore, most existing elastin peptides are synthesized using animal-derived hydrolysis extraction methods, which involves a complex overall preparation process and high production costs. Moreover, applying animal-derived elastin peptides directly to the skin as cosmetics carries the risk of inducing rashes, itching, and urticaria.

[0007] Based on the above research background, this invention focuses on the repair and anti-inflammatory activities of elastin peptides. By screening the characteristic sequences of six mammalian tropoelastin, a single human elastin peptide with both skin repair and anti-inflammatory activities was finally obtained. Summary of the Invention

[0008] The purpose of this invention is to screen six elastin peptides for their effects on the proliferation and migration of HaCaT cells, and then evaluate whether the elastin peptide with the best effect promotes the expression levels of growth factor genes and collagen genes in HaCaT cells. In addition, a THP-1 cell inflammation model is established to evaluate the anti-inflammatory effect of the elastin peptide, and to provide the application of this elastin peptide in the preparation of repair, soothing and anti-inflammatory cosmetics.

[0009] The human elastin peptide obtained in the final screening of this invention promotes the proliferation and migration of HaCaT cells, and promotes the expression levels of growth factor genes and collagen genes in HaCaT cells; it also inhibits the expression levels of inflammatory factors in LPS-induced THP-1 cells, thereby achieving a dual effect of skin repair, soothing and anti-inflammation.

[0010] After screening six elastin peptides, this invention provides a human elastin peptide with better performance, which is exon 26 (Domain-26) of human tropoelastin, and its amino acid sequence is shown in SEQ ID No:3.

[0011] To achieve the above objectives, the present invention employs the following technical solutions.

[0012] On the one hand, this application provides an application of human elastin peptide in the preparation of repairing, soothing and anti-inflammatory cosmetics, wherein the amino acid sequence of the human elastin peptide is shown in SEQ ID No:3.

[0013] In the applications described above, the concentration of human elastin peptides is 0-50 μg / mL.

[0014] As described above, in this application, human elastin peptides promote skin repair by stimulating the expression of growth factor and collagen genes in HaCaT cells.

[0015] As described above, in this application, human elastin peptides enhance the skin's anti-inflammatory capacity by inhibiting the expression levels of inflammatory factors in LPS-induced THP-1 cells.

[0016] On the other hand, embodiments of the present invention also provide a repairing, soothing, and anti-inflammatory cosmetic as described above. The components of the cosmetic, by mass fraction, include: EDTA-2 Na 0.01%-0.08%, glycerin 2%-6%, methyl ester 0.1%-0.3%, phenoxyethanol 0.1%-0.3%, emulsifier 1%-3%, thickener 2%-5%, triglyceride (ethylhexanoate) 2%-5%, ethylhexyl palmitate 5%-10%, propyl ester 0.1%-0.5%, moisturizer 1%-5%, human elastin peptide 0.1%-1%, and the balance being deionized water.

[0017] Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects: 1. This invention is the first to apply human elastin peptide (AGVPGLGVG) to the field of skin repair and anti-inflammation. It promotes skin repair by promoting the proliferation and migration of HaCaT cells and promoting the expression levels of growth factor genes and collagen genes in HaCaT cells. At the same time, it can also improve the anti-inflammatory effect of the skin by inhibiting the expression level of inflammatory factors in LPS-induced THP-1 cells. 2. The human elastin peptide (AGVPGLGVG) provided by this invention has a small molecular weight of only 725.84 Da, which is beneficial for penetrating the stratum corneum and improving delivery efficiency; it can also be synthesized through solid-phase peptide (SPPS), which has a low production cost and is suitable for industrial production. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a schematic diagram illustrating the effect of different concentrations of AGVPGLGVG peptide on the migration of HaCaT cells in Example 1 of the present invention. Figure 2 This is a graph showing the expression level of COL17A under the influence of different concentrations of AGVPGLGVG peptide in Example 1 of the present invention. Figure 3 This is a graph showing the expression levels of VEGF under the influence of different concentrations of AGVPGLGVG peptide in Example 1 of the present invention. Figure 4 The expression levels of inflammatory factors in LPS-induced THP-1 cells under the influence of different concentrations of AGVPGLGVG peptide in Example 2 of this invention; Figure 5 This is a comparative diagram of the skin damage repair experiments of mice in different groups in Example 3 of the present invention; Figure 6 The results of TEWL measurement in each group of mice in Example 3 of this invention; Figure 7 The results of H&E staining of mouse skin tissue samples in each group in Example 3 of this invention are shown. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0021] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present invention will now be described in detail with reference to specific embodiments.

[0022] In the following examples, the English characters represent the following meanings: EDTA-2 Na: Disodium ethylenediaminetetraacetate; HaCaT: Immortalized human keratinocytes; VEGF: Vascular endothelial growth factor; COL17A: Type XVII collagen; THP-1: A human acute monocytic leukemia cell line; PMA: Phloride; IL-6: Interleukin-6; IL-1β: Interleukin-1β; COX-2: Cyclooxygenase-2; TNF-α: Tumor necrosis factor-α.

[0023] Example 1 This embodiment provides six types of elastin peptides, wherein the amino acid sequences of the elastin peptides are as follows: SEQ ID No:1 is VGVAPG. Specifically: Valine-Gly-Val-Ala-Pro-Gly. SEQ ID No:2 is VPGVG. Specifically: Valine-Proline-Gly-Val-Gly.

[0024] SEQ ID No:3 is AGVPGLGVG. Specifically, it is: Alanine-glycine-valine-proline-glycine-leucine-glycine-valine-glycine (Ala-Gly-Val-Pro-Gly-Leu-Gly-Val-Gly).

[0025] SEQ ID No:4 is PGAIPG. Specifically: Proline-Glycine-Alanine-Ile-Pro-Gly.

[0026] SEQ ID No:5 is PGVGVA. Specifically, it is: Proline-Glycine-Valine-Glycine-Valine-Ala.

[0027] SEQ ID No:6 is AGVPGFGVG. Specifically: alanine-glycine-valine-proline-glycine-phenylalanine-glycine-valine-glycine (Ala-Gly-Val-Pro-Gly-Phe-Gly-Val-Gly).

[0028] In addition, this embodiment also verified the effects of six elastin peptides on HaCaT cells, including the following experimental protocol: (I) Effects of six elastin peptides on HaCaT cell proliferation: 1) Remove the frozen HaCaT cell cryovials from the liquid nitrogen container and quickly place them in a 37°C constant temperature water bath. Shake rapidly until completely thawed (about 1-2 min), avoiding repeated freeze-thaw cycles. Centrifuge at 800 rpm for 5 min and discard the supernatant. Resuspend the cell pellet in 2 mL of complete culture medium, transfer it to a T25 culture flask, add complete culture medium to 5 mL, shake gently, and place in a 37°C, 5% CO2 incubator for static culture. Observe the cell adhesion after 24 h, replace with fresh complete culture medium, and remove any unattached dead cells.

[0029] 2) When HaCaT cells grow to 80%-90% (density) of the bottom area of ​​the culture flask, passage them. Discard the old culture medium in the culture flask, gently wash the cell surface twice with PBS buffer, and aspirate the PBS. Add 1 mL of 0.25% trypsin-EDTA digestion solution, place in a 37℃ incubator for 8 min, and observe under an inverted microscope. When the cells become round and begin to detach, immediately add 1 mL of complete culture medium to stop the digestion. Gently pipette the cells to disperse them evenly into a single-cell suspension. Transfer the cell suspension to a sterile centrifuge tube, centrifuge at 800 rpm for 5 min, and discard the supernatant. Resuspend the cells in complete culture medium, passage them in flasks at a ratio of 1:2 to 1:4, label the cell names, passage number, and culture date, and continue culturing in an incubator. Replace with fresh complete culture medium every 3-4 days, passage at least 3 times, and select cells with good growth and uniform morphology for subsequent detection experiments.

[0030] 3) HaCaT cells in the logarithmic growth phase were digested with 0.25% trypsin-EDTA digestion solution, and then digestion was terminated by adding complete culture medium. The cells were pipetted to prepare a single-cell suspension. Cells were counted using a cell counting chamber to ensure cell viability ≥95%. Then, 5000 cells / well were seeded into 96-well cell culture plates. Only 100 μL of complete culture medium was added to the outermost well of the 96-well plate (without cell seeding) as blank control wells. The seeded 96-well plates were placed in a 37℃, 5% CO2 incubator for 24 h to allow the cells to adhere completely and return to normal growth.

[0031] 4) After cell adhesion, cells were starved in serum-free medium for 4 h. Different concentrations of samples were added and diluted with 2% serum medium (concentrations of 3.125 μg / mL, 6.25 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, and 100 μg / mL). The blank control group was added with the same volume of medium containing 2% serum, and the positive control group was added with medium containing 10% serum. All experimental groups, blank control group, and positive control group were incubated at 37℃ in a 5% CO2 incubator for 24 h. After 24 h and 48 h of treatment, 10 μL of CCK8 was added to each well, and cell viability was measured at 450 nm and 630 nm wavelengths after 2 h. Cell viability was calculated (three biological replicates were set up for each experimental group). The experimental results are shown in Table 1, where * indicates P<0.05 compared with the control group; ** indicates P<0.01 compared with the control group; *** indicates P<0.001 compared with the control group; and **** indicates P<0.0001 compared with the control group.

[0032] Table 1. Effects of six elastin peptides on HaCaT cell proliferation (24 h)

[0033] As shown in Table 1, at a concentration of 50 μg / mL, all six elastin peptides significantly promoted the proliferation of HaCaT cells after 24 h of treatment. However, AGVPGLGVG peptide significantly promoted the proliferation of HaCaT cells starting at a concentration of 6.25 μg / mL after 24 h of treatment, demonstrating that AGVPGLGVG peptide had the best promoting effect on HaCaT cell proliferation.

[0034] (II) Effects of six elastin peptides on HaCaT cell migration: HaCaT cells cultured according to protocol (I) were used at a rate of 3 × 10⁻⁶. 5Cells were seeded per well in 12-well plates. After cell adhesion, the wells were scratched with a pipette tip, and different concentrations of six elastin peptides (50 μg / mL) were added for 24 h and 48 h. 10% FBS was used as a positive control. Furthermore, in cell migration assays, if cell proliferation is not inhibited, the observed increase in the number of migrating cells may be influenced by an increased total cell number due to accelerated cell proliferation. Therefore, to distinguish between "migration" and "proliferation," the cell proliferation inhibitor CDK4 / 6 (palbociclib) 1 μM was added to ensure that the experimental results reflect the actual change in migration ability, rather than differences in cell number. Photos were taken and the scratch healing rate was calculated every 24 hours. The experimental results are shown in Table 2. All six elastin peptides at a concentration of 50 μg / mL significantly promoted HaCaT cell migration after 48 h of treatment; however, after 24 h of treatment, only AGVPGLGVG peptide at a concentration of 50 μg / mL significantly promoted HaCaT cell migration. Therefore, AGVPGLGVG peptide showed the best effect in promoting HaCaT cell migration. P<0.05; ** indicates P<0.01 compared with the control group; *** indicates P<0.001 compared with the control group; **** indicates P<0.0001 compared with the control group.

[0035] Table 2. Effects of six elastin peptides on the migration of HaCaT cells (24 h, 48 h).

[0036] Therefore, the next step was to further screen the specific migration-promoting concentration of AGVPGLGVG peptide. Different concentrations of AGVPGLGVG peptide (6.25 μg / mL, 12.5 μg / mL, and 25 μg / mL) were added for treatment for 24 h and 48 h. VEGF was used as a positive control. Photos were taken and recorded every 24 hours, and the scratch healing rate was calculated. The experimental results are as follows: Figure 1 As shown in Table 3, at a concentration of 25 μg / mL, AGVPGLGVG peptide significantly promoted HaCaT cell migration after 48 h of treatment, demonstrating that AGVPGLGVG peptide at 25 μg / mL can promote HaCaT cell migration. * indicates P < 0.05 compared to the control group; ** indicates P < 0.01 compared to the control group; *** indicates P < 0.001 compared to the control group; **** indicates P < 0.0001 compared to the control group.

[0037] Table 3. Effects of AGVPGLGVG peptide on HaCaT cell migration (24 h, 48 h)

[0038] (III) Effects of AGVPGLGVG peptide on the expression levels of growth factor and collagen genes in HaCaT cells Having determined that the AGVPGLGVG peptide exhibits the best proliferation and migration effects, the next step is to evaluate its impact on the expression levels of growth factors and collagen genes in HaCaT cells. HaCaT cells cultured according to protocol (I) were used at a rate of 5 × 10⁶ cells / year. 5 Cells were seeded per well in 6-well plates. After cell adhesion, AGVPGLGVG peptides were added at concentrations of 6.25 μg / mL, 12.5 μg / mL, and 25 μg / mL, respectively. RNA was extracted using the Trizol method, and then cDNA was synthesized by reverse transcription. The expression levels of growth factor and collagen genes in HaCaT cells were detected by qPCR. VEGF was used as a positive control. The qPCR reaction system and primer sequences used are shown in SEQ ID No. 7-SEQ ID No. 12. The experimental results are as follows: Figure 2 , Figure 3 As shown in Table 4, it can be seen that AGVPGLGVG peptide can significantly promote the expression levels of growth factor and collagen genes in HaCaT cells at concentrations of 6.25 μg / mL and 12.5 μg / mL, respectively.

[0039] Table 4: qPCR primer sequences

[0040] Then, qPCR detection was performed based on the primer sequences described above, as follows (three replicates were set up for each sample):

[0041] Table 4. Effects of AGVPGLGVG peptide on the expression levels of growth factor genes in HaCaT cells (24 h).

[0042] Where * indicates P < 0.05 compared to the control group; ** indicates P < 0.01 compared to the control group; *** indicates P < 0.001 compared to the control group; **** indicates P < 0.0001 compared to the control group.

[0043] In summary, among the six elastin peptides, AGVPGLGVG peptide showed the best effect in promoting HaCaT cell proliferation and migration, and also increased the expression levels of COL17A and VEGF in HaCaT cells. HaCaT cells are crucial cells for rebuilding the epidermal barrier, further demonstrating that AGVPGLGVG peptide, as a cosmetic ingredient, has a repairing effect on damaged skin, and this repair effect is superior to the other five screened elastin peptides.

[0044] Example 2 This embodiment investigates the effect of AGVPGLGVG peptide on THP-1 cells in Example 1, further exploring whether AGVPGLGVG peptide possesses anti-inflammatory effects, including the following methods: (a) Arrange THP-1 cells at 8 x 10 5 Cells were seeded per well in 6-well plates. After LPS induction for 48 h, cells were treated with AGVPGLGVG peptide at concentrations of 5 μM, 10 μM, 20 μM, and 1 μg / mL LPS for 4 h, respectively. 10 μM dexamethasone (DXM) was used as a positive control. Cells were collected, RNA was extracted, reverse transcribed, and then the expression levels of inflammatory factors were detected by qPCR. Experimental results are as follows: Figure 4 As shown in Table 5, compared with the Control group, the mRNA expression levels of IL-6, TNF-α, IL-1β, and COX-2 in the Model group were significantly increased, indicating that the inflammation model was successfully constructed. Compared with the model group, the AGVPGLGVG peptide concentration of only 5 μM significantly inhibited the gene expression levels of IL-6 and COX-2; at 10 μM, it significantly inhibited the gene expression level of IL-1β; and at 20 μM, it significantly inhibited the gene expression level of TNF-α. This demonstrates that the AGVPGLGVG peptide can significantly inhibit the expression levels of inflammatory factors IL-6, TNF-α, COX-2, and IL-1β within the concentration range of 5-20 μM. Wherein, ####: P < 0.0001 compared with the control group; *: P < 0.05 compared with the model group; **: P < 0.01 compared with the model group; ***: P < 0.001 compared with the model group; ****: P < 0.0001 compared with the model group.

[0045] Table 5. Effects of AGVPGLGVG peptide on the expression levels of inflammatory cytokine genes in LPS-induced THP-1 cells.

[0046] In conclusion, AGVPGLGVG peptide can significantly inhibit the expression level of inflammatory factors in LPS-induced THP-1 cells, further demonstrating that AGVPGLGVG peptide can be used as a cosmetic ingredient with anti-inflammatory effects.

[0047] Example 3 This embodiment provides a repairing, soothing, and anti-inflammatory cosmetic, the composition of which includes: EDTA-2 Na 0.05%, glycerin 4%, methyl ester 0.2%, phenoxyethanol 0.2%, emulsifier 2%, thickener 4%, glycerin tri(ethylhexanoate) 5%, ethylhexyl palmitate 7%, propyl ester 0.1%, moisturizer 2%, human elastin peptide 0.1%, and the balance being deionized water.

[0048] In addition, the emulsifier chosen is BS-1768, whose core components are cetearyl alcohol and cetearyl glucoside, which are nonionic natural glycosyl liquid crystal emulsifiers. The thickener chosen is cetearyl alcohol, and the humectant is ethyl glucoside or a polyol humectant.

[0049] Furthermore, this embodiment also provides a method for preparing the cosmetic, including the following steps: S1: Add an appropriate amount of deionized water (500 mL as an example) to the emulsifying pot and stir at a low speed of 30-50 rpm; then add 0.5 mL of EDTA-2 Na and 40 mL of glycerol in sequence and stir until completely dissolved; then add 2 mL of methyl ester and 2 mL of phenoxyethanol, continue stirring until uniform, turn on the heating, raise the temperature to 75±1℃, keep it at this temperature for 10 min to ensure that the water-soluble raw materials are completely dissolved and there are no particulate residues, and obtain phase A (aqueous phase) solution; S2: In a separate oil phase dissolving vessel, add 40 mL of cetearyl alcohol, 50 mL of triethylhexanoate, and 70 mL of ethylhexyl palmitate. Stir at a low speed of 30-50 rpm and heat to 78±2℃. After complete melting, add 20 mL of BS-1768 and stir until completely dissolved. Then add 1 mL of propyl ester, stir to mix well, and keep warm to 75±1℃ to obtain phase B (oil phase). S3: Continue stirring the emulsifier at a low speed of 30-50 rpm, slowly pumping the oil phase into the water phase while stirring throughout to prevent the oil phase from floating. After the two phases are mixed, transfer them to a homogenizer and gradually increase the speed to 300-350 rpm, homogenizing for 10-15 minutes. During homogenization, observe the material in real time: it is qualified if it is fine and milky white, without particles or stratification. If particles appear, extend the homogenization time by 3-5 minutes. After homogenization, turn off the homogenizer and allow it to cool naturally. When the material temperature drops to 35±2℃, add 20mL of humectant and stir until completely dispersed. Continue cooling. When the temperature drops to 25±2℃, add 10μg of human elastin peptide and add deionized water to make the volume reach 1 L, stirring until completely dissolved. Then let it stand to remove bubbles to obtain the cosmetic product, named elastin peptide face cream.

[0050] Comparative Example The preparation methods for this comparative example and Example 3 are basically the same, the only difference being that it does not contain human-derived elastin peptides. The resulting cosmetic product is named elastin peptide control cream.

[0051] Animal experiments were conducted using the elastin peptide face cream from Example 3 and a comparative elastin peptide control face cream to test the repair effect of the cosmetics on the damaged skin barrier in mice. The procedures included the following: First, a mouse animal model was established, including: 6-week-old SPF-grade BALB / c male mice, which were acclimatized for 7 days and then randomly divided into 4 groups (blank group, model group, elastin peptide control cream group, and elastin peptide cream group, 6 mice in each group). One day before modeling, all mice underwent hair removal on a localized area of ​​their backs. The next day, except for the blank group, all other groups of mice were repeatedly adhered to the hair-removed area of ​​their backs with strong adhesive tape 8-10 times to construct a mouse back epidermal barrier damage model. Appropriate concentrations of sample were applied to the tape-attached areas of each treatment group, while the blank and model groups were given appropriate amounts of physiological saline. The application was repeated twice a day, and photographs were taken for recording. After 7 days of continuous treatment, transdermal water loss (TEWL) was measured at the tape-attached areas of the mice's backs using a Vapo Meter transdermal water meter. The mice were then sacrificed, and skin tissue from the lesion area on the back was collected and prepared into pathological sections for H&E staining analysis to observe the histopathological manifestations of the damaged epidermal barrier.

[0052] The growth of the epidermis in each group of mice is as follows: Figure 5 As shown, on day 7, the skin in the blank group remained smooth and intact, without any damage or scabs, representing the baseline state of healthy skin. The model group showed large areas of dark brown scabs and damage, with the worst skin condition, indicating successful modeling and that without intervention, skin damage would continue to worsen and would be difficult to repair on its own. The elastin peptide control cream group also showed obvious scabs and damage, similar to the damage level in the model group, indicating that the blank cream alone did not have a significant repair effect and the damage could not be effectively improved. On day 7, the skin condition of the elastin peptide cream group was significantly better than that of the model group and the control cream group, with smaller scabs, lighter color, and better skin integrity, indicating that the cream containing elastin peptides can significantly promote the repair of skin damage and reduce inflammation and scabs.

[0053] One objective indicator for assessing sensitive skin is transepidermal water loss (TEWL), which indirectly reflects the function of the skin's stratum corneum barrier. This value is often elevated in sensitive skin. One function of the epidermal barrier is to prevent excessive water loss from the skin. When the integrity of the epidermis is compromised, the epidermal barrier function is also damaged, leading to increased transepidermal water loss. It can directly reflect the degree of damage to the epidermal barrier function and indirectly reflect the repair status after damage. The TEWL test results of mice in each group in this protocol are as follows: Figure 6As shown, the blank group had the lowest TEWL value, representing the normal level of healthy skin. The model group had a significantly higher TEWL value, indicating successful skin barrier modeling, severe damage, a significant decrease in water retention capacity, and increased moisture loss. The TEWL value of the elastin peptide control cream group was slightly lower than that of the model group, indicating that using only the blank cream base had a weak moisturizing effect, but the degree of improvement was limited. The TEWL value of the elastin peptide cream group was significantly lower, indicating that the cream containing elastin peptides could significantly reduce TEWL, that is, significantly repair the damaged skin barrier. This repair effect was far superior to that of the blank cream, proving that the effect came from the active ingredient of elastin peptides, rather than the cream base itself.

[0054] In addition, since damage to the epidermal barrier leads to varying degrees of pathological changes in skin tissue structure, the H&E staining results of the skin tissues of mice in each group in this protocol are as follows: Figure 7 As shown, compared to the control group, the epidermal structure of mice was damaged and the epidermis was significantly thickened after repeated application of tape to the back of the mice, exhibiting hyperplastic symptoms. However, after continuous application of elastin peptide cream, the hyperplastic symptoms of the mice's epidermis were significantly improved, and the thickened epidermis gradually thinned and returned to normal. This result shows that elastin peptide cream can alleviate hyperplastic symptoms of the epidermis in mice and promote the repair of epidermal barrier damage.

[0055] In summary, this invention provides an application of human elastin peptides in the preparation of repairing, soothing, and anti-inflammatory cosmetics. It is the first time that human elastin peptides (AGVPGLGVG) have been applied to both skin repair and anti-inflammation. It promotes skin repair by increasing the proliferation and migration of HaCaT cells and by increasing the expression levels of growth factor genes and collagen genes in HaCaT cells. Simultaneously, it enhances the anti-inflammatory effect by inhibiting the expression of inflammatory factors in LPS-induced THP-1 cells. Furthermore, animal experiments have demonstrated that the prepared elastin peptide cream can alleviate epidermal hyperplasia-like symptoms in mice and promote the repair of epidermal barrier damage.

[0056] The human elastin peptide (AGVPGLGVG) provided by this invention has a small molecular weight, which is conducive to penetrating the stratum corneum and improving delivery efficiency; it can also be synthesized by solid-phase peptide (SPPS), which has a low production cost and is suitable for industrial production.

[0057] The embodiments described above are some, but not all, embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. The application of a human-derived elastin peptide in the preparation of repairing, soothing, and anti-inflammatory cosmetics, characterized in that, The amino acid sequence of the human elastin peptide is shown in SEQ ID No:

3.

2. The application according to claim 1, characterized in that, The application concentration of the human elastin peptide is 0-50 μg / mL.

3. The application according to claim 1 or 2, characterized in that, The human-derived elastin peptide promotes skin repair by stimulating the expression of growth factor and collagen genes in HaCaT cells.

4. The application according to claim 1 or 2, characterized in that, The human elastin peptide enhances the skin's anti-inflammatory capacity by inhibiting the expression levels of inflammatory factors in LPS-induced THP-1 cells.

5. A repairing, soothing, and anti-inflammatory cosmetic product as described in claim 1, characterized in that, The cosmetic composition, by mass fraction, includes: EDTA-2Na 0.01%-0.08%, glycerin 2%-6%, methyl ester 0.1%-0.3%, phenoxyethanol 0.1%-0.3%, emulsifier 1%-3%, thickener 2%-5%, triglyceride (ethylhexanoate) 2%-5%, ethylhexyl palmitate 5%-10%, propyl ester 0.1%-0.5%, moisturizer 1%-5%, human elastin peptide 0.1%-1%, and the balance being deionized water.

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