Anti-oxidation fucoidan composition for delaying skin stem cell aging as well as preparation method and application of anti-oxidation fucoidan composition

Abstract

The invention belongs to the technical field of cosmetics, and particularly discloses an anti-oxidation fucoidan composition for delaying skin stem cell aging and a preparation method and application of the anti-oxidation fucoidan composition. The antioxidant algin composition is prepared from the following components in parts by weight: 5 to 10 parts of algin extract, 5 to 10 parts of herba houttuyniae extract, 2 to 4 parts of black pine needle extract, 4 to 6 parts of sorbitan olivate, 4 to 6 parts of squalane, 5 to 10 parts of glycerol, 0.1 to 0.5 part of sodium hyaluronate, 0.1 to 0.5 part of xanthan gum, 0.1 to 1 part of ethylhexylglycerin and 0.1 to 1 part of p-hydroxyacetophenone. The invention discloses an antioxidant fucoxanthin composition for delaying skin stem cell aging and a preparation method and application thereof. The antioxidant fucoxanthin composition is high in antioxidant capacity and capable of effectively promoting collagen synthesis and delaying skin stem cell aging.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to an antioxidant fucoxanthin composition for delaying skin stem cell aging and a preparation method and application thereof. BACKGROUND

[0002] Skin aging is a complex biological process, one of the core features of which is the imbalance of skin stem cell niche and the decline of stem cell function. Skin stem cells (such as epidermal stem cells and hair follicle stem cells) are responsible for maintaining the self-renewal and damage repair of skin tissue. With age, oxidative stress accumulation, inflammatory microenvironment formation and extracellular matrix (ECM) degradation, skin stem cells undergo aging, showing decreased proliferation, abnormal differentiation and altered secretion profile, ultimately leading to skin atrophy, wrinkle formation, delayed wound healing and other signs of aging. Therefore, targeting skin stem cells, developing active ingredients that can resist oxidative damage and maintain ECM homeostasis has become an important direction of anti-aging research.

[0003] Fucoxanthin is a carotenoid derived from brown algae. Due to its unique propylene bond and epoxy group structure, it has excellent ability to quench singlet oxygen and scavenge free radicals, and is currently recognized as one of the most potent antioxidants. Studies have shown that fucoxanthin can activate the Nrf2 antioxidant pathway and inhibit the expression of matrix metalloproteinases (MMPs), thus showing great potential in delaying skin cell aging. However, fucoxanthin faces two major bottlenecks in practical application: first, its molecular structure is highly unsaturated, and it is extremely sensitive to light, heat and oxygen, easily oxidized and degraded, resulting in a significant loss of bioactivity during formulation processing and storage; second, its strong hydrophobicity results in poor solubility in aqueous media, making it difficult to penetrate the hydrophilic stratum corneum of the skin to reach deep target sites (such as stem cells in the basal layer), severely limiting its bioavailability and skin care efficacy. In addition, relying solely on a single active ingredient often cannot comprehensively intervene in the multi-factor network of aging. How to scientifically compound fucoxanthin with plant extracts to produce a synergistic effect in delaying skin stem cell aging and ensure the stability of the entire composition system remains a technical problem to be solved in the field. SUMMARY

[0004] The present application aims to provide an antioxidant fucoxanthin composition for delaying skin stem cell aging and a preparation method and application thereof. The antioxidant fucoxanthin composition has strong antioxidant capacity, can effectively promote collagen synthesis and delay skin stem cell aging.

[0005] To solve the above technical problems, the technical solution adopted by the present application is as follows: An antioxidant fucoxanthin composition for delaying skin stem cell aging comprises the following components by weight: 5-10 parts of phycocyanin extract, 5-10 parts of houttuynia cordata extract, 2-4 parts of black pine leaf extract, 4-6 parts of sorbitan olive oil ester, 4-6 parts of squalane, 5-10 parts of glycerin, 0.1-0.5 parts of sodium hyaluronate, 0.1-0.5 parts of xanthan gum, 0.1-1 parts of ethylhexylglycerin, and 0.1-1 parts of p-hydroxyacetophenone.

[0006] Preferably, the extraction method of the fucoidan extract includes the following steps: A1. Kelp or kelp is rinsed with pure water, freeze-dried until the moisture content is ≤5%, and then crushed to obtain pre-treated raw materials; A2. Add the pretreated raw material obtained in A1 to the compound enzyme and hydrolyze it to obtain the enzymatic hydrolysate; A3. Pasteurize the enzymatic hydrolysate obtained in A2, cool it to a certain temperature, adjust the pH, inoculate it with a compound strain, and ferment it to obtain the fermentation broth. A4. Centrifuge the fermentation broth obtained in A3, collect the supernatant, and concentrate the supernatant to 1 / 5-1 / 10 of its original volume; freeze-dry the concentrate and pulverize it to obtain the fucoidan extract powder.

[0007] Preferably, in A2, the compound enzyme is a mixture of cellulase, alginate lyase and pectinase in a mass ratio of (2-4):(1-2):1.

[0008] Preferably, in A2, the enzymatic hydrolysis conditions are: temperature of 45-55℃, pH of 5.0-6.5, and time of 2-4h.

[0009] Preferably, in step A3, the enzymatic hydrolysate obtained in step A2 is pasteurized, cooled to 28-37°C, and the pH is adjusted to 6.5-7.0.

[0010] Preferably, in A3, the compound bacterial strain is Bacillus subtilis (… Bacillus subtilis ) and Bacillus licheniformis ( Bacillus licheniformis The total inoculum amount of the mixed strains, composed of a mass ratio of (1-2):1, is 3%-8% of the volume of the enzyme hydrolysate.

[0011] Preferably, the Bacillus subtilis is Bacillus subtilis CCRC 10255, and the effective viable count of the Bacillus subtilis is 1×10⁻⁶. 9 -2×10 9 CFU / g; the Bacillus licheniformis is Bacillus licheniformis ATCC 14580, and the effective viable count of the Bacillus licheniformis is 1×10⁻⁶. 9 -2×10 9 CFU / g.

[0012] Preferably, in A3, the specific conditions of fermentation are as follows: the fermentation temperature is 28-37℃, and the fermentation time is 48-72h.

[0013] Preferably, in A4, the supernatant is concentrated to 1 / 5-1 / 10 of the original volume at 50-60℃ and a vacuum degree of -0.1 MPa.

[0014] Preferably, the extraction method of the Houttuynia cordata extract comprises the following steps: B1, drying Houttuynia cordata to a moisture content of ≤8%, crushing and passing through a 40-60 mesh sieve to obtain Houttuynia cordata coarse powder; B2, adding the Houttuynia cordata coarse powder obtained in B1 to a 30%-50% ethanol solution at a solid-liquid ratio of 1:8-1:15, and ultrasonically extracting at 40-50℃ for 30-60 min at an ultrasonic power of 300-500W, and extracting twice, and then combining the extraction solutions to obtain a crude extract; B3, adjusting the pH of the crude extract obtained in B2 to 5.5-6.5, adding 0.1%-0.5% β-glucosidase, and enzymatically hydrolyzing at 45-50℃ for 2-4h to obtain an enzymatic hydrolysate; B4, filtering the enzymatic hydrolysate obtained in B3, and concentrating the filtrate to 1 / 5-1 / 8 of the original volume at 50-60℃ and a vacuum degree of -0.1 MPa to obtain a concentrated solution; B5, adding 95% ethanol to the concentrated solution obtained in B4 to a final concentration of 70%-80%, and standing at 4-10℃ for 12-24h, and then taking the supernatant, and concentrating and drying to obtain a Houttuynia cordata extract.

[0015] Preferably, the extraction method of the Houttuynia cordata extract comprises the following steps: C1, drying Houttuynia cordata to a moisture content of ≤8%, crushing and passing through a 40-60 mesh sieve to obtain Houttuynia cordata coarse powder; C2, adding the Houttuynia cordata coarse powder obtained in C1 to purified water at a solid-liquid ratio of 1:5-1:15, and sterilizing at 80-90℃ for 30-60 min, and then cooling to obtain a sterilized substrate; C3, adjusting the pH of the sterilized substrate obtained in C2 to 6.0-7.0, and inoculating with a composite strain to obtain a fermentation liquor; C4, filtering the fermentation liquor obtained in C3, taking the liquid part, and concentrating and drying to obtain a Houttuynia cordata extract.

[0016] Preferably, in C3, the composite strain is a mixed strain composed of Bacillus subtilis ( Bacillus subtilis ) and Bacillus licheniformis ( Bacillus licheniformis ) at a mass ratio of (1-2):1, and the total inoculation amount is 3%-8% of the volume of the enzymatic hydrolysate.

[0017] Preferably, the Bacillus subtilis is Bacillus subtilis CCRC 10255, and the effective viable cell number of the Bacillus subtilis is 1×10 9 -2×10 9 CFU / g; the Bacillus licheniformis is Bacillus licheniformis ATCC 14580, and the effective viable cell number of the Bacillus licheniformis is 1×10 9 -2×10 9 CFU / g.

[0018] Preferably, in C3, the specific conditions of fermentation are as follows: the fermentation temperature is 28-37℃, and the fermentation time is 48-72h.

[0019] The application further provides a preparation method of the antioxidant fucoidan composition for delaying skin stem cell aging. S1, dissolving fucoidan extract powder in ethanol with a mass fraction of 70%-75% to obtain a core material solution under light-proof conditions; S2, mixing hydroxypropyl-β-cyclodextrin with deionized water, heating, and stirring uniformly to obtain a cyclodextrin solution; S3, slowly dropping the core material solution obtained in S1 into the cyclodextrin solution in S2 under stirring conditions, and constant-temperature stirring to obtain a mixed solution; S4, placing the mixed solution obtained in S3 in a refrigerator at 4℃ overnight, and performing suction filtration, washing, and low-temperature drying to obtain a water-soluble fucoidan-cyclodextrin inclusion compound; S5, mixing Houttuynia cordata extract and black pine leaf extract, and adding deionized water to stir uniformly to obtain a mixed extract solution; S6, uniformly mixing the mixed extract solution in S5 with the water-soluble fucoidan-cyclodextrin inclusion compound in S4, and then adding sorbitan olivate, squalane, glycerol, sodium hyaluronate, xanthan gum, ethylhexylglycerin and p-hydroxyacetophenone in sequence and stirring uniformly to obtain the antioxidant fucoidan composition.

[0020] Preferably, in S2, the heating temperature is 40-60℃, and the heating time is 10-15min.

[0021] The application further provides application of the antioxidant fucoidan composition for delaying skin stem cell aging in preparation of cosmetics or skin care products.

[0022] Compared with the prior art, the application has the following advantages and technical effects: The application discloses an antioxidant fucoidin composition for delaying skin stem cell aging and a preparation method and application thereof. The fucoidin extract obtained through enzymatic fermentation is compounded with Houttuynia cordata and black pine leaf extract, and remarkable synergistic effects are exhibited in terms of antioxidation and promotion of collagen synthesis. Experimental results show that the hydroxyl radical removal rate of the composition is as high as 78.2%, and the expression amount of type IV collagen in fibroblasts damaged by siRNA is significantly increased from 32.4 ng / mL to 68.5 ng / mL, and the repair effect is close to that of the positive control vitamin C. Moreover, the composition adopts a mild preservative system, has high safety, and has no skin irritation and allergic reactions.

[0023] The technical solutions of the application are further described in detail below with the aid of drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 Hydroxyl radical removal rates of antioxidant fucoidin compositions provided for Examples 1-3 and Comparative Examples 1-3; Figure 2 Statistical diagrams of type IV collagen contents of antioxidant fucoidin compositions provided for Examples 1-3 and Comparative Examples 1-3. DETAILED DESCRIPTION

[0025] The technical solutions of the application are further described in detail below with the aid of drawings and examples.

[0026] Unless otherwise defined, the technical terms or scientific terms used in the application shall have the usual meanings understood by those skilled in the art to which the application belongs.

[0027] Sources of test materials: In the application, unless otherwise specified, other test materials and instruments and equipment are conventional test materials in the art, and can be purchased through commercial channels.

[0028] Example 1 An antioxidant fucoidin composition for delaying skin stem cell aging: The antioxidant fucoidin composition comprises the following components in parts by weight: 7 parts of fucoidin extract, 7 parts of Houttuynia cordata extract, 3 parts of black pine leaf extract, 5 parts of sorbitan olivate, 5 parts of squalane, 8 parts of glycerol, 0.3 parts of sodium hyaluronate, 0.3 parts of xanthan gum, 0.5 parts of ethylhexylglycerin, and 0.5 parts of p-hydroxyacetophenone.

[0029] The extraction method of the fucoidin extract comprises the following steps: A1, the kelp is washed with pure water, freeze-dried at -20 DEG C to a water content of less than or equal to 5%, crushed through a 40-mesh sieve, and obtained as pretreated raw material; A2, the pretreated raw material obtained in A1 is added with a complex enzyme agent (cellulase: alginate lyase: pectinase = 3: 1.5: 1, mass ratio), pH is adjusted to 6.0, and enzymolysis is carried out at 50℃ for 3h to obtain an enzymolysis liquid; A3, the enzymolysis liquid obtained in A2 is subjected to pasteurization, cooled to 32℃, pH is adjusted to 6.8, inoculated with a complex strain (Bacillus subtilis CCRC 10255 and Bacillus licheniformis ATCC 14580, mass ratio 1.5: 1, total inoculation amount 5%), and fermented at 32℃ for 60h to obtain a fermentation liquid, wherein the effective viable cell count of Bacillus subtilis and Bacillus licheniformis is 1×10 9 CFU / g; A4, the fermentation liquid obtained in A3 is centrifuged (4000rpm, 15min), and the supernatant is collected and concentrated to 1 / 8 of the original volume under the condition of 55℃ and vacuum degree-0.1MPa; the concentrated liquid is freeze-dried (-50℃, 24h) and pulverized to obtain a laminarin extract powder.

[0030] The extraction method of the extract of Houttuynia cordata Thunb. comprises the following steps: B1, Houttuynia cordata Thunb. is air-dried to a water content of ≤8%, crushed and passed through a 40-mesh sieve to obtain Houttuynia cordata Thunb. coarse powder; B2, the Houttuynia cordata Thunb. coarse powder is taken, 40% ethanol solution is added at a solid-liquid ratio of 1:12, and ultrasonic extraction is carried out at 45℃ for 45min (ultrasonic power 400W), and the extraction is carried out twice, and the extract is obtained by combining the extractives; B3, the extract obtained in B2 is adjusted to pH 6.0, and 0.3% β-glucosidase is added, and enzymolysis is carried out at 48℃ for 3h to obtain an enzymolysis liquid; B4, the enzymolysis liquid obtained in B3 is filtered, and the filtrate is concentrated to 1 / 6 of the original volume under the condition of 55℃ and vacuum degree-0.1MPa to obtain a concentrated liquid; B5, 95% ethanol is added to the concentrated liquid obtained in B4 to a final concentration of 75%, and the mixture is placed at 4℃ for 18h, and the supernatant is taken and concentrated and dried under reduced pressure at 50℃ to obtain a Houttuynia cordata Thunb. extract.

[0031] The extraction method of the extract of Pinus thunbergii Franch. leaves comprises the following steps: C1, Pinus thunbergii Franch. leaves are air-dried to a water content of ≤8%, crushed and passed through a 40-mesh sieve to obtain Pinus thunbergii Franch. leaf coarse powder; C2, the Pinus thunbergii Franch. leaf coarse powder is taken, and pure water is added at a solid-liquid ratio of 1:10, and sterilized at 85℃ for 45min, and then cooled to room temperature to obtain a sterilized substrate; C3, adjust the pH of the sterilized substrate obtained in C2 to 6.5. Inoculate the complex bacterial strain (Bacillus subtilis CCRC 10255 and Bacillus licheniformis ATCC 14580, mass ratio 1.5:1, total inoculation amount 5%). Fermentation conditions: temperature 32℃, time 60h, obtain fermentation broth, wherein the effective viable count of Bacillus subtilis and Bacillus licheniformis is 1×10 9 CFU / g; C4, filter the fermentation broth obtained in C3, take the liquid part, concentrate under reduced pressure at 55℃, then freeze-dry (-50℃, 24h) to obtain black pine leaf extract.

[0032] The antioxidant fucoidan composition for delaying skin stem cell aging, the preparation method comprising the following steps S1, under light-proof conditions, take 7 parts of fucoidan extract powder, dissolve with 72% ethanol by mass fraction to obtain a core material solution; S2, take 15 parts of hydroxypropyl-β-cyclodextrin and mix with 60 parts of deionized water, heat to 50℃, stir uniformly (300rpm, 12min) to obtain a cyclodextrin solution; S3, under stirring conditions (500rpm), slowly drop the core material solution obtained in S1 into the cyclodextrin solution of S2, stir at a constant temperature of 50℃ for 2h to obtain a mixed solution; S4, place the mixed solution obtained in S3 in a refrigerator at 4℃ overnight, suction filter, wash with a small amount of anhydrous ethanol, and dry at a low temperature of 40℃ for 5h to obtain a water-soluble fucoidan-cyclodextrin inclusion complex.

[0033] S5, mix 7 parts of Houttuynia cordata extract and 3 parts of black pine leaf extract, add 20 parts of deionized water, and stir uniformly to obtain a mixed extract solution; S6, mix the mixed extract solution of S5 with the water-soluble fucoidan-cyclodextrin inclusion complex of S4, then sequentially add sorbitan olivate 5 parts, squalane 5 parts, glycerol 8 parts, sodium hyaluronate 0.3 parts, xanthan gum 0.3 parts, ethylhexylglycerin 0.5 parts, and p-hydroxyacetophenone 0.5 parts, and stir uniformly to obtain an antioxidant fucoidan composition.

[0034] Example 2 The preparation method is the same as that of Example 1, except that the antioxidant fucoidan composition comprises the following components by weight: fucoidan extract 10 parts, Houttuynia cordata extract 10 parts, black pine leaf extract 4 parts, sorbitan olivate 6 parts, squalane 6 parts, glycerol 10 parts, sodium hyaluronate 0.5 parts, xanthan gum 0.5 parts, ethylhexylglycerin 1 part, and p-hydroxyacetophenone 1 part.

[0035] Example 3 The preparation method is the same as that in Example 1, except that the antioxidant fucoidin composition comprises the following components by weight: fucoidin extract 5 parts, Houttuynia cordata extract 5 parts, black pine leaf extract 2 parts, sorbitan olivate 4 parts, squalane 4 parts, glycerol 5 parts, sodium hyaluronate 0.1 part, xanthan gum 0.1 part, ethylhexylglycerin 0.1 part, and p-hydroxyacetophenone 0.1 part.

[0036] Comparative Example 1 An antioxidant fucoidin composition for delaying skin stem cell aging, comprising the following components by weight: fucoidin extract 7 parts, Houttuynia cordata extract 7 parts, sorbitan olivate 5 parts, squalane 5 parts, glycerol 8 parts, sodium hyaluronate 0.3 part, xanthan gum 0.3 part, ethylhexylglycerin 0.5 part, and p-hydroxyacetophenone 0.5 part.

[0037] An extraction method of fucoidin extract, comprising the following steps: A1, kelp is washed with pure water, freeze-dried at -20℃ to a water content of ≤5%, crushed through a 40-mesh sieve, and obtained as pretreated raw material; A2, the pretreated raw material obtained in A1 is added with a composite enzyme agent (cellulase:fucoidin lyase:pectinase=3:1.5:1, mass ratio), the pH is adjusted to 6.0, and enzyme hydrolysis is performed at 50℃ for 3h to obtain an enzyme hydrolysate; A3, the enzyme hydrolysate obtained in A2 is subjected to pasteurization, cooled to 32℃, the pH is adjusted to 6.8, inoculated with a composite bacterial strain (Bacillus subtilis CCRC 10255 and Bacillus licheniformis ATCC 14580 at a mass ratio of 1.5:1, total inoculation amount of 5%), and fermented at 32℃ for 60h to obtain a fermentation broth, wherein the effective viable bacterial count of Bacillus subtilis and Bacillus licheniformis is 1×10 9 CFU / g; A4, the fermentation broth obtained in A3 is centrifuged (4000rpm, 15min), the supernatant is collected, and concentrated to 1 / 8 of the original volume under the conditions of 55℃ and vacuum degree of -0.1MPa; the concentrated solution is freeze-dried (-50℃, 24h), and crushed to obtain fucoidin extract powder.

[0038] An extraction method of Houttuynia cordata extract, comprising the following steps: B1, Houttuynia cordata is air-dried to a water content of ≤8%, crushed through a 40-mesh sieve, and obtained as Houttuynia cordata coarse powder; B2, the Houttuynia cordata coarse powder is taken, 40% ethanol solution is added at a solid-liquid ratio of 1:12, ultrasonic extraction is performed at 45℃ for 45min (ultrasonic power 400W), and the extraction is performed twice, and the extract solutions are combined to obtain a crude extract solution; B3, the crude extract solution obtained in B2 is adjusted to pH 6.0, 0.3% β-glucosidase is added, and enzyme hydrolysis is performed at 48℃ for 3h to obtain an enzyme hydrolysate; B4, filter the enzymatic hydrolysate obtained in B3, and concentrate the filtrate to 1 / 6 of the original volume at 55℃ under a vacuum of-0.1 MPa to obtain a concentrated solution; B5, add 95% ethanol to the concentrated solution obtained in B4 to a final concentration of 75%, stand at 4℃ for 18 h, take the supernatant, and concentrate and dry the supernatant at 50℃ under reduced pressure to obtain the extract of Houttuynia cordata.

[0039] An antioxidant fucoidan composition for delaying skin stem cell aging, and a preparation method thereof, comprising the following steps S1, under light-proof conditions, dissolve 7 parts of fucoidan extract powder in 72% ethanol by mass fraction to obtain a core material solution; S2, mix 15 parts of hydroxypropyl-β-cyclodextrin with 60 parts of deionized water, heat to 50℃, and stir uniformly (300 rpm, 12 min) to obtain a cyclodextrin solution; S3, under stirring conditions (500 rpm), slowly drop the core material solution obtained in S1 into the cyclodextrin solution of S2, and stir at a constant temperature of 50℃ for 2 h to obtain a mixed solution; S4, store the mixed solution obtained in S3 in a refrigerator at 4℃ overnight, filter, wash with a small amount of anhydrous ethanol, and dry at a low temperature of 40℃ for 5 h to obtain a water-soluble fucoidan-cyclodextrin inclusion compound.

[0040] S5, mix 7 parts of the extract of Houttuynia cordata with 20 parts of deionized water to obtain a mixed extract solution; S6, mix the mixed extract solution of S5 with the water-soluble fucoidan-cyclodextrin inclusion compound of S4, and then sequentially add sorbitan olivate 5 parts, squalane 5 parts, glycerol 8 parts, sodium hyaluronate 0.3 parts, xanthan gum 0.3 parts, ethylhexylglycerin 0.5 parts, and p-hydroxyacetophenone 0.5 parts, and stir uniformly to obtain an antioxidant fucoidan composition.

[0041] Comparative Example 2 The same as in Example 1, except that: A method for extracting fucoidan extract, comprising the following steps: A1, rinse kelp with purified water, freeze-dry at-20℃ to a water content of ≤5%, and crush through a 40-mesh sieve to obtain pretreated raw material; A2, add a composite enzyme agent (cellulase:fucoidan lyase:pectinase=3:1.5:1 by mass ratio) to the pretreated raw material obtained in A1, adjust the pH to 6.0, and enzymatically hydrolyze at 50℃ for 3 h to obtain an enzymatic hydrolysate; A3, centrifuging the enzymatic hydrolysate obtained in A2 (4000 rpm, 15 min), collecting the supernatant, concentrating to 1 / 8 of the original volume at 55°C under a vacuum of -0.1 MPa, freeze-drying the concentrated solution (-50°C, 24 h), and pulverizing, to obtain the laminarin extract powder.

[0042] Comparative Example 3 The same as in Example 1, except that: The extraction method of the laminarin extract comprises the following steps: A1, washing the kelp with purified water, freeze-drying at -20°C until the water content is ≤5%, pulverizing through a 40-mesh sieve, and obtaining the pretreated raw material; A2, adding a composite enzyme agent (cellulase: alginate lyase: pectinase = 3: 1.5: 1, mass ratio) to the pretreated raw material obtained in A1, adjusting the pH to 6.0, and enzymatically hydrolyzing at 50°C for 3 h, to obtain an enzymatic hydrolysate; A3, centrifuging the enzymatic hydrolysate obtained in A2 (4000 rpm, 15 min), collecting the supernatant, concentrating to 1 / 8 of the original volume at 55°C under a vacuum of -0.1 MPa, freeze-drying the concentrated solution (-50°C, 24 h), and pulverizing, to obtain the laminarin extract powder.

[0043] The antioxidant laminarin compositions provided in Examples 1-3 and Comparative Examples 1-3 above were subjected to effect verification.

[0044] Antioxidant test: The antioxidant laminarin compositions provided in Examples 1-3 and Comparative Examples 1-3 were added to deionized water, respectively, to prepare sample solutions at 0.5 mg / mL. The specific test groups are as follows: Experimental group: The antioxidant laminarin compositions prepared in Examples 1, 2, and 3 were set as groups A, B, and C, respectively.

[0045] Control group: The compositions prepared in Comparative Examples 1, 2, and 3 were set as groups D, E, and F, respectively.

[0046] Positive control: Vitamin C (Vc) was set as group G.

[0047] Blank control: Deionized water was set as group H.

[0048] Then 400 μL of 9 mmoL / L ferrous sulfate solution, 400 μL of sample solution, and 400 μL of 9 mmoL / L H2O2 solution were sequentially added to a 2 mL centrifuge tube, mixed, and placed at room temperature for 15 min, 300 μL of 9 mmoL / L ethanol-salicylic acid solution was added, shaken, and placed in a 37°C water bath for 25 min, the absorbance value was measured at 510 nm, each experiment was repeated 3 times, and the average value was taken.

[0049] The hydroxyl radical scavenging rate is calculated according to the following formula: ; Wherein: : absorbance of the sample solution; : absorbance measured by using deionized water instead of the H2O2 solution; : absorbance of the sample solution measured by using deionized water instead of the H2O2 solution.

[0050] The results are shown in Table 1. Figure 1

[0051] As can be seen from Table 1, the antioxidant fucoidan composition provided by the embodiments 1-3 has good antioxidant performance. Figure 1

[0052] The promoting effect of the composition on the synthesis of extracellular matrix (ECM) by skin fibroblasts is evaluated by detecting the expression amount of type IV collagen.

[0053] Experimental cells: HFF-1 (human skin fibroblasts).

[0054] The HFF-1 cells in the logarithmic growth phase were taken, and the cell density was adjusted to 2×10 5 The cells were inoculated in a 24-well plate, 500 μL of cell suspension was added to each well, and the plate was placed in a 37℃, 5% CO2 incubator for culture for 24 h, so that the cells were adherent and fused to about 70%. According to the transfection reagent instruction, the siRNA working solution (final concentration 50 nM) targeting human COL4A1 gene was prepared (H group, blank control group without siRNA, and the same amount of culture medium was added). The original culture medium was aspirated, and 200 μL of siRNA working solution was added to each well for 6 h of incubation with the cells. The siRNA working solution was aspirated, and 500 μL of the sample solution prepared in advance was added to each well. The sample solution was prepared into 0.1 mg / mL with deionized water, filtered through a 0.22 μm filter membrane, and sterilized. The same volume of serum-free medium was added to the H group and the model control group. The culture was continued for 36 h, and the cell culture supernatant of each well was collected. The operation was strictly performed according to the instruction of the human type IV collagen ELISA kit, and the content of type IV collagen (ng / mL) in the supernatant of each well was determined.

[0055] The specific test groups are as follows: Experimental group: the antioxidant fucoidan compositions prepared in examples 1, 2 and 3, which are respectively set as groups A, B and C.

[0056] Control group: the compositions prepared in comparative examples 1, 2 and 3, which are respectively set as groups D, E and F.

[0057] ​​Positive control: Vitamin C (Vc), designated as group G.

[0058] Blank control: Deionized water, set as group H.

[0059] Model group: siRNA + serum-free culture medium, set as group M.

[0060] The results are as follows Figure 2 As shown.

[0061] Depend on Figure 2 It can be seen that the antioxidant fucoidin compositions obtained in Examples 1-3 of the present invention have a good effect on promoting the secretion of type IV collagen in cells.

[0062] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. An antioxidant fucoidan composition for delaying the aging of skin stem cells, characterized in that, Includes the following components by weight: 5-10 parts of phycocyanin extract, 5-10 parts of houttuynia cordata extract, 2-4 parts of black pine leaf extract, 4-6 parts of sorbitan olive oil ester, 4-6 parts of squalane, 5-10 parts of glycerin, 0.1-0.5 parts of sodium hyaluronate, 0.1-0.5 parts of xanthan gum, 0.1-1 parts of ethylhexylglycerin, and 0.1-1 parts of p-hydroxyacetophenone.

2. The antioxidant fucoidan composition according to claim 1, characterized in that, The extraction method for the fucoidan extract includes the following steps: A1. Kelp or kelp is rinsed with pure water, freeze-dried until the moisture content is ≤5%, and then crushed to obtain pre-treated raw materials; A2. Add the pretreated raw material obtained in A1 to the compound enzyme and hydrolyze it to obtain the enzymatic hydrolysate; A3. Pasteurize the enzymatic hydrolysate obtained in A2, cool it to a certain temperature, adjust the pH, inoculate it with a compound strain, and ferment it to obtain the fermentation broth. A4. Centrifuge the fermentation broth obtained in A3, collect the supernatant, and concentrate the supernatant to 1 / 5-1 / 10 of its original volume; freeze-dry the concentrate and pulverize it to obtain the fucoidan extract powder.

3. The antioxidant fucoidan composition according to claim 2, characterized in that, In A2, the compound enzyme is a mixture of cellulase, alginate lyase and pectinase in a mass ratio of (2-4):(1-2):

1.

4. The antioxidant fucoidan composition according to claim 2, characterized in that, In A2, the specific enzymatic hydrolysis conditions are: temperature 45-55℃, pH 5.0-6.5, and time 2-4h.

5. The antioxidant fucoidan composition according to claim 2, characterized in that, In A3, the compound strain is a mixture of Bacillus subtilis and Bacillus licheniformis in a mass ratio of (1-2):1, and the total inoculum amount is 3%-8% of the volume of the enzymatic hydrolysate.

6. The antioxidant fucoidan composition according to claim 5, characterized in that, The Bacillus subtilis strain is Bacillus subtilis CCRC 10255, and the effective viable count of the Bacillus subtilis strain is 1×10⁻⁶. 9 -2×10 9 CFU / g; the Bacillus licheniformis is Bacillus licheniformis ATCC 14580, and the effective viable count of the Bacillus licheniformis is 1×10⁻⁶. 9 -2×10 9 CFU / g.

7. The antioxidant fucoidan composition according to claim 2, characterized in that, In A3, the specific fermentation conditions are: a fermentation temperature of 28-37℃ and a fermentation time of 48-72h.

8. A method for preparing an antioxidant fucoidin composition for delaying skin stem cell aging as described in claim 1, characterized in that, Includes the following steps: S1. Under light-protected conditions, dissolve the fucoidan extract powder in 70%-75% ethanol to obtain a core material solution. S2. Mix hydroxypropyl-β-cyclodextrin with deionized water, heat, and stir until homogeneous to obtain a cyclodextrin solution; S3. Under stirring conditions, the core material solution obtained in S1 is slowly added dropwise to the cyclodextrin solution in S2, and stirred at a constant temperature to obtain a mixture. S4. The mixture obtained in S3 was placed in a refrigerator at 4°C overnight, filtered, washed, and dried at low temperature to obtain a water-soluble fucoidan-cyclodextrin inclusion complex. S5. Mix the houttuynia cordata extract and black pine leaf extract, add deionized water, stir well to obtain a mixed extract; S6. Mix the mixed extract of S5 with the water-soluble fucoidan-cyclodextrin inclusion complex of S4 until homogeneous, then add sorbitan olive oil ester, squalane, glycerol, sodium hyaluronate, xanthan gum, ethylhexylglycerin and p-hydroxyacetophenone in sequence, and stir until homogeneous to obtain the antioxidant fucoidan composition.

9. The preparation method according to claim 8, characterized in that, In S2, the heating temperature is 40-60℃ and the heating time is 10-15 minutes.

10. The use of the antioxidant fucoidin composition for delaying skin stem cell aging as described in claim 1 in the preparation of cosmetics or skin care products.

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