Anti-aging composition suitable for sensitive skin as well as preparation method and application of anti-aging composition

Abstract

The invention discloses an anti-aging composition suitable for sensitive skin as well as a preparation method and application of the anti-aging composition, and belongs to the technical field of cosmetics. The composition takes a lactobacillus fermentation extract as a core component, and is matched with 1, 2-pentanediol, 1, 3-propylene glycol and capryloyl hydroximic acid. The lactobacillus fermentation extract is prepared by step-by-step extraction of tricholoma matsutake and rhizoma bletillae, lactobacillus fermentation, compound enzyme ultrasonic enzymolysis and centrifugal purification. The composition utilizes the synergistic effect of tricholoma matsutake and rhizoma bletillae, combines the micromolecule conversion effect of microbial fermentation and compound enzymatic hydrolysis to release active ingredients, has the effects of efficiently resisting oxidation, promoting collagen synthesis, repairing skin barrier and relieving and resisting inflammation, is mild in property and free of irritation, meets the anti-aging requirement of sensitive skin, and has a good application prospect. The skin care product can be widely applied to various skin care products such as water aqua, emulsion, essence, mask liquid and the like.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to an anti-aging composition suitable for sensitive skin, its preparation method and application, and particularly to an anti-aging composition based on Matsutake mushroom and Bletilla striata extracts obtained by fermentation with Lactobacillus. Background Technology

[0002] Aging is essentially a complex, multidimensional biological process involving progressive functional decline at the molecular, cellular, tissue, and system levels. From a core mechanism perspective, the essence of aging can be summarized as an imbalance between "damage accumulation and repair capacity." Over time, molecular damage within cells gradually exceeds the body's repair capacity, leading to abnormal cell function, tissue and organ degeneration, and ultimately triggering the overall aging phenotype.

[0003] However, the proportion of people with sensitive skin is increasing globally. These individuals typically have a weaker or damaged skin barrier, making them less resistant to external stimuli and prone to discomfort such as redness, stinging, itching, and tightness. Many highly effective anti-aging active ingredients, such as high concentrations of retinol, are often highly irritating and can easily trigger intolerance reactions in people with sensitive skin, thus limiting their application. Therefore, developing products that combine highly effective anti-aging properties with gentle, non-irritating ingredients, and even the ability to repair the skin barrier, has become a pressing technical challenge for the current cosmetics market.

[0004] In recent years, consumers have increasingly favored natural and safe cosmetic ingredients. Matsutake mushroom (Tricholoma matsutake), a precious edible and medicinal fungus, has extracts rich in polysaccharides, peptides, amino acids, and other bioactive substances. Studies have shown that matsutake polysaccharides possess excellent antioxidant and moisturizing capabilities. Bletilla striata, a traditional Chinese medicine, has a main active ingredient, Bletilla striata polysaccharide (BSP), which has been proven to promote wound healing, reduce inflammation, moisturize, and repair the skin barrier.

[0005] Furthermore, microbial fermentation technology is increasingly being applied in the cosmetics industry. Fermenting plant substrates with specific microorganisms (such as lactobacillus) not only breaks down large molecules into smaller molecules that are more easily absorbed by the skin, but also produces various beneficial metabolites (i.e., post-fermenters) such as lactic acid, short-chain fatty acids, and prebiotics during the fermentation process. This enhances the efficacy of the raw materials and reduces their potential irritation. Fermentation products have been shown to improve the skin's microecology and strengthen the skin barrier function, making them particularly suitable for sensitive skin.

[0006] While studies have reported on the individual applications of matsutake mushrooms or Bletilla striata, research on applying both through co-fermentation with Lactobacillus and combined with specific enzymatic hydrolysis technology to an anti-aging composition for sensitive skin has not been reported. Therefore, this invention provides an anti-aging composition suitable for sensitive skin that synergistically leverages the antioxidant and anti-aging effects of matsutake mushrooms and the barrier-repairing and soothing effects of Bletilla striata. Furthermore, microbial fermentation technology enhances its overall efficacy, gentleness, and skin absorption rate, thus providing a safe and effective anti-aging solution for people with sensitive skin. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the prior art and provide an anti-aging composition suitable for sensitive skin. This composition has good antioxidant, anti-aging and soothing anti-inflammatory effects, while being mild and not causing skin irritation.

[0008] This invention provides an anti-aging composition suitable for sensitive skin, comprising the following raw materials in parts by weight: 80-95 parts of Lactobacillus fermentation extract, 3-4 parts of 1,2-pentanediol, 5-9 parts of 1,3-propanediol, and 0.05-0.3 parts of capryloyl hydroxamic acid.

[0009] The preparation method of the Lactobacillus fermentation extract includes the following steps: S1. After pulverizing the dried matsutake mushrooms, add water at a material-to-liquid ratio of 1:(10-30), extract at 80-100℃ for 1-4 hours, and filter to obtain matsutake mushroom extract; S2. Add Bletilla striata powder to 30-50% ethanol solution at a material-to-liquid ratio of 1:(15-40), and extract by ultrasonication at 50-80℃ for 1-3 hours. Filter to obtain Bletilla striata extract. S3. Mix the matsutake mushroom extract and bletilla striata extract at a mass ratio of (3-8):1 to obtain a mixed extract. Add a nitrogen source and inorganic salt to the mixed extract, adjust the pH to 5.0-7.0, sterilize for the first time, inoculate with lactic acid bacteria, ferment at 30-42℃ for 24-120 hours, filter, sterilize for the second time, and obtain the fermentation broth. S4. Add the compound enzyme to the fermentation broth from step S3, sonicate, centrifuge, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0010] Preferably, in step S1, the pulverized matsutake mushroom has a particle size of 50-100 mesh; in step S2, the powdered Bletilla striata has a particle size of 40-80 mesh.

[0011] Preferably, the inorganic salt in step S3 is one or more of ammonium chloride, potassium dihydrogen phosphate, sodium sulfate, and calcium chloride, and the amount added is 0.5 to 3 wt% of the mass of the mixed extract; the nitrogen source is one or more of peptone, yeast extract, or soybean protein, and the amount added is 3 to 10 wt% of the mass of the mixed extract.

[0012] Preferably, the lactobacillus in step S3 is one or more of Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus acidophilus, and the inoculation amount is 1 to 10% of the mass of the mixed extract.

[0013] Preferably, the concentration of the lactobacillus is 1×10⁻⁶. 8 ~1×10 10 CFU / mL.

[0014] Preferably, the complex enzyme in step S3 is a mixture of papain and cellulase in a mass ratio of 5-10:1-4.

[0015] Preferably, the filtration in step S3 is performed using a 0.5μm to 2.5μm polypropylene microfiltration membrane; both the first and second sterilizations are low-temperature pasteurizations, with a sterilization temperature of 80 to 100°C and a sterilization time of 30 to 60 minutes.

[0016] Preferably, the amount of the compound enzyme added in step S4 relative to the fermentation broth is 0.1~0.5 wt%, the power of the ultrasound is 500~800W, the reaction temperature is 35~45℃, and the reaction time is 1~2 hours.

[0017] Preferably, the anti-aging composition of the present invention may further include 0.1 to 1.0 parts of a penetrant, said penetrant including at least one selected from ceramide, phytosterol, and low molecular weight hyaluronic acid. More preferably, the penetrant is a combination of 0.1 to 0.5 parts of ceramide, 0.1 to 1.0 parts of phytosterol, and 0.1 to 0.8 parts of low molecular weight hyaluronic acid.

[0018] Preferably, the low molecular weight hyaluronic acid has a molecular weight of 5~20kDa.

[0019] To further improve the skin permeability of active ingredients in fermented extracts (such as matsutake polysaccharides and Bletilla striata polysaccharides), this invention further adds a composite penetration-enhancing system consisting of 0.05-0.3 wt% ceramides, 0.1-0.5 wt% phytosterols, and 0.1-0.8 wt% low molecular weight hyaluronic acid to the anti-aging composition. This system, on the one hand, reduces permeability resistance by replenishing stratum corneum lipids and repairing the skin barrier; on the other hand, it promotes the penetration of active ingredients by enhancing stratum corneum hydration and forming permeation channels. All penetration-enhancing ingredients are of natural origin or are inherent skin components, gentle and non-irritating, highly compatible with the needs of sensitive skin, and have not caused skin erythema, stinging, or other irritation reactions.

[0020] This invention innovatively utilizes a co-fermentation technology combining matsutake mushroom extract and Bletilla striata extract, overcoming the limitations of single-component formulations, achieving complementary advantages, and enhancing the overall efficacy of the product. This is mainly reflected in the following four aspects: First, it enhances antioxidant capacity. The polysaccharides in Bletilla striata and the polyphenols in matsutake mushrooms have phenolic hydroxyl groups, which can directly neutralize ROS. The glutathione and superoxide dismutase (SOD) analogs produced by lactobacillus metabolism further enhance the efficiency of free radical scavenging.

[0021] Secondly, it promotes collagen synthesis. Bletilla striata oligosaccharides (after fermentation) can activate the TGF-β / Smad signaling pathway in fibroblasts and promote the expression of type I and type III collagen genes. Adenosine analogues produced by matsutake fermentation can enhance fibroblast proliferation and improve collagen synthesis efficiency.

[0022] Third, it repairs the skin barrier. The amino acids produced by fermentation are the core components of the natural moisturizing factor (NMF) in the stratum corneum, which can increase the moisture content of the stratum corneum; Bletilla striata oligosaccharides can promote the secretion of ceramides (key lipids of the skin barrier) by keratinocytes, thereby enhancing the density of the stratum corneum.

[0023] Fourth, it inhibits the release of inflammatory factors. Terpenoid derivatives produced by matsutake fermentation can block the release of pro-inflammatory factors such as IL-6 and TNF-α from macrophages and keratinocytes; cell wall components of lactobacillus can activate the anti-inflammatory phenotype of skin immune cells and balance the immune response.

[0024] The present invention selects matsutake extract because the fermentation of matsutake extract enhances its ability to resist mitochondrial dysfunction: small molecules after matsutake fermentation can be transported into cells and mitochondria through the transporter OCTN-1 in keratinocytes and fibroblasts, directly scavenging reactive oxygen free radicals and exerting antioxidant and cell protection effects.

[0025] The fermentation of matsutake mushroom extract achieves multidimensional improvement in mitochondrial disorders through the "transformation" and "addition" effects of microbial metabolism: on the one hand, fermentation optimizes the original active ingredients of matsutake (such as smaller molecules and enhanced activity), improving its antioxidant and energy regulation capabilities; on the other hand, the newly added metabolites (such as short-chain fatty acids, peptides, and coenzyme precursors) synergistically enhance the ability to resist mitochondrial disorders through mechanisms such as structural protection, signaling pathway activation, and repair promotion. The fermentation of Bletilla striata extract produces small-molecule Bletilla striata polysaccharides. Different molecular weights of Bletilla striata polysaccharides exhibit targeted differences in their effects, with multi-level functional complementarity, ultimately achieving comprehensive regulation of the skin barrier and keratinocyte health.

[0026] This invention utilizes the mitochondrial activation properties (qi-tonifying) of matsutake mushrooms and the extracellular matrix remodeling ability (tissue regeneration) of Bletilla striata to achieve precise coupling of energy and structure at the subcellular level through microbial fermentation, thereby achieving: fibroblast activity reaching physiological peak state and skin barrier thickness restored to youthful baseline.

[0027] This invention involves adding a complex enzyme of papain and cellulase to the fermentation broth after Lactobacillus fermentation. Its core function is based on the specific catalytic and synergistic effects of the enzymes: papain specifically hydrolyzes residual allergenic macromolecular plant proteins (such as leukoglobulin) in the fermentation broth into small peptides or amino acids without allergenic risk; cellulase, on the other hand, degrades the residual cellulose structure in the plant cell wall stepwise, releasing encapsulated polysaccharides, flavonoids, and other anti-aging active ingredients. The synergistic effect of these two enzymes not only improves the protein degradation rate and the release efficiency of active ingredients but also further converts some macromolecular polysaccharides into easily permeable oligosaccharides. Simultaneously, it removes undegraded impurities to optimize the stability of the fermentation broth system. Furthermore, the small peptides and amino acids in the enzymatic hydrolysis products have anti-inflammatory and soothing effects, thus achieving a dual optimization of efficacy and stability while ensuring safe tolerance for sensitive skin.

[0028] The present invention also provides a method for preparing an anti-aging composition suitable for sensitive skin, specifically including the following steps: stirring each component evenly, adjusting the pH to 6.5-7, to obtain an anti-aging composition suitable for sensitive skin.

[0029] Preferably, the present invention also provides the use of the aforementioned anti-aging composition suitable for sensitive skin in the preparation of skin care products.

[0030] Preferably, the present invention also provides a skin care product containing 1-100% of the anti-aging composition suitable for sensitive skin described above.

[0031] Preferably, the skin care product formulation is selected from at least one of aqueous solution, lotion, serum, or facial mask liquid.

[0032] The advantages or beneficial effects of the anti-aging composition of the present invention suitable for sensitive skin include at least the following: This invention utilizes the co-fermentation of matsutake mushroom and Bletilla striata extracts with Lactobacillus, combined with a complex enzymatic hydrolysis process using papain and cellulase. This achieves a synergistic complementarity between the antioxidant and anti-aging effects of matsutake mushroom and the barrier repair and soothing effects of Bletilla striata. Fermentation and enzymatic hydrolysis transform large molecules into smaller, more easily absorbed active substances, significantly improving the product's gentleness and efficacy while eliminating potential allergens. The addition of a complex penetration-enhancing system composed of ceramides, phytosterols, and low-molecular-weight hyaluronic acid further enhances the permeability of the active ingredients. All ingredients are naturally derived or are inherent to the skin, ensuring gentleness and non-irritation. The composition of this invention exerts its anti-aging effect through multiple targets, including strengthening antioxidant capacity, promoting collagen synthesis, repairing the skin barrier, and inhibiting the release of inflammatory factors, making it suitable for the specific needs of sensitive skin and various skincare product formulations. Attached Figure Description

[0033] Figure 1 The effect of the composition for anti-aging of sensitive skin in Example 1 on the expression of moisturizing-related genes in HaCaT cells.

[0034] Figure 2 The image below shows the effect of the anti-aging composition for sensitive skin in Example 1 on cell migration ability.

[0035] Figure 3 The effect of the composition for sensitive skin anti-aging in Example 1 on the migration ability of HaCaT cells.

[0036] Figure 4 The effect of the composition for sensitive skin anti-aging in Example 1 on collagen gene expression in photoaged HSF cells. Detailed Implementation

[0037] To more clearly illustrate the purpose, technical solution, and advantages of this invention, the technical solution of this invention will be described in detail below through specific embodiments. It should be noted that these embodiments are only for illustrating this invention and not for limiting its scope of protection; the actual scope of protection of this invention should be determined by the claims.

[0038] Unless otherwise specified, the materials and reagents used in the following examples and comparative examples are commercially available. Unless otherwise specified, the amount of each component in the following examples is 1 g per part by weight.

[0039] I. Experimental Materials: Lactobacillus plantarum, accession number: CCTCC AB 2013128, purchased from China Center for Type Culture Collection.

[0040] Lactobacillus paracasei, accession number: CCTCC CB 20081797, purchased from China Center for Type Culture Collection.

[0041] Lactobacillus acidophilus, accession number: CGMCC 1.12735, was purchased from the China General Microbiological Culture Collection Center. All other materials were commercially available products.

[0042] II. Examples and Comparative Examples Example 1 (1) Preparation of Lactobacillus fermentation extract: S1. Take 500g of dried matsutake mushrooms, grind them to 80 mesh, add 10L of water at a material-to-liquid ratio of 1:20, extract at 90℃ for 2 hours, and filter to obtain matsutake mushroom extract.

[0043] S2. Take 100g of Bletilla striata powder, grind it to 60 mesh, add 2.5L of 40% ethanol solution at a material-to-liquid ratio of 1:25, and extract it by ultrasonication at 70℃ for 2 hours. Filter to obtain Bletilla striata extract.

[0044] S3. Mix the matsutake mushroom extract and the bletilla striata extract at a mass ratio of 5:1 to obtain 6 kg of mixed extract; add 180 g of peptone and 30 g of ammonium chloride as nitrogen source and inorganic salt; adjust the pH to 6.3; pasteurize (90℃, 30 minutes); inoculate with Lactobacillus plantarum (concentration 1×10⁻⁶). 9 (CFU / mL, inoculum 5%), fermented at 37°C for 48 hours; filtered using a 1.5μm polypropylene microfiltration membrane; pasteurized again (90°C, 40 minutes) to obtain the fermentation broth.

[0045] S4. Add a mixture of papain and cellulase in a mass ratio of 8:1 to the fermentation broth, with a total addition of 18g; control the ultrasonic power at 600W, the reaction temperature at 37℃, and the reaction time at 1.5 hours; after enzymatic hydrolysis, pasteurize (90℃, 30 minutes); centrifuge at 5000r / min for 20 minutes, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0046] (2) Preparation of anti-aging composition: 90 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 6 parts of 1,3-propanediol, and 0.2 parts of capryloyl hydroxamic acid were stirred evenly and the pH was adjusted to 6.5 to obtain an anti-aging composition suitable for sensitive skin.

[0047] Example 2 (1) Preparation of Lactobacillus fermentation extract: S1. Take 600g of dried matsutake mushrooms, grind them to 70 mesh, add 15L of water at a material-to-liquid ratio of 1:25, extract at 85℃ for 3 hours, and filter to obtain matsutake mushroom extract.

[0048] S2. Take 120g of Bletilla striata powder, grind it to 50 mesh, add 3.6L of 35% ethanol solution at a material-to-liquid ratio of 1:30, and extract it by ultrasonication at 65℃ for 1.5 hours. Filter to obtain Bletilla striata extract.

[0049] S3. Mix the matsutake mushroom extract and the Bletilla striata extract at a mass ratio of 6:1 to obtain 5 kg of mixed extract; add 250 g of yeast extract powder and 75 g of potassium dihydrogen phosphate as nitrogen source and inorganic salt; adjust the pH to 6.0; pasteurize (88℃, 40 minutes); inoculate with Lactobacillus paracasei (concentration 5 × 10⁻⁶). 9 (CFU / mL, inoculum 6%), fermented at 35°C for 72 hours; filtered using a 1.2μm polypropylene microfiltration membrane; pasteurized again (88°C, 35 minutes) to obtain the fermentation broth.

[0050] S4. Add a mixture of papain and cellulase in a mass ratio of 7:2 to the fermentation broth, with a total addition of 15g (0.3wt% of the fermentation broth mass); control the ultrasonic power at 700W, the reaction temperature at 40℃, and the reaction time at 1.2 hours; after enzymatic hydrolysis, pasteurize (88℃, 30 minutes); centrifuge at 5500r / min for 20 minutes, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0051] (2) Preparation of anti-aging composition: 85 parts of Lactobacillus fermentation extract, 3.5 parts of 1,2-pentanediol, 7 parts of 1,3-propanediol, and 0.15 parts of capryloyl hydroxamic acid were stirred evenly and the pH was adjusted to 6.7 to obtain an anti-aging composition suitable for sensitive skin.

[0052] Example 3 (1) Preparation of Lactobacillus fermentation extract: S1. Take 400g of dried matsutake mushrooms, grind them to 50 mesh, add 6L of water at a material-to-liquid ratio of 1:15, extract at 95℃ for 1.5 hours, and filter to obtain matsutake mushroom extract.

[0053] S2. Take 80g of Bletilla striata powder, grind it to 40 mesh, add 1.6L of 50% ethanol solution at a material-to-liquid ratio of 1:20, and extract it by ultrasonication at 55℃ for 2.5 hours. Filter to obtain Bletilla striata extract.

[0054] S3. Mix the matsutake mushroom extract and the Bletilla striata extract at a mass ratio of 8:1 to obtain 4.8 kg of mixed extract; add 384 g of soybean protein (8 wt% of the mixed extract mass) and 96 g of sodium sulfate (2 wt% of the mixed extract mass) as nitrogen source and inorganic salt; adjust the pH to 6.0; pasteurize (95℃, 25 minutes); inoculate with Lactobacillus acidophilus (concentration 1×10⁻⁶). 10 (CFU / mL, inoculum 8%), fermented at 39°C for 96 hours; filtered using a 2.0 μm polypropylene microfiltration membrane; pasteurized again (95°C, 25 minutes) to obtain the fermentation broth.

[0055] S4. Add a mixture of papain and cellulase in a mass ratio of 10:3 to the fermentation broth, with a total addition of 19.2g (0.4wt% of the fermentation broth mass); control the ultrasonic power at 500W, the reaction temperature at 42℃, and the reaction time at 2 hours; after enzymatic hydrolysis, pasteurize (95℃, 25 minutes); centrifuge at 5000r / min for 20 minutes, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0056] (2) Preparation of anti-aging composition: 88 parts of Lactobacillus fermentation extract, 3.8 parts of 1,2-pentanediol, 8 parts of 1,3-propanediol, and 0.2 parts of capryloyl hydroxamic acid were stirred evenly and the pH was adjusted to 6.6 to obtain an anti-aging composition suitable for sensitive skin.

[0057] Example 4 (1) Preparation of Lactobacillus fermentation extract: S1. Take 700g of dried matsutake mushrooms, grind them to 100 mesh, add 21L of water at a material-to-liquid ratio of 1:30, extract at 80℃ for 4 hours, and filter to obtain matsutake mushroom extract.

[0058] S2. Take 140g of Bletilla striata powder, grind it to 80 mesh, add 5.6L of 30% ethanol solution at a material-to-liquid ratio of 1:40, and extract it by ultrasonication at 80℃ for 1 hour. Filter to obtain Bletilla striata extract.

[0059] S3. Mix the matsutake mushroom extract and the Bletilla striata extract at a mass ratio of 3:1 to obtain 7.2 kg of mixed extract; add 216 g of peptone-yeast extract mixture (3:2, 3 wt% of the mixed extract mass) and 144 g of potassium dihydrogen phosphate-calcium chloride mixture (1:1, 2 wt% of the mixed extract mass) as nitrogen source and inorganic salt; adjust the pH to 6.0; pasteurize (85℃, 50 minutes); inoculate with Lactobacillus plantarum (concentration 2×10⁻⁶). 9 (CFU / mL, inoculum 3%), fermented at 38°C for 120 hours; filtered using a 0.8μm polypropylene microfiltration membrane; pasteurized again (85°C, 50 minutes) to obtain the fermentation broth.

[0060] S4. Add a mixture of papain and cellulase in a mass ratio of 5:1 to the fermentation broth, with a total addition of 7.2g (0.1wt% of the fermentation broth mass); control the ultrasonic power at 800W, the reaction temperature at 35℃, and the reaction time at 1.8 hours; after enzymatic hydrolysis, pasteurize (85℃, 30 minutes); centrifuge at 4500r / min for 20 minutes, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0061] (2) Preparation of anti-aging composition: 95 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 5 parts of 1,3-propanediol, and 0.05 parts of capryloyl hydroxamic acid were stirred evenly and the pH was adjusted to 6.8 to obtain an anti-aging composition suitable for sensitive skin.

[0062] Example 5 (1) Preparation of Lactobacillus fermentation extract: S1. Take 550g of dried matsutake mushrooms, grind them to 90 mesh, add 12.1L of water at a material-to-liquid ratio of 1:22, extract at 92℃ for 2.5 hours, and filter to obtain matsutake extract.

[0063] S2. Take 110g of Bletilla striata powder, grind it to 70 mesh, add 3.08L of 45% ethanol solution at a material-to-liquid ratio of 1:28, and extract it by ultrasonication at 68℃ for 1.8 hours. Filter to obtain Bletilla striata extract.

[0064] S3. Mix the matsutake mushroom extract and the Bletilla striata extract at a mass ratio of 4:1 to obtain 6.5 kg of mixed extract; add 390 g of yeast extract powder-soybean protein mixture (2:1, 6 wt% of the mixed extract mass) and 97.5 g of ammonium chloride-sodium sulfate mixture (2:1, 1.5 wt% of the mixed extract mass) as nitrogen source and inorganic salt; adjust the pH to 5.8; pasteurize (92℃, 35 minutes); inoculate with Lactobacillus paracasei (concentration 8 × 10⁻⁶). 9 (CFU / mL, inoculum 10%), fermented at 36°C for 60 hours; filtered using a 2.5μm polypropylene microfiltration membrane; pasteurized again (92°C, 35 minutes) to obtain the fermentation broth.

[0065] S4. Add a mixture of papain and cellulase in a mass ratio of 9:4 to the fermentation broth, with a total addition of 32.5g (0.5wt% of the fermentation broth mass); control the ultrasonic power at 650W, the reaction temperature at 45℃, and the reaction time at 1 hour; after enzymatic hydrolysis, perform pasteurization (92℃, 30 minutes); centrifuge at 5000r / min for 20 minutes, collect the supernatant, and obtain the Lactobacillus fermentation extract.

[0066] (2) Preparation of anti-aging composition: 92 parts of Lactobacillus fermentation extract, 4 parts of 1,2-pentanediol, 9 parts of 1,3-propanediol, and 0.3 parts of capryloyl hydroxamic acid were stirred evenly and the pH was adjusted to 6.8 to obtain an anti-aging composition suitable for sensitive skin.

[0067] Example 6 (1) The preparation of the Lactobacillus fermentation extract is the same as in Example 1.

[0068] (2) Preparation of anti-aging composition: 90 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 6 parts of 1,3-propanediol, 0.2 parts of capryloyl hydroxamic acid, and 0.5 parts of ceramide were stirred evenly and the pH was adjusted to 6.5 to obtain an anti-aging composition suitable for sensitive skin.

[0069] Example 7 (1) The preparation of the Lactobacillus fermentation extract is the same as in Example 1.

[0070] (2) Preparation of anti-aging composition: 90 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 6 parts of 1,3-propanediol, 0.2 parts of capryloyl hydroxamic acid, and 0.8 parts of phytosterol were mixed evenly and the pH was adjusted to 6.5 to obtain an anti-aging composition suitable for sensitive skin.

[0071] Example 8 (1) The preparation of the Lactobacillus fermentation extract is the same as in Example 1.

[0072] (2) Preparation of anti-aging composition: 90 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 6 parts of 1,3-propanediol, 0.2 parts of capryloyl hydroxamic acid, and 0.6 parts of low molecular weight hyaluronic acid (0.6 parts, molecular weight 20kDa) were stirred evenly and the pH was adjusted to 6.5 to obtain an anti-aging composition suitable for sensitive skin.

[0073] Example 9 (1) The preparation of the Lactobacillus fermentation extract is the same as in Example 1.

[0074] (2) Preparation of anti-aging composition: 90 parts of Lactobacillus fermentation extract, 3 parts of 1,2-pentanediol, 6 parts of 1,3-propanediol, 0.2 parts of capryloyl hydroxamic acid, 0.5 parts of ceramide, 0.8 parts of phytosterol, and 0.6 parts of low molecular weight hyaluronic acid (molecular weight 20kDa) were stirred evenly and the pH was adjusted to 6.5 to obtain an anti-aging composition suitable for sensitive skin.

[0075] Comparative Example 1 Compared with Example 1, an equal amount of matsutake mushroom extract was used instead of Bletilla striata extract, and the other conditions were the same as in Example 1.

[0076] Comparative Example 2 Compared with Example 1, an equal amount of Bletilla striata extract was used instead of matsutake mushroom extract, and the other conditions were the same as in Example 1.

[0077] Comparative Example 3 Compared with Example 1, the enzymatic hydrolysis in step S4 is a single papain, and the other conditions are the same as in Example 1.

[0078] Comparative Example 4 Compared with Example 1, the enzymatic hydrolysis in step S4 is performed with a single cellulase, and the other conditions are the same as in Example 1.

[0079] Comparative Example 5 Compared with Example 1, the enzymatic hydrolysis process in step S4 was omitted, and the fermentation broth was used directly for the Lactobacillus fermentation extract. The remaining conditions were the same as in Example 1.

[0080] III. Performance Characterization 1. Antioxidant efficacy test DPPH free radical scavenging rate: Refer to GB / T 39199-2020 to determine the DPPH free radical scavenging ability of each composition.

[0081] 2. Anti-aging efficacy test Fibroblast collagen synthesis promotion rate: Using a human fibroblast (HFF-1) model, the content of type I collagen (COL1A1) was detected by ELISA, and the promotion rate was calculated.

[0082] Fibroblast proliferation rate: The effect of each composition on HFF-1 cell proliferation was detected by CCK-8 assay.

[0083] 3. Soothing and anti-inflammatory efficacy test The inhibition rate of IL-6 and TNF-α secretion by samples was detected using an LPS-induced HaCaT cell model. Specifically, RAW264.7 cells were collected by centrifugation and the cell density was adjusted to 2.5 × 10⁻⁶ cells / year. 5 The drug was seeded in 12-well plates at a concentration of 200 ng / mL and cultured overnight in a cell culture incubator. After RAW264.7 cells adhered, the drug was diluted with serum-free basal medium and the stimulant LPS was added to a final concentration of 200 ng / mL. After culturing for 6 h, RNA was extracted and the expression level of inflammatory factor mRNA in RAW264.7 cells was detected by qPCR.

[0084] 4. Skin barrier repair test Transdermal Water Loss (TEWL) Measurement: For the human transdermal water loss (TEWL) test, several volunteers with sensitive skin (meeting the CB / T 35916-2018 criteria for sensitive skin) were selected. The samples were used continuously for 7 days. Facial TEWL values ​​were measured before use (Day 0) and after 7 days of use. The TEWL reduction rate was calculated. TEWL reduction rate = (Initial TEWL value on Day 0 - Terminating TEWL value on Day 7) / Initial TEWL value on Day 0 × 100%. A higher TEWL reduction rate indicates a better effect on skin barrier repair.

[0085] Table 1: Performance test results of the examples and comparative examples are shown in the table below:

[0086] As shown in Table 1, the anti-aging compositions for sensitive skin in Examples 1-9, through co-fermentation of matsutake mushroom and Bletilla striata extracts with Lactobacillus and combined with a complex enzymatic hydrolysis process using papain and cellulase, significantly enhanced the antioxidant, anti-aging, and soothing anti-inflammatory effects of the products, while ensuring their mildness and safety. The compositions in Examples 1-9 were significantly superior to those in Comparative Examples 1-5 in terms of DPPH free radical scavenging rate, fibroblast proliferation rate, collagen synthesis, inflammatory factor inhibition rate, and TEWL reduction rate, demonstrating the synergistic effect of co-fermentation and the complex enzymatic hydrolysis process.

[0087] Comparing Examples 1 and 6-9, the anti-aging compositions of Examples 6-9 further incorporated ceramides, phytosterols, and low molecular weight hyaluronic acid as osmotic agents. This reduced osmotic resistance and enhanced the transport of active ingredients, facilitating improved stratum corneum hydration, the formation of osmotic channels to promote active ingredient penetration, and ultimately enhancing antioxidant, anti-aging, and soothing anti-inflammatory effects. Compared to the single penetration-enhancing system of Examples 6-8, Example 9 used a synergistic osmotic system formed by ceramides, phytosterols, and low molecular weight hyaluronic acid, which improved active ingredient penetration and significantly increased TNF-α and IL-6 inhibition rates.

[0088] Comparative Example 1 showed a significant decrease in the TEWL rate related to barrier repair and the inhibition rate of TNF-α and IL-6 related to anti-inflammation due to the lack of the activation effect of Bletilla striata oligosaccharides on fibroblast signaling pathways, resulting in insufficient collagen synthesis and cell proliferation. Comparative Example 2 lacked the key anti-aging effects of small molecule substances after matsutake fermentation in clearing ROS, protecting mitochondrial function, and promoting fibroblast proliferation with adenosine analogues, resulting in a significant decline in its antioxidant capacity and collagen synthesis promotion rate.

[0089] Comparative Example 3 used a single papain enzyme for hydrolysis, which could only hydrolyze large molecular proteins but could not degrade the cellulose remaining in the plant cell wall, resulting in insufficient release of the encapsulated polysaccharides, flavonoids, and other anti-aging active ingredients, and insufficient utilization of active ingredients. Comparative Example 4 used a single cellulase enzyme for hydrolysis, which could degrade cellulose and release some active ingredients, but could not hydrolyze large molecular plant proteins, posing a potential risk of irritation and affecting the contact efficiency between active ingredients and the skin. Comparative Example 5 directly omitted the enzymatic hydrolysis process, and the large molecular proteins, cellulose, and polysaccharides remaining in the fermentation broth were difficult to be absorbed by the skin. In addition, the large molecular proteins may cause discomfort to sensitive skin, and the undegraded impurities also affected the stability of the system.

[0090] 5. Efficacy characterization of anti-aging compositions for sensitive skin.

[0091] (1) Effects on the expression of moisturizing-related genes in HaCaT cells (I) Experimental conditions: Cell line: HaCaT cells Cell density: 15*10 4 / mL; 12-well plate Tested Sample: Example 1 - Anti-aging Raw Materials for Sensitive Skin Positive control: 2 mg / mL HA Detection method: qPCR (II) Experimental methods: HaCaT cells were collected and seeded in 12-well plates. After the cells adhered, the basal medium containing different concentrations of the test samples was replaced. After culturing for 48 h, RNA was extracted and the expression level of moisturizing-related genes in HaCaT cells was detected by qPCR.

[0092] *: P < 0.05 compared to the control group; **: P < 0.01 compared to the control group; ***: P < 0.001 compared to the control group; ***: P < 0.0001 compared to the control group.

[0093] Experimental results: Figure 1 The results showed that, compared with the control group, the concentration of the tested sample at 0.125 mg / mL significantly upregulated the expression of moisturizing-related genes AQP3 and FLG mRNA, and the concentration at 0.25 mg / mL significantly upregulated the expression of moisturizing-related gene FLG mRNA, indicating that the tested sample has moisturizing effect under this experimental model.

[0094] (2) Effects on HaCaT cell migration ability (I) Experimental conditions: Cell line: HaCaT cells Density: 3*10 5 / ml Tested Sample: Example 1 - Anti-aging Raw Materials for Sensitive Skin Positive control: 8% FBS (II) Experimental method: HaCaT cells in the logarithmic growth phase were seeded into 12-well plates. After 24 hours, when the cells had filled the bottom, the bottom was gently cut with a 200 μL pipette tip to simulate human skin injury. The culture medium was aspirated and the detached cells were gently rinsed with PBS. Then, culture medium containing different concentrations of the test sample was added. A blank control group and a positive control group were set up. Microscopic images were taken at 0 h, 12 h and 24 h. The scar area was then statistically analyzed and the healing rate was calculated using ImageJ software.

[0095] *: P < 0.05 compared to the control group; **: P < 0.01 compared to the control group; ***: P < 0.001 compared to the control group; ***: P < 0.0001 compared to the control group.

[0096] Results analysis: Figure 2-3The results showed that the cell scratch healing area of ​​the anti-aging raw material for sensitive skin was significantly increased at 12 h and 24 h after scratching, which was significantly different from the control group.

[0097] (3) Effect of the tested samples on collagen gene expression in photoaged HSF cells (I) Experimental conditions: Cell line: HSF cells Cell density: 6*10 4 / mL; 12-well plate Tested Sample: Example 1 - Anti-aging Raw Materials for Sensitive Skin Positive control: 5 μg / mL retinoic acid Detection method: qPCR (II) Experimental Methods: HSF cells were collected and seeded in 12-well plates. After cell adhesion, the cells were irradiated with UVB for 2 days, with a total dose of 400 mJ / cm². 2 Complete culture medium of different concentrations of the test samples was added, and after culturing for 48 h, RNA was extracted and the expression level of collagen gene in HSF cells was detected by qPCR.

[0098] *: P < 0.05 compared to the model group; **: P < 0.01 compared to the model group; ***: P < 0.001 compared to the model group; ***: P < 0.0001 compared to the model group.

[0099] #: P < 0.05 compared to the control group; ##: P < 0.01 compared to the control group; ###: P < 0.001 compared to the control group; ####: P < 0.0001 compared to the control group.

[0100] Experimental results: Figure 4 The results showed that, compared with the control group, the gene expression of type I, type III, and type IV collagen was significantly decreased in the model group. Compared with the model group, the concentration of the anti-aging raw material for sensitive skin between 0.125% and 0.5% significantly promoted the gene expression of type I, type III, and type IV collagen, showing a significant difference compared with the model group, and the promoting effect was more significant with increasing concentration.

[0101] The above embodiments are merely examples to illustrate the present invention and are not intended to limit the possible implementations of the invention. Based on the disclosure of this invention, those skilled in the art can make various modifications and adjustments. It is neither possible nor necessary to list all possible implementations. Any modifications, equivalent substitutions, or improvements made within the basic principles and scope of this invention should be considered to fall within the protection scope of this invention.

Claims

1. An anti-aging composition suitable for sensitive skin, characterized in that, The anti-aging composition comprises the following raw materials by weight: 80-95 parts of lactic acid bacteria fermentation extract, 3-4 parts of 1,2-pentanediol, 5-9 parts of 1,3-propanediol, 0.05-0.3 parts of octanoyl hydroxamic acid; The preparation method of the lactic acid bacteria fermentation extract comprises the following steps: S1, crushing dry tricholoma matsutake, adding water at a material-liquid ratio of 1:(10-30), extracting at 80-100℃ for 1-4 hours, and filtering to obtain tricholoma matsutake extract; S2, adding 30-50% ethanol solution to white peony root powder at a material-liquid ratio of 1:(15-40), ultrasonic extraction at 50-80℃ for 1-3 hours, and filtering to obtain white peony root extract; S3, mixing the tricholoma matsutake extract and the white peony root extract at a mass ratio of (3-8):1 to obtain a mixed extract, adding a nitrogen source and inorganic salt to the mixed extract, adjusting the pH to 5.0-7.0, first sterilization, inoculating lactic acid bacteria, and fermenting at 30-42℃ for 24-120 hours, filtering, second sterilization, and obtaining a fermentation liquor; S4, adding a composite enzyme to the fermentation liquor of step S3, ultrasonic reaction, centrifugation, collecting the supernatant, and obtaining the lactic acid bacteria fermentation extract.

2. The anti-aging composition for sensitive skin according to claim 1, wherein The particle size of the tricholoma matsutake powder after crushing in step S1 is 50-100 mesh; the particle size of the white peony root powder in step S2 is 40-80 mesh; and the microwave treatment in step S2 is 300W-600W for 3-10 minutes at 40-60℃.

3. The anti-aging composition for sensitive skin according to claim 1, wherein The inorganic salt in step S3 is one or more of ammonium chloride, potassium dihydrogen phosphate, sodium sulfate, and calcium chloride, and the addition amount is 0.5-3wt% of the mass of the mixed extract; and the nitrogen source is one or more of peptone, yeast extract, or soybean protein, and the addition amount is 3-10wt% of the mass of the mixed extract.

4. The anti-aging composition for sensitive skin according to any one of claims 1 to 3, wherein The lactic acid bacteria in the step S3 are one or more of Lactobacillus plantarum, Lactobacillus paracasei and Lactobacillus acidophilus, and the inoculation amount is 1-10% of the mass of the mixed extract; the concentration of the lactic acid bacteria is 1×10 8 ~1×10 10 CFU / mL.

5. The anti-aging composition for sensitive skin according to any one of claims 1 to 3, wherein The composite enzyme in step S3 is a mixture of papain and cellulase at a mass ratio of 5-10:1-4.

6. The anti-aging composition for sensitive skin according to any one of claims 1 to 3, wherein The filtering in step S3 is performed using a polypropylene microfiltration membrane with a pore size of 0.5-2.5μm; and the first sterilization and the second sterilization are both low-temperature pasteurization at a temperature of 80-100℃ for 30-60 minutes.

7. The anti-aging composition for sensitive skin according to any one of claims 1 to 3, wherein The addition amount of the composite enzyme in step S4 relative to the fermentation liquor is 0.1-0.5wt%, the ultrasonic power is 500-800W, the reaction temperature is 35-45℃, and the reaction time is 1-2 hours.

8. The anti-aging composition for sensitive skin according to any one of claims 1 to 3, wherein The anti-aging composition can further comprise 0.1-1.0 parts of a penetrant, and the penetrant comprises at least one of ceramide, phytosterol, and low-molecular-weight hyaluronic acid.

9. A method for the preparation of an anti-aging composition suitable for sensitive skin according to any one of claims 1-8, characterized in that, Specifically comprising the following steps: stirring the components uniformly, adjusting the pH to 6.5-7.0, and obtaining the anti-aging composition suitable for sensitive skin.

10. A skin care product comprising, The skin care product contains 1-100% of the whitening composition for sensitive skin according to any one of claims 1-8; and the skin care product dosage form is at least one of an aqueous agent, an emulsion, a cream, an essence, or a mask liquid.

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