Preparation method and application of calcium-containing fermentation lysate with anti-aging efficacy

Abstract

This invention belongs to the field of cosmetic technology, specifically relating to a method for preparing a calcium-containing fermented lysate with anti-aging effects and its application. This invention is the first to prepare a fermented lysate with high calcium content. Experiments have shown that this calcium-containing fermented lysate can significantly promote the relative gene expression levels of collagen and elastin, exhibiting significant anti-wrinkle effects. Simultaneously, this calcium-containing fermented lysate can effectively inhibit the activity of elastase, resulting in a significant firming effect. Furthermore, this invention applies the calcium-containing fermented lysate to cosmetics. The interaction of the components in this calcium-containing fermented lysate composition not only improves the skin's absorption rate but also effectively enhances the stability of the composition, thus facilitating the stable exertion of its anti-aging effects. This makes it a relatively ideal anti-aging cosmetic.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a method for preparing and applying a calcium-containing fermented lysate with anti-aging effects. Background Technology

[0002] Aging is an irreversible decline in physiological structure and function that occurs with age, specifically divided into endogenous and exogenous aging. Endogenous aging refers to natural aging that occurs with increasing age, while exogenous aging mainly refers to photoaging caused by sun exposure. The various fibrous network structures in the skin act like a stable steel reinforcement structure, supporting the entire skin tissue and giving it tensile strength and elasticity. However, as the skin ages, these "steel bars" gradually break and collapse, causing the skin to lose elasticity and develop wrinkles.

[0003] Currently, various anti-aging cosmetics have emerged on the market to combat skin aging, but many problems remain. For example, anti-aging cosmetics often have complex ingredients but low concentrations, failing to achieve the desired anti-aging effect; their ingredients are often too simple or too concentrated, easily causing skin irritation; and the active ingredients in anti-aging cosmetics are mostly unstable, easily deactivated, and difficult to absorb through the skin, resulting in low bioavailability. In recent years, with consumers increasingly focusing on "green, natural, and safe" cosmetics, the development of fermented cosmetics has made significant progress. Among these, microbial fermentation products are becoming a research hotspot as cosmetic raw materials, with their core advantages lying in enhancing the efficacy of active ingredients, improving safety, and promoting green manufacturing.

[0004] Patent document CN108478459A discloses an eye serum containing Lactobacillus fermentation lysate and its preparation method. This eye serum mainly consists of Lactobacillus fermentation lysate, tea extract, pomegranate peel extract, caffeine, royal jelly peptides, oat β-glucan, buckwheat seed extract, ceramides, emulsifiers, and solvents. The eye serum uses tea extract, pomegranate peel extract, and caffeine as the main framework for its anti-aging and antioxidant effects. Combined with Lactobacillus fermentation lysate to enhance skin cell energy and improve skin absorption, and with royal jelly peptides, oat β-glucan, buckwheat seed extract, and ceramides as the nutritional framework, it promptly replenishes nutrients and moisture to the eye area, enhancing the eye serum's anti-aging, wrinkle-reducing, antioxidant, and nutrient-replenishing effects.

[0005] Patent document CN114891846A discloses a Bifida ferment lysate and its application in whitening and anti-aging cosmetics. This Bifida ferment lysate contains Bifida polysaccharide, which has terminal glucose and galactose residues. The molecular structure of this polysaccharide mainly comprises glucose and galactose units, and also includes glucosamine and galactosamine. This Bifida polysaccharide not only has significant antioxidant and tyrosinase-inhibiting properties, but also promotes fibroblast growth and proliferation, thus exhibiting whitening and anti-aging effects.

[0006] However, the mechanisms currently used in anti-aging cosmetics mainly involve resisting free radicals or antioxidant effects, which are unstable and cannot achieve long-lasting anti-aging results. Therefore, researching and developing cosmetics with high stability and significant anti-aging efficacy remains a challenge. Summary of the Invention

[0007] To address the shortcomings of existing technologies, this invention provides a method for preparing and applying a calcium-containing fermented lysate with anti-aging effects. This invention obtains a calcium-containing fermented lysate through bio-fermentation, and then applies this lysate to a composition containing polyglycerol-10 laurate, PEG-40 hydrogenated castor oil, lecithin, and palmitoyl pentapeptide-4. The intracellular calcium in this composition can promote epidermal cell proliferation through the calcium-regulating pathway and, synergistically, the palmitoyl pentapeptide-4 polypeptide to construct the epidermal and dermal reticular structure, supporting the skin and achieving an anti-aging effect. Simultaneously, the synergistic effect of the polyglycerol-10 laurate, PEG-40 hydrogenated castor oil, and lecithin effectively promotes skin absorption and improves the stability of the composition, resulting in a significant anti-aging effect.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] This invention provides a method for preparing a calcium-containing fermented lysate with anti-aging effects, comprising the following steps:

[0010] Step S1: Dissolve each culture medium component in pure water, sterilize, and then place it in a clean bench to inoculate the bacterial culture. Then ferment it in a constant temperature shaking shaker at a temperature of 25~30 ℃ and a speed of 120~160 rpm for 18~24 h to obtain the fermentation broth.

[0011] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 10,000 to 12,000 rpm for 25 to 35 minutes. Take the precipitate and perform low-temperature microfluidic extraction and crushing to obtain calcium-containing fermentation lysate.

[0012] Furthermore, the culture medium in step S1 consists of the following components:

[0013] The composition includes: glucose 90-150 g / L, sodium dihydrogen phosphate 0.4-0.8 g / L, disodium hydrogen phosphate 0.1-0.6 g / L, magnesium sulfate 0.05-0.4 g / L, peptone 1-16 g / L, and water 800-950 g / L. The peptone is bovine bone peptone, which contains calcium for use by sake yeast.

[0014] Preferably, the culture medium in step S1 consists of the following components:

[0015] The mixture contained 110 g / L glucose, 0.4 g / L sodium dihydrogen phosphate, 0.2 g / L disodium hydrogen phosphate, 0.15 g / L magnesium sulfate, 8 g / L peptone, and 881.25 g / L purified water. The peptone was bovine bone peptone.

[0016] Furthermore, the strain in step S1 is sake yeast, and the inoculation amount of sake yeast is 5-10%.

[0017] Furthermore, the conditions for low-temperature microjets to extract and break down the material in step S2 are: extraction for 30 to 50 minutes at a temperature of 20 to 30 ℃ and a flow rate of 15 to 20 L / min.

[0018] Furthermore, the present invention also claims protection for the calcium-containing fermented lysate obtained by the method for preparing the calcium-containing fermented lysate with anti-aging effects.

[0019] In addition, the present invention also provides the application of the calcium-containing fermented lysate with anti-aging effects in the preparation of anti-aging cosmetics.

[0020] Furthermore, the cosmetics include gels, facial cleansers, toners, serums, creams, lotions, and moisturizers.

[0021] Furthermore, the present invention also provides a composition with anti-aging effects, comprising the following components and their amounts:

[0022] The ingredients include: 20-40 g of calcium-containing fermented lysate, 12-20 g of polyglycerol-10 laurate, 12-15 g of PEG-40 hydrogenated castor oil, 6-10 g of dioctyl carbonate, 5-7 g of butylene glycol, 5-7 g of propylene glycol, 0.6-0.8 g of 1,2-hexanediol, 0.5-1 g of lecithin, 0.03-0.1 g of palmitoyl pentapeptide-4, 3-4 g of 1,2-pentanediol, and 3.87-29.15 g of water.

[0023] Furthermore, the anti-aging composition comprises the following components and their amounts:

[0024] The mixture contains 40 g of calcium-containing fermentation lysate, 20 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 6 g of dioctyl carbonate, 5 g of butylene glycol, 6 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.5 g of lecithin, 0.03 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 3.87 g of water.

[0025] This invention is the first to prepare a fermented lysate with a high calcium content, exceeding 740 mg / kg. Studies have shown that the calcium in the calcium-containing fermented lysate provided by this invention is intracellular calcium, which can promote sebum metabolism, enhance skin barrier function, stabilize cell membrane structure, maintain the integrity of the stratum corneum, improve skin barrier function, and enhance skin's moisturizing and defensive capabilities. Simultaneously, this intracellular calcium can also improve dry skin, reduce pigmentation, and alleviate inflammatory responses. Furthermore, this intracellular calcium plays a crucial role in the synthesis of collagen fibers, promoting wound healing, maintaining the integrity of skin cell structure, regulating skin moisture balance, improving skin elasticity, and enhancing skin firmness and elasticity, thus exhibiting a significant anti-aging effect.

[0026] Furthermore, the palmitoyl pentapeptide-4 provided by this invention can act on skin fibroblasts to synthesize, repair, and rebuild the extracellular matrix. Its interaction with calcium-containing fermented lysate can effectively regulate cell activity, activate genes related to extracellular matrix renewal and cell proliferation, and promote the synthesis of more collagen, hyaluronic acid, and other substances in the skin, thereby reducing wrinkles and increasing skin elasticity. Simultaneously, the synergistic effect of components such as polyglycerol-10 laurate, PEG-40 hydrogenated castor oil, and lecithin can effectively promote skin absorption and improve the stability of the composition, resulting in a significant anti-aging effect.

[0027] In summary, compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0028] This invention is the first to prepare a high-calcium fermentation lysate, with a calcium content >740 mg / kg. Experiments have shown that this calcium-containing fermentation lysate can significantly promote the relative gene expression levels of collagen and elastin, exhibiting significant anti-wrinkle effects. Simultaneously, this calcium-containing fermentation lysate can effectively inhibit elastase activity, demonstrating a significant firming effect. Furthermore, this invention applies the calcium-containing fermentation lysate to cosmetics. The interaction of the components in this calcium-containing fermentation lysate composition not only improves skin absorption but also effectively enhances the stability of the composition, thus facilitating the stable exertion of its anti-aging effects. This makes it a relatively ideal anti-aging cosmetic. Attached Figure Description

[0029] Figure 1The stability appearance of the anti-aging composition prepared in Example 6 at 0, 1, 3, and 12 months is shown. Detailed Implementation

[0030] The present invention will be further described below through specific embodiments, but this is not intended to limit the invention. Those skilled in the art can make various modifications or improvements based on the basic ideas of the invention, but as long as they do not depart from the basic ideas of the invention, they are all within the scope of the invention. The sake yeast provided by the present invention has the Latin name: *Saccharomyces sake*, accession number: cfcc 3575, product number: bio-33961, and is supplied by: Zhili Zhongte (Wuhan) Biotechnology Co., Ltd. The bovine bone peptone was purchased from Beijing Hongrun Baoshun Technology Co., Ltd., and the ordinary peptone (bacteriological peptone) was purchased from Guangdong Huankai Microbial Technology Co., Ltd.

[0031] Example 1: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0032] Step S1: Dissolve each culture medium component in pure water until clear, pour into an Erlenmeyer flask to 300 mL, place in a high temperature and high pressure sterilizer and sterilize at 121 ℃ for 18 min. After sterilization, place in a clean bench and inoculate with sake yeast. The inoculation amount of sake yeast is 10%. Then ferment in a constant temperature shaking shaker at 28 ℃ and 140 rpm for 22 h to obtain fermentation broth.

[0033] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 10,000 rpm for 30 min, collect the precipitate, and perform low-temperature microfluidic extraction and disruption on the precipitate at a temperature of 25 ℃ and a flow rate of 18 L / min for 45 min to obtain calcium-containing fermentation lysate.

[0034] The culture medium in step S1 consists of the following components:

[0035] Glucose 110 g / L, sodium dihydrogen phosphate 0.4 g / L, disodium hydrogen phosphate 0.2 g / L, magnesium sulfate 0.15 g / L, bovine bone peptone 8 g / L, and pure water 881.25 g / L.

[0036] Example 2: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0037] Step S1: Dissolve each culture medium component in pure water until clear, pour into an Erlenmeyer flask to 300 mL, place in a high temperature and high pressure sterilizer and sterilize at 121 ℃ for 20 min. After sterilization, place in a clean bench and inoculate with sake yeast. The inoculation amount of sake yeast is 8%. Then ferment in a constant temperature shaking shaker at 25 ℃ and 160 rpm for 18 h to obtain fermentation broth.

[0038] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 11,000 rpm for 35 min. Collect the precipitate and perform low-temperature microfluidic extraction and disruption on the precipitate at a temperature of 20 ℃ and a flow rate of 20 L / min for 35 min to obtain calcium-containing fermentation lysate.

[0039] The culture medium in step S1 consists of the following components:

[0040] Glucose 50 g / L, sodium dihydrogen phosphate 0.5 g / L, disodium hydrogen phosphate 0.6 g / L, magnesium sulfate 0.05 g / L, bovine bone peptone 16 g / L, water 932.85 g / L.

[0041] Example 3: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0042] Step S1: Dissolve each culture medium component in pure water until clear, pour into an Erlenmeyer flask to 300 mL, place in a high temperature and high pressure sterilizer and sterilize at 121 ℃ for 22 min. After sterilization, place in a clean bench and inoculate with sake yeast. The inoculation amount of sake yeast is 5%. Then ferment in a constant temperature shaking shaker at 30 ℃ and 120 rpm for 24 h to obtain fermentation broth.

[0043] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 12000 rpm for 25 min, collect the precipitate, and perform low-temperature microfluidic extraction and disruption on the precipitate at a temperature of 30 ℃ and a flow rate of 15 L / min for 50 min to obtain calcium-containing fermentation lysate.

[0044] The culture medium in step S1 consists of the following components:

[0045] Glucose 150 g / L, sodium dihydrogen phosphate 0.8 g / L, disodium hydrogen phosphate 0.1 g / L, magnesium sulfate 0.35 g / L, bovine bone peptone 1 g / L, water 847.75 g / L.

[0046] Example 4: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0047] Step S1: Dissolve each culture medium component in pure water until clear, pour into an Erlenmeyer flask to 300 mL, place in a high temperature and high pressure sterilizer and sterilize at 121 ℃ for 22 min. After sterilization, place in a clean bench and inoculate with sake yeast. The inoculation amount of sake yeast is 5%. Then ferment in a constant temperature shaking shaker at 30 ℃ and 120 rpm for 24 h to obtain fermentation broth.

[0048] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 12000 rpm for 25 min, collect the precipitate, and perform low-temperature microfluidic extraction and disruption on the precipitate at a temperature of 30 ℃ and a flow rate of 15 L / min for 50 min to obtain calcium-containing fermentation lysate.

[0049] The culture medium in step S1 consists of the following components:

[0050] Glucose 120 g / L, sodium dihydrogen phosphate 0.6 g / L, disodium hydrogen phosphate 0.3 g / L, magnesium sulfate 0.1 g / L, bovine bone peptone 9 g / L, water 870 g / L.

[0051] Example 5: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0052] Step S1: Dissolve each culture medium component in pure water until clear, pour into an Erlenmeyer flask to 300 mL, place in a high temperature and high pressure sterilizer and sterilize at 121 ℃ for 18 min. After sterilization, place in a clean bench and inoculate with sake yeast. The inoculation amount of sake yeast is 10%. Then ferment in a constant temperature shaking shaker at 28 ℃ and 140 rpm for 22 h to obtain fermentation broth.

[0053] Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 10,000 rpm for 30 min, collect the precipitate, and perform low-temperature microfluidic extraction and disruption on the precipitate at a temperature of 25 ℃ and a flow rate of 18 L / min for 45 min to obtain calcium-containing fermentation lysate.

[0054] The culture medium in step S1 consists of the following components:

[0055] Glucose 110 g / L, sodium dihydrogen phosphate 0.7 g / L, disodium hydrogen phosphate 0.2 g / L, magnesium sulfate 0.14 g / L, bovine bone peptone 5 g / L, water 883.96 g / L.

[0056] Example 6: A composition with anti-aging effects

[0057] The anti-aging composition comprises the following ingredients and their amounts:

[0058] The calcium-containing fermentation lysate consisted of 30 g of calcium-containing fermentation lysate, 15 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 7 g of dioctyl carbonate, 5 g of butylene glycol, 5 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.8 g of lecithin, 0.1 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 18.5 g of water. The calcium-containing fermentation lysate was prepared according to Example 1.

[0059] Preparation method:

[0060] Step I: Add polyglycerol-10 laurate, PEG-40 hydrogenated castor oil, lecithin, and dioctyl carbonate to a beaker and heat to dissolve. After cooling, add palmitoyl pentapeptide-4 and stir until homogeneous to obtain phase A.

[0061] Step II: Add water, 1,2-hexanediol, 1,2-pentanediol, butanediol, propylene glycol, and calcium-containing fermentation lysate to a beaker and stir until homogenized to obtain phase B.

[0062] Step III: Pour the A phase obtained in Step I into the B phase obtained in Step II, and homogenize to obtain the final product.

[0063] Example 7: A composition with anti-aging effects

[0064] The anti-aging composition comprises the following ingredients and their amounts:

[0065] The calcium-containing fermentation lysate consisted of 20 g of calcium-containing fermentation lysate, 12 g of polyglycerol-10 laurate, 12 g of PEG-40 hydrogenated castor oil, 10 g of dioctyl carbonate, 6 g of butylene glycol, 6 g of propylene glycol, 0.8 g of 1,2-hexanediol, 1.0 g of lecithin, 0.05 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 29.15 g of water. The calcium-containing fermentation lysate was prepared in Example 2.

[0066] The preparation method is similar to that in Example 6.

[0067] Example 8: A composition with anti-aging effects

[0068] The anti-aging composition comprises the following ingredients and their amounts:

[0069] The calcium-containing fermentation lysate consisted of 40 g of calcium-containing fermentation lysate, 20 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 6 g of dioctyl carbonate, 5 g of butylene glycol, 6 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.5 g of lecithin, 0.03 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 3.87 g of water. The calcium-containing fermentation lysate was prepared in Example 3.

[0070] The preparation method is similar to that in Example 6.

[0071] Example 9: A composition with anti-aging effects

[0072] The anti-aging composition comprises the following ingredients and their amounts:

[0073] The calcium-containing fermentation lysate consisted of 25g of calcium-containing fermentation lysate, 13g of polyglycerol-10 laurate, 12g of PEG-40 hydrogenated castor oil, 8g of dioctyl carbonate, 7g of butylene glycol, 5g of propylene glycol, 0.6g of 1,2-hexanediol, 0.6g of lecithin, 0.09g of palmitoyl pentapeptide-4, 3g of 1,2-pentanediol, and 25.71g of water. The calcium-containing fermentation lysate was prepared according to Example 4.

[0074] The preparation method is similar to that in Example 6.

[0075] Example 10: A composition with anti-aging effects

[0076] The anti-aging composition comprises the following ingredients and their amounts:

[0077] The calcium-containing fermentation lysate consisted of 35 g of calcium-containing fermentation lysate, 15 g of polyglycerol-10 laurate, 13 g of PEG-40 hydrogenated castor oil, 9 g of dioctyl carbonate, 7 g of butylene glycol, 7 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.8 g of lecithin, 0.07 g of palmitoyl pentapeptide-4, 4 g of 1,2-pentanediol, and 8.53 g of water. The calcium-containing fermentation lysate was prepared in Example 5.

[0078] The preparation method is similar to that in Example 6.

[0079] Comparative Example 1: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0080] The culture medium consists of the following components:

[0081] Glucose 110 g / L, sodium dihydrogen phosphate 0.4 g / L, disodium hydrogen phosphate 0.2 g / L, magnesium sulfate 0.15 g / L, ordinary peptone 8 g / L, pure water 881.25 g / L.

[0082] The preparation method is similar to that in Example 1.

[0083] Comparative Example 2: A method for preparing a calcium-containing fermented lysate with anti-aging effects.

[0084] The culture medium consists of the following components:

[0085] Glucose 110 g / L, sodium dihydrogen phosphate 0.4 g / L, disodium hydrogen phosphate 0.2 g / L, magnesium sulfate 0.15 g / L, ordinary peptone 8 g / L, pure water 881.25 g / L, calcium chloride 0.5 g / L.

[0086] The preparation method is similar to that in Example 1.

[0087] Comparative Example 3: A composition with anti-aging effects

[0088] The anti-aging composition comprises the following ingredients and their amounts:

[0089] The calcium-containing fermentation lysate consisted of 30.1 g of calcium-containing fermentation lysate, 15 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 7 g of dioctyl carbonate, 5 g of butylene glycol, 5 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.8 g of lecithin, 3 g of 1,2-pentanediol, and 18.5 g of water. The calcium-containing fermentation lysate was prepared in Example 1.

[0090] The preparation method is similar to that in Example 6.

[0091] The difference from Example 6 is that palmitoyl pentapeptide-4 is missing.

[0092] Comparative Example 4: A composition with anti-aging effects

[0093] The anti-aging composition comprises the following ingredients and their amounts:

[0094] The calcium-containing fermentation lysate consisted of 30 g of calcium-containing fermentation lysate, 15.8 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 7 g of dioctyl carbonate, 5 g of butylene glycol, 5 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.1 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 18.5 g of water. The calcium-containing fermentation lysate was prepared according to Example 1.

[0095] The preparation method is similar to that in Example 6.

[0096] The difference from Example 6 is that lecithin is missing.

[0097] Experimental Example 1: Calcium Content Test of Calcium-Containing Fermentation Lysate

[0098] 1. Experimental Method:

[0099] The determination was carried out in accordance with GB 5009.268-2016 "National Food Safety Standard - Determination of Multiple Elements in Food". Specifically, the calcium concentration of the calcium-containing fermentation lysates prepared in Examples 1, 2, 3, 4, and 5 was determined by instrumental ICP-MS, and the detected calcium concentration was 1.00 mg / kg.

[0100] 2. Experimental Results:

[0101] The experimental results are shown in Table 1.

[0102] Group Calcium concentration (mg / kg) Example 1 987.3 Example 2 751.5 Example 3 901.3 Example 4 936.2 Example 5 969.4 Comparative Example 1 15.9 Comparative Example 2 67.6

[0103] 1. Experimental Method:

[0104] The anti-wrinkle efficacy of the compositions with anti-aging effects prepared in Examples 6, 7, 8, 9, 10, Comparative Example 3, and Comparative Example 4 was tested. Specifically, zebrafish embryos were used as a model, and real-time quantitative PCR was used for determination. The concentration of the test sample of the composition with anti-aging effect was 5% (mass concentration). The relative gene expression levels of cellular collagen and elastin were detected to comprehensively evaluate the anti-wrinkle efficacy of the samples.

[0105] 2. Experimental Results:

[0106] The experimental results are shown in Table 2.

[0107] Table 2. Test results of anti-wrinkle efficacy of compositions with anti-aging effects.

[0108] Example 6 Example 7 Example 8 Example 9 Example 10 Comparative Example 3 Comparative Example 4 col1a1 promotion rate 138% 105% 121% 91% 124% 87% 40% col1a1 promotion rate p-value 0.000098 0.00064 0.00021 0.00053 0.00074 0.000025 0.000076 col1a1b promotion rate 277% 209% 235% 195% 256% 72% 51% col1a1b promotion rate p-value 0.000028 0.00012 0.0075 0.0061 0.0017 0.00014 0.000024 col1a2 promotion rate 135% 119% 107% 115% 129% 65% 43% col1a2 promotion rate p-value 0.000011 0.00024 0.000019 0.00034 0.00040 0.00055 0.000010 Elastin promotion rate 221% 213% 204% 217% 203% 197% 63% Elastin promotion rate p-value 0.003 0.00037 0.00047 0.00053 0.00029 0.00032 0.000082

[0109] As shown in Table 2, the anti-aging composition prepared by the present invention has significant anti-wrinkle effects.

[0110] Experimental methods:

[0111] The inhibitory effect of the anti-aging composition prepared in Example 6 on elastase was tested. According to GDCTC / OT-L015 "Cosmetics - Elastase Inhibition Test Procedure", 0.1 mL of tris(hydroxymethyl)aminomethane buffer, 50 μL of elastase solution, and 50 μL of sample solution were added to a 96-well microplate. The plate was incubated at 25 °C for 20 min. MAAPVN (N-methoxysuccinyl-Ala-Ala-Pro-Val p-nitroaniline) and 50 μL of substrate solution were added. The total reaction volume was then increased to 250 μL with tris(hydroxymethyl)aminomethane buffer. The plate was incubated at 25 °C for another 60 min for color development. The elastase inhibition rate was measured at 405 nm using a microplate reader. The positive control was sodium selenopropyl, and the negative control was 0% sample solution.

[0112] 2. Experimental Results:

[0113] The experimental results are shown in Table 3.

[0114] Table 3. Inhibition results of elastase by compositions with anti-aging effects.

[0115]

[0116] As shown in Table 3, in the experiment, the elastase inhibition rate of the sample at the tested concentration of 3% was 30.7%, which was significantly different from the negative control (P < 0.05). This indicates that the anti-aging composition provided by the present invention has a firming effect.

[0117] Experimental methods:

[0118] A model of neutrophil aggregation induced by damage to neurothalamic cells in the lateral line region of zebrafish embryos using copper sulfate was tested. Twenty-four zebrafish embryos were exposed to 0.16 mg / L copper sulfate pentahydrate and 0.899 g / L of the sample solution from Example 6, with a positive control group (0.0036 mg / L indomethacin). After 40 min of exposure, the embryos were fixed and stained with Sudan Black. The number of neutrophils in the lateral line region was counted and statistically analyzed.

[0119] Experimental results:

[0120] The experimental results are shown in Table 4.

[0121] Table 4. Soothing effects of compositions with anti-aging effects

[0122] test Test concentration result Evaluation of soothing effects Example 6 0.899g / L 84.15%（p=0.00000001） Significant Positive control group 0.0036 mg / L 45.9%（p=0.0003044712） Significant

[0123] As shown in Table 4, in the experiment, the sample of Example 6, at a test concentration of 0.899 g / L, exhibited an inhibition rate of 84.15% for neutrophil aggregation in zebrafish embryos (p=0.00000001), which was significantly different from the positive control (P<0.05). This indicates that the anti-aging composition provided by the present invention can significantly inhibit neutrophil aggregation in zebrafish embryos and has a soothing effect.

[0124] 1. Experimental Method:

[0125] The anti-aging composition prepared in Example 6 was placed in a sealed container and left at room temperature for 12 months. The appearance changes of the test sample were observed at 0, 1, 3 and 12 months.

[0126] 2. Experimental Results:

[0127] Experimental results are as follows Figure 1 As shown in Table 5.

[0128] 2.1. Appearance of the anti-aging composition prepared in Example 6 after being placed at room temperature for 0, 1, 3, and 12 months, wherein: A--month 0, B--month 1, C--month 3, D--month 12.

[0129] 2.2 The appearance evaluation results of the anti-aging composition prepared in Example 6 after being left at room temperature for 0, 1, 3 and 12 months are shown in Table 5.

[0130] Table 5. Stability results of compositions with anti-aging effects

[0131] Month 0 Month 1 3rd month 12th month Example 6 A clear, yellow, transparent liquid with no sediment. A clear, yellow, transparent liquid with no sediment. A clear, yellow, transparent liquid with no sediment. A clear, yellow, transparent liquid with no sediment.

[0132] Depend on Figure 1 As shown in Table 5, the anti-aging composition provided by the present invention has high stability.

[0133] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A composition having an anti-aging efficacy, characterized in that, It contains the following ingredients and their amounts: The mixture contains 20-40 g of calcium-containing fermentation lysate, 12-20 g of polyglycerol-10 laurate, 12-15 g of PEG-40 hydrogenated castor oil, 6-10 g of dioctyl carbonate, 5-7 g of butylene glycol, 5-7 g of propylene glycol, 0.6-0.8 g of 1,2-hexanediol, 0.5-1 g of lecithin, 0.03-0.1 g of palmitoyl pentapeptide-4, 3-4 g of 1,2-pentanediol, and 3.87-29.15 g of water. The method for preparing the calcium-containing fermentation lysate is as follows: Step S1: Dissolve each culture medium component in pure water, sterilize, and then place it in a clean bench to inoculate the bacterial culture. Then ferment it in a constant temperature shaking shaker at a temperature of 25~30 ℃ and a speed of 120~160 rpm for 18~24 h to obtain the fermentation broth. Step S2: Centrifuge the fermentation broth obtained in step S1 at a speed of 10,000 to 12,000 rpm for 25 to 35 minutes. Take the precipitate and extract and break it using low-temperature microfluidic extraction to obtain calcium-containing fermentation lysate. The culture medium in step S1 consists of the following components: Glucose 90~150 g / L, sodium dihydrogen phosphate 0.4~0.8 g / L, disodium hydrogen phosphate 0.1~0.6 g / L, magnesium sulfate 0.05~0.4 g / L, bovine bone peptone 1~16 g / L, water 800~950 g / L; The strain used in step S1 is sake yeast, and the inoculation amount of sake yeast is 5-10%. The conditions for low-temperature microfluidic extraction and fragmentation in step S2 are: extraction for 30-50 min at a temperature of 20-30 ℃ and a flow rate of 15-20 L / min.

2. The composition with anti-aging effects as described in claim 1, characterized in that, It consists of the following components and their amounts: The mixture contains 40 g of calcium-containing fermentation lysate, 20 g of polyglycerol-10 laurate, 15 g of PEG-40 hydrogenated castor oil, 6 g of dioctyl carbonate, 5 g of butylene glycol, 6 g of propylene glycol, 0.6 g of 1,2-hexanediol, 0.5 g of lecithin, 0.03 g of palmitoyl pentapeptide-4, 3 g of 1,2-pentanediol, and 3.87 g of water.

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