An anti-wrinkle and firming composition and its application

Abstract

This application belongs to the field of daily chemical products and provides an anti-wrinkle and firming composition and its application. The anti-aging and anti-inflammatory composition provided by this application includes a tripeptide, snow algae, and grape flower cell extract. The composition provided by this application combines the tripeptide, snow algae, and grape flower cell extract, with each component working synergistically and promoting each other. It achieves a synergistic effect in promoting the upregulation of Collagen I expression and inhibiting MMP1 expression, thereby exhibiting an anti-wrinkle and firming effect. In addition, the composition provided by this application can also significantly reduce the expression of inflammatory factors, such as IL-8, thus possessing an anti-inflammatory effect.

Description

Technical Field

[0001] This invention relates to the field of daily chemical technology, and in particular to an anti-aging and anti-inflammatory composition, anti-aging and anti-inflammatory products, and their applications. Background Technology

[0002] Skin aging is a gradual physiological process, influenced by various factors such as environment, lifestyle, and disease. With advancements in technology and improved living standards, the demand for functional skincare products is increasing, especially for products with anti-aging properties. Skin aging can be divided into endogenous and exogenous aging. The latter is primarily caused by environmental factors such as reactive oxygen species, free radicals, and ultraviolet radiation. These factors cause cumulative damage to the skin, leading to wrinkles, loss of elasticity, dullness, and other signs of aging.

[0003] Therefore, developing an anti-aging composition is of great practical significance and has broad application prospects for meeting market demands. Summary of the Invention

[0004] In view of this, the technical problem to be solved by the present invention is to provide an anti-wrinkle and firming composition, an anti-wrinkle and firming product and its application. The anti-wrinkle and firming composition provided in this application has a synergistic effect, can significantly promote the upregulation of Collagen I expression, inhibit the expression of MMP1 and IL-8, and has anti-wrinkle, firming and anti-inflammatory effects.

[0005] This application provides an anti-wrinkle and firming composition comprising a tripeptide, snow algae, and grape flower cell extract.

[0006] The anti-wrinkle and firming composition provided in this application includes a tripeptide, particularly palmitoyl tripeptide-5, which promotes collagen production by activating cell signaling pathways, significantly reducing wrinkles, improving skin firmness and radiance, and showing potential in wound healing and tissue repair. This application does not impose any particular limitation on the tripeptide, but preferably on a palmitoyl-modified tripeptide, such as palmitoyl tripeptide-5. Palmitoyl tripeptide-5, with CAS number 623172-55-4, has the amino acid sequence Palm-Lysine-Valine-Lysine-OH, and its terminator can be a hydroxyl group or a salt, such as bis(trifluoroacetate) or mandelate. This application does not impose any particular limitation on the source of the tripeptide, such as palmitoyl tripeptide-5; it can be prepared according to WO2004 / 099237 or directly purchased from the market, such as from Speco, Gakkai, dsm-firmenich Beauty &Care, etc.

[0007] The anti-wrinkle and firming composition provided in this application includes snow algae, which is rich in active ingredients such as carotenoids, antifreeze glycoproteins, and biopolymers, and has anti-aging and antioxidant effects. In some specific implementations, the snow algae includes 1wt%~2wt% Chlamydomonas extract, 80wt%~95wt% isomaltitol, 0.1wt%~1wt% lecithin, and 5wt%~15wt% glycogen. This application does not have any special restrictions on the source of the snow algae; it can be purchased commercially.

[0008] The anti-wrinkle and firming composition provided in this application includes grape flower cell extract, which possesses strong self-renewal and continuous division capabilities, thus exhibiting anti-aging effects. In some specific implementations, the grape flower cell extract is a component obtained from grape flower tissue through induced culture, primarily comprising grape flower cells and also including solvents such as glycerol. In some specific implementations, the grape flower cell extract comprises not less than 20 wt% grape flower cells. This application does not impose any special restrictions on the source of the grape flower cell extract; it can be purchased commercially.

[0009] The composition provided in this application combines tripeptide, snow algae, and grape flower cell extract. The components work synergistically and mutually enhance each other, achieving a synergistic effect in promoting the upregulation of Collagen I expression and inhibiting MMP1 expression, thus exhibiting anti-aging, anti-wrinkle, and firming effects. Furthermore, the composition provided in this application can significantly reduce the expression of inflammatory factors, such as IL-8, demonstrating an anti-inflammatory effect.

[0010] See Figure 1 , Figure 1 This diagram illustrates part of the mechanisms of skin aging. Under ultraviolet radiation, reactive oxygen species (ROS) upregulate the expression of matrix metalloproteinase 1 (MMP1) and inflammatory factors / mediators, leading to collagen degradation and inflammation, thereby causing skin aging.

[0011] In the composition provided in this application, the tripeptide can activate cell signaling pathways, promote collagen production, and inhibit collagen degradation. Grape flower cell extract and snow algae can reduce the damage to skin health caused by free radicals and chronic inflammation, synergistically enhancing the efficacy of the tripeptide and significantly improving the anti-aging effect, achieving a synergistic effect that far surpasses the efficacy of single ingredients in anti-aging. Moreover, the composition provided in this application can penetrate deep into the skin, promoting collagen production, inhibiting collagen degradation, and reducing the expression of inflammation-related genes, thereby comprehensively improving the health of the skin. This comprehensive anti-aging solution overcomes the limitations of single-ingredient effects in existing technologies, providing consumers with a more comprehensive and effective skincare experience. Furthermore, the composition of this application is rationally formulated, achieving significant anti-aging effects at lower dosages, reducing costs, improving cost-effectiveness, and possessing advantages such as high cost-effectiveness and wide applicability.

[0012] In some specific implementations, the mass ratio of the tripeptide, snow algae, and grape flower cell extract is 0.5~15:0.1~1:0.1~5. In some specific implementations, the mass ratio of the tripeptide, snow algae, and grape flower cell extract is 0.6~13:0.3~0.8:0.12~3. In some specific implementations, the mass ratio of the tripeptide, snow algae, and grape flower cell extract is 0.625~12.5:0.5:0.125~2.5.

[0013] This application also provides the application of the anti-aging and anti-inflammatory composition described in the above technical solution in the preparation of anti-inflammatory and / or anti-wrinkle and firming products. The composition provided in this application can be used for anti-aging, anti-wrinkle and firming or anti-inflammation, and can penetrate deep into the skin with significant effects.

[0014] This application also provides the application of the anti-wrinkle and firming composition described above in the preparation of products that promote the expression of type I collagen genes. Specifically, compared with the composition of snow algae single agent, tripeptide and grape flower cell extract, the composition provided in this application produces a synergistic effect in promoting the expression of type I collagen (Collagen I) genes, and has superior anti-aging and anti-wrinkle and firming effects.

[0015] This application also provides the application of the anti-wrinkle and firming composition described in the above-mentioned technical solution in the preparation of products that inhibit MMP1 gene expression. Specifically, compared with the composition of snow algae single agent, tripeptide and grape flower cell extract, the composition provided in this application produces a synergistic effect in inhibiting MMP1 gene expression, and has superior anti-aging and anti-wrinkle and firming effects.

[0016] This application also provides the application of the anti-wrinkle and firming composition described above in the preparation of a product that inhibits IL-8 gene expression. The composition provided in this application can significantly reduce the level of inflammatory factors, such as the expression level of IL-8, thereby exerting an anti-inflammatory effect.

[0017] The present invention also provides an anti-wrinkle and firming product, comprising: the composition described in the above technical solution.

[0018] In some embodiments of the present invention, the above-mentioned products include, but are not limited to, personal care products, pharmaceuticals, etc. Those skilled in the art will understand that, in addition to the compositions described in the above technical solutions, other excipients are also included, such as pharmaceutical excipients or excipients for personal care products.

[0019] In some embodiments of the present invention, the personal care products include: basic care products and / or makeup products.

[0020] In some embodiments of the present invention, the cosmetic products include, but are not limited to:

[0021] (1) Base makeup:

[0022] Foundation / cream: Used to even out skin tone and cover blemishes;

[0023] BB cream / CC cream: A lightweight base makeup product that combines skincare and makeup application;

[0024] Concealer / pen: Used to cover up specific areas such as pimples and dark circles;

[0025] Loose powder / setting powder: Sets makeup and reduces facial shine.

[0026] (2) Eye makeup:

[0027] Eyeshadow: Adds color and dimension to the eyes;

[0028] Eyeliner pencil / liquid / gel: Used to draw lines on the eyes, making them look more vibrant;

[0029] Mascara: Lengthens and thickens eyelashes, enhancing the depth of the eyes;

[0030] Eyebrow pencil / powder / gel: Fill in gaps in eyebrows and create the ideal eyebrow shape.

[0031] (3) Cheek makeup:

[0032] Blush: Adds natural color to the cheeks and improves complexion;

[0033] Contouring powder / stick: Use shading techniques to make facial contours more three-dimensional;

[0034] (4) Lip makeup:

[0035] Lipstick / lip gloss / lip tint: to change or emphasize the color of the lips;

[0036] Lip liner: Defines the clear outline of the lips and prevents lipstick from bleeding.

[0037] (5) Multifunctional cosmetics:

[0038] Highlighter sticks / liquids / powders: Highlight high points on the face (such as the bridge of the nose and cheekbones) to create a luminous effect.

[0039] In addition, there are products designed specifically for special occasions, such as waterproof and sweatproof eyeliners and long-lasting, non-fading lipsticks.

[0040] In some embodiments of the present invention, the basic care products include, but are not limited to:

[0041] Facial cleanser: Gently removes dirt, oil, and makeup residue from the face;

[0042] Cleansing oil / cleansing water / cleansing balm: specially designed to thoroughly remove makeup, especially waterproof cosmetics;

[0043] Toner / lotion: Used after cleansing, it can further cleanse the skin's surface of residue, while replenishing the skin's moisture, restoring the skin's pH balance, and laying a good foundation for the absorption of subsequent skin care products;

[0044] Serum: Contains a high concentration of active ingredients, providing deep nourishment and repair for specific skin problems (such as anti-aging, moisturizing, whitening, etc.);

[0045] Eye cream: Specially designed for the sensitive area around the eyes, it has the effects of reducing fine lines and dark circles and firming the skin around the eyes. The texture is usually light and easily absorbed.

[0046] Day / night lotion or cream: It has functions such as protection, repair and nourishment, and promotes cell regeneration. It can provide the skin with the necessary moisture and lock in the replenished moisture.

[0047] Sunscreen: Used for ultraviolet protection and prevention of photoaging, etc.

[0048] Face masks: provide extra nourishment and care for the skin, such as hydration, pore cleansing, or brightening of the complexion.

[0049] In some specific implementations, the personal care product includes 0.1wt% to 10wt% of the composition described in the above technical solution, preferably 0.5wt% to 9.5wt% of the composition described in the above technical solution, and more preferably 1wt% to 9wt% of the composition described in the above technical solution.

[0050] The composition provided in this application combines tripeptide, snow algae, and grape flower cell extract. These components work synergistically and mutually enhance each other, achieving a synergistic effect in promoting the upregulation of Collagen I expression and inhibiting MMP1 expression, thus exhibiting anti-aging and wrinkle-reducing firming effects. Furthermore, the composition provided in this application can significantly reduce the expression of inflammatory factors, such as IL-8, demonstrating an anti-inflammatory effect. Attached Figure Description

[0051] Figure 1 A diagram illustrating part of the mechanisms of skin aging;

[0052] Figure 2 Summary of Collagen I gene test results;

[0053] Figure 3 Summary of MMP1 gene test results;

[0054] Figure 4 This is a summary of the IL-8 gene test results. Detailed Implementation

[0055] This invention provides an anti-wrinkle and firming composition, an anti-wrinkle and firming product, and their applications. Those skilled in the art can refer to the content herein and appropriately modify the process parameters to achieve the desired results. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0056] The terms “including,” “having,” or “containing,” including the use of their grammatical synonyms, should generally be understood as open-ended and non-restrictive, for example, not excluding other unstated elements or steps, unless otherwise specifically stated or understood from the context.

[0057] It should be understood that the order of the steps or the order in which certain actions are performed is not important as long as the invention remains operational. Furthermore, two or more steps or actions can be performed simultaneously.

[0058] The use of any and all instances or exemplary language such as “e.g.” or “including” in this document is merely intended to better illustrate the invention and is not intended to limit the scope of the invention unless the claims are made. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the invention.

[0059] Furthermore, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Therefore, unless explicitly stated otherwise, it should be understood that all ranges, quantities, values, and percentages used in this disclosure are modified with the word "approximately". Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range.

[0060] This application provides an anti-wrinkle and firming composition comprising a tripeptide, snow algae, and grape flower cell extract.

[0061] The composition provided in this application combines tripeptide, snow algae, and grape flower cell extract. These components work synergistically and promote each other, achieving a synergistic effect in promoting the upregulation of Collagen I expression and inhibiting MMP1 expression, thus exhibiting anti-aging and anti-wrinkle firming effects. Furthermore, the composition provided in this application can significantly reduce the expression of inflammatory factors, such as IL-8, demonstrating an anti-inflammatory effect.

[0062] The present application will be further described below with reference to the embodiments.

[0063] In the following embodiments, snow algae were purchased from Guangzhou Lingyu Trading Co., Ltd.; the tripeptide was palmitoyl tripeptide-5; and the grape flower cell extract was derived from a blended red grape variety from Bordeaux, France, and purchased from Alfa Chemistry.

[0064] Examples 1-3 and Comparative Examples 1-4

[0065] Prepare the composition according to the formulation shown in Table 1:

[0066] Table 1. Formulas provided in the embodiments and comparative examples of this application.

[0067]

[0068] Experimental Example 1

[0069] This test was conducted in the cell laboratory of Shanghai Jiyan Cosmetics Technology Co., Ltd.

[0070] Test method:

[0071] 1. Cell seeding: Select fibroblasts, revive the cells, and seed them into 12-well plates. Incubate overnight in a CO2 incubator (37℃, 5% CO2).

[0072] 2. Test grouping: The experiment set up a blank control group (BC), a negative control group (NC), a positive control group (PC), and a sample group. Each group had 3 replicate wells. The grouping is shown in Table 2. In Table 2, the test samples of each group were the corresponding compositions diluted with DMEM culture medium (Gibco) to make their final concentration meet the dosage requirements of Table 2.

[0073] Table 2 Test Grouping Information

[0074]

[0075] 3. UVA stimulation: According to the test groups, when the cell deposition rate in the 12-well plate reached 40%~50%, except for the blank control group, all other groups were subjected to 14J / cm² stimulation. 2 UVA radiation stimulation.

[0076] 4. Drug administration: After the induction period, drugs were administered to the test groups according to their respective groups. 1 mL of culture medium was added to each well of the blank control group and negative control group, while 1 mL of culture medium containing the corresponding concentration of the test sample was added to each well of the sample group. After drug administration, the 12-well plate was placed in a CO2 incubator (37℃, 5% CO2) and incubated for 24 hours.

[0077] 5. qPCR detection of gene expression (Collagen I): After incubation, the cells were washed, and total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the manufacturer's instructions. Reverse transcription was performed using TOYOBO's ReverTra Ace™ qPCR RTMaster Mix to synthesize cDNA. Using the cDNA as a template, qPCR was performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer (primer sequences are shown in Table 3), 2 μL cDNA template, and 6.4 μL dd H2O. GAPDH was used as an internal reference gene, and the relative expression levels of related genes were calculated using the 2^-ΔΔCt^ method.

[0078] Table 3 Primers for the target gene

[0079]

[0080] 6. Effect value normalization: This test included a positive control group (PC) and a negative control group (NC). The PC group received 100 ng / mL TGF β treatment after stimulation, while the NC group received stimulation without drug administration. The gene upregulation magnitude of PC compared to NC was considered as 1, and the efficacy of other test groups was normalized to the E value.

[0081] The E value can be referenced using the following formula:

[0082] .

[0083] 7. Statistical Analysis of Results: t-tests were used for comparisons between groups, and all statistical analyses were two-tailed. A p-value < 0.05 was considered statistically significant. Results are shown in Table 4 and... Figure 2 Table 4 summarizes the Collagen I gene test results. Figure 2 This is a summary of the Collagen I gene test results.

[0084] Table 4 Summary of Collagen I gene test results

[0085]

[0086] Collagen is an important indicator of skin rejuvenation. (See Table 4 and...) Figure 2 As shown, based on a UVA-induced senescence fibroblast model, the effects of different samples on type I collagen gene expression in fibroblasts were investigated.

[0087] Compared with the BC group, Collagen I expression was significantly downregulated in the NC group, indicating that the stimulation conditions in this test were effective. Compared with the NC group, Collagen I expression was significantly upregulated in the PC group, indicating that the positive control in this test was effective. Compared with the NC group, both Examples 1 and 2 significantly upregulated the gene expression of type I collagen.

[0088] Use the King's formula to verify whether there is synergy:

[0089]

[0090] Compared with Comparative Example 1, Comparative Example 2 and Example 1, Comparative Example 1 is regarded as Agent A, Comparative Example 2 is regarded as Agent B, and Example 1 is Agent A + Agent B, q=39.8>1;

[0091] Compared with Comparative Example 1, Comparative Example 3 and Example 2, Comparative Example 1 is regarded as Agent A, Comparative Example y is regarded as Agent B, and Example 2 is Agent A + Agent B, q=2.8>1.

[0092] Therefore, under the conditions of this test, the combined use of snow algae, tripeptide and grape flower cell extract produced a synergistic effect in promoting the upregulation of Collagen I expression.

[0093] Experimental Example 2

[0094] This test was conducted in the cell laboratory of Shanghai Jiyan Cosmetics Technology Co., Ltd.

[0095] Test method:

[0096] 1. Cell seeding: Select fibroblasts, revive the cells, and seed them into 12-well plates. Incubate overnight in a CO2 incubator (37℃, 5% CO2).

[0097] 2. Test grouping: The experiment set up a blank control group (BC), a negative control group (NC), a positive control group (PC), and a sample group. Each group had 3 replicate wells. The grouping is shown in Table 5. In Table 5, the test samples of each group were the corresponding compositions diluted with DMEM culture medium (Gibco) to make their final concentration meet the dosage requirements of Table 5.

[0098] Table 5 Test Grouping Information

[0099]

[0100] 3. UVA stimulation: According to the test groups, when the cell deposition rate in the 12-well plate reached 40%~50%, except for the blank control group, all other groups were subjected to 14J / cm² stimulation. 2 UVA radiation stimulation.

[0101] 4. Drug administration: After the induction period, drugs were administered to the test groups according to their respective groups. 1 mL of culture medium was added to each well of the blank control group and negative control group, while 1 mL of culture medium containing the corresponding concentration of the test sample was added to each well of the sample group. After drug administration, the 12-well plate was placed in a CO2 incubator (37℃, 5% CO2) and incubated for 24 hours.

[0102] 5. qPCR detection of gene expression (MMP1): After incubation, the cells were washed, and total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the manufacturer's instructions. Reverse transcription was performed using TOYOBO's ReverTra Ace™ qPCR RTMaster Mix to synthesize cDNA. Using the cDNA as a template, qPCR was performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer (primer sequences are shown in Table 6), 2 μL cDNA template, and 6.4 μL dd H2O. GAPDH was used as an internal reference gene, and the relative expression levels of related genes were calculated using the 2^-ΔΔCt^ method.

[0103] Table 6 Primers for the target gene

[0104]

[0105] 6. Effect value normalization: This test included a positive control group (PC) and a negative control group (NC). The PC group received 100 ng / mL TGF β treatment after stimulation, while the NC group received stimulation without drug administration. The gene upregulation magnitude of PC compared to NC was considered as 1, and the efficacy of other test groups was normalized to the E value.

[0106] The E value can be referenced using the following formula:

[0107] .

[0108] 7. Statistical Analysis of Results: t-tests were used for comparisons between groups, and all statistical analyses were two-tailed. A p-value < 0.05 was considered statistically significant. Results are shown in Table 7 and... Figure 3 Table 7 summarizes the MMP1 gene test results. Figure 3 This is a summary of the MMP1 gene test results.

[0109] Table 7 Summary of MMP1 gene test results

[0110]

[0111] Matrix metalloproteinases (MMP1) are a class of enzymes that degrade collagen and are closely related to skin photoaging. In the cosmetics industry, to improve photoaging and protect collagen, active ingredients are usually required to inhibit MMP1 expression. (Table 7 and...) Figure 3The test results for MMP1 are presented, including the mean, standard deviation, p-value analysis, and E-value (power).

[0112] Compared with the BC group, MMP1 expression was significantly upregulated in the NC group, indicating that the stimulation condition was effective. Compared with the NC group, MMP1 expression was significantly downregulated in the PC group, indicating that the positive control was effective. Compared with the NC group, Comparative Examples 1 and 4 could not inhibit MMP1 secretion, while Example 2 significantly reduced MMP1 gene expression. Furthermore, compared with Comparative Examples 1 and 3, Example 2 was significantly better than Comparative Example 3 (p=0.0056), while Algae alone (Comparative Example 1) had no MMP1 inhibitory effect.

[0113] Use the King's formula to verify whether there is synergy:

[0114]

[0115] Compared with Comparative Example 1, Comparative Example 4 and Example 3, Comparative Example 1 is regarded as Agent A, Comparative Example 4 is regarded as Agent B, and Example 3 is Agent A + Agent B, q=23.2>1;

[0116] Compared with Comparative Example 1, Comparative Example 3 and Example 2, Comparative Example 1 is regarded as Agent A, Comparative Example 3 is regarded as Agent B, and Example 2 is Agent A + Agent B, q=1.55>1.

[0117] Therefore, under the conditions of this test, the combined use of snow algae, tripeptide and grape flower cell extract produced a synergistic effect in inhibiting MMP1 expression.

[0118] The above results indicate that the combined use of tripeptide, snow algae, and grape flower cell extract is necessary to produce an MMP1 gene inhibition effect. This synergistic effect is significant for improving MMP1-induced collagen degradation during photoaging and can enhance the product's firming and anti-wrinkle effects.

[0119] Experimental Example 3

[0120] This test was conducted in the cell laboratory of Shanghai Jiyan Cosmetics Technology Co., Ltd.

[0121] Test method:

[0122] 1. Cell seeding: Select fibroblasts, revive the cells, and seed them into 12-well plates. Incubate overnight in a CO2 incubator (37℃, 5% CO2).

[0123] 2. Test grouping: The experiment set up a blank control group (BC), a negative control group (NC), a positive control group (PC), and a sample group. Each group had 3 replicate wells. The grouping is shown in Table 8. In Table 8, the test samples of each group were the corresponding compositions diluted with DMEM culture medium (Gibco) to make their final concentration meet the dosage requirements of Table 8.

[0124] Table 8 Test Grouping Information

[0125]

[0126] 3. UVA stimulation: According to the test groups, when the cell deposition rate in the 12-well plate reached 40%~50%, except for the blank control group, all other groups were subjected to 14J / cm² stimulation. 2 UVA radiation stimulation.

[0127] 4. Drug administration: After the induction period, drugs were administered to the test groups according to their respective groups. 1 mL of culture medium was added to each well of the blank control group and negative control group, while 1 mL of culture medium containing the corresponding concentration of the test sample was added to each well of the sample group. After drug administration, the 12-well plate was placed in a CO2 incubator (37℃, 5% CO2) and incubated for 24 hours.

[0128] 5. qPCR detection of gene expression (IL-8): After incubation, the cells were washed, and total RNA was extracted from each group of cells using the Tiangen Total RNA Extraction Kit, following the manufacturer's instructions. Reverse transcription was performed using TOYOBO's ReverTra Ace™ qPCR RT MasterMix to synthesize cDNA. Using the cDNA as a template, qPCR was performed on a real-time quantitative PCR instrument using SYBR™ Green Realtime PCR Master Mix. The reaction volume was 20 μL, including 10 μL SYBR™ Green Realtime PCR Master Mix, 0.8 μL upstream primer, 0.8 μL downstream primer (primer sequences are shown in Table 9), 2 μL cDNA template, and 6.4 μL dd H2O. GAPDH was used as an internal reference gene, and the relative expression levels of related genes were calculated using the 2^-ΔΔCt^ method.

[0129] Table 9 Primers for the target gene

[0130]

[0131] 6. Effect value normalization: This test included a positive control group (PC) and a negative control group (NC). The PC group received 100 ng / mL TGF β treatment after stimulation, while the NC group received stimulation without drug administration. The gene upregulation magnitude of PC compared to NC was considered as 1, and the efficacy of other test groups was normalized to the E value.

[0132] The E value can be referenced using the following formula:

[0133] .

[0134] 7. Statistical Analysis of Results: t-tests were used for comparisons between groups, and all statistical analyses were two-tailed. A p-value < 0.05 was considered statistically significant. Results are shown in Table 10 and... Figure 4 Table 10 summarizes the IL-8 gene test results. Figure 4 This is a summary of the IL-8 gene test results.

[0135] Table 10 Summary of IL-8 gene test results

[0136]

[0137] Interleukin-8 (IL-8) is a key mediator related to inflammation. Oxidative stress increases IL-8 secretion, leading to the recruitment of inflammatory cells and making it a key parameter of local inflammation. (Table 10 and...) Figure 4 The results of the test on the relative expression level of the IL-8 gene are shown, including the mean, standard deviation, p-value analysis and E value (power).

[0138] Compared with the BC group, IL-8 expression was significantly increased in the NC group, indicating that the stimulation was effective. Compared with the NC group, IL-8 expression was significantly decreased in the PC group, indicating that the positive control was effective. Compared with the NC group, both Comparative Example 1 and Example 2 effectively downregulated IL-8 expression, and the effect of Example 2 was significantly better than that of Comparative Example 1 (p < 0.01), indicating that the combined use of snow algae, tripeptide, and grape flower cell extract can better inhibit the expression of inflammatory genes and has a positive significance for alleviating inflammatory aging.

[0139] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An anti-wrinkle and firming composition comprising a tripeptide, snow algae, and grape flower cell extract, wherein the mass ratio of the tripeptide, snow algae, and grape flower cell extract is 0.625~12.5:0.5:0.125~2.5; The tripeptide is palmitoyl tripeptide-5.

2. The use of the anti-wrinkle and firming composition of claim 1 in the preparation of anti-inflammatory and / or anti-wrinkle and firming products.

3. An anti-wrinkle and firming product, characterized in that, include: The anti-wrinkle and firming composition according to claim 1.

4. The product according to claim 3, characterized in that, For personal care products.

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