A high-content radix pseudostellariae cyclic peptide b radix pseudostellariae extract, a preparation method thereof and a cosmetic use thereof

Abstract

The application discloses a high-content Pseudostellaria heterophylla cyclic peptide B Pseudostellaria heterophylla extract and a preparation method and cosmetic use thereof, and belongs to the technical field of cosmetics. The preparation method comprises the following steps: (1) crushing Pseudostellaria heterophylla and adding alcohol to perform ultrasonic treatment to obtain a feed liquid; (2) adding a composite enzyme into the feed liquid to perform enzymolysis, enzyme inactivation, solid-liquid separation, concentration and impurity removal to obtain an enzymolysis liquid; and (3) performing macroporous resin elution on the enzymolysis liquid to obtain an eluate, and drying the eluate to obtain the Pseudostellaria heterophylla cyclic peptide B Pseudostellaria heterophylla extract. The composite enzyme comprises pectinase, papain, beta-glucosidase, cellulase and xylanase, and the mass ratio of the pectinase, the papain, the beta-glucosidase, the cellulase and the xylanase is 5-10:0.5-1:1:3-7:0.3-1. The Pseudostellaria heterophylla extract is prepared by using an enzymolysis process, more active ingredients can be reserved, the content of Pseudostellaria heterophylla cyclic peptide B in the obtained extract is high, and the obtained extract has good anti-inflammatory, soothing, repairing and anti-aging effects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a Codonopsis pilosula extract with a high content of Codonopsis pilosula cyclic peptide B, its preparation method, and its cosmetic uses. Background Technology

[0002] Prince ginseng is a member of the Caryophyllaceae family. Pseudostellaria heterophylla The dried tuberous root of (Miq.) Pax ex PaxHoffm.; harvested in summer when the stems and leaves wither, washed, fibrous roots removed, briefly blanched in boiling water, and then sun-dried or directly sun-dried. Codonopsis pilosula has traditional effects such as invigorating qi and strengthening the spleen, promoting body fluids and moistening the lungs; it is mainly used to treat symptoms such as spleen deficiency and fatigue, weakness after illness, qi and yin deficiency, spontaneous sweating and thirst, and dry cough due to lung dryness.

[0003] Modern research shows that Codonopsis pilosula mainly contains chemical components such as cyclic peptides, saponins, sugars, amino acids, phospholipids, volatile oils, fatty acids, oils, sterols, and trace elements. It has the effects of myocardial protection, immune regulation, antioxidation, hypoglycemia, anti-stress, anti-fatigue, anti-tumor, and antitussive. It is widely used in health food, pharmaceuticals, and daily chemical products.

[0004] Currently, most extraction methods for *Pseudostellaria heterophylla* involve water extraction, which results in significant loss of active ingredients and affects its efficacy to some extent. CN109528803A discloses a method for preparing *Pseudostellaria heterophylla* extract, including water extraction followed by freeze-drying of the resulting extract. This invention found that freeze-drying after water extraction can reduce the loss of active ingredients in the extract to some extent, but the content of cyclic peptides is low, especially the content of *Pseudostellaria heterophylla* cyclic peptide B, which is only 0.0002-0.0003%.

[0005] Xiong Hejian et al. (Study on in vitro antioxidant activity of Codonopsis pilosula extract [J]. Journal of Nankai University (Natural Science Edition), December 2009, 42(6), 37-41) disclosed that the water extract and alcohol extract of Codonopsis pilosula have significant antioxidant activity, and the alcohol extract is significantly better than the water extract, but they did not disclose the effect of different extraction processes on the content of Codonopsis pilosula cyclic peptide B. CN107929127A discloses a method for preparing a Codonopsis pilosula extract, comprising the following steps: 1) Preparation of water-soluble substances from Codonopsis pilosula: Weigh dried Codonopsis pilosula medicinal material, pulverize it, soak it in water overnight, reflux for extraction, filter, and obtain filtrate; 2) Preparation of non-polar substances from Codonopsis pilosula: Use a low-melting-point solvent betaine-glycerol mixed solution as the extraction solvent to perform ultrasonic-assisted extraction of non-polar substances from the residue after water extraction of Codonopsis pilosula, filter, and obtain filtrate B; 3) Combination of water-soluble and non-polar substances from Codonopsis pilosula: Combine filtrate A and filtrate B, and concentrate under reduced pressure on a rotary evaporator to obtain the Codonopsis pilosula extract. Although this invention has a good extraction effect on cyclic peptides, it does not disclose the effect on the content of Codonopsis pilosula cyclic peptide B. SUMMARY

[0006] The present application aims to provide a high-content Pseudostellaria heterophylla cyclic peptide B Pseudostellaria heterophylla extract and its preparation method and cosmetic use, which is prepared by enzymatic hydrolysis, can retain more active ingredients, has high content of Pseudostellaria heterophylla cyclic peptide B in the prepared Pseudostellaria heterophylla extract, has good anti-inflammatory, anti-aging, soothing and repairing effects, and can be used as an effective raw material in cosmetics.

[0007] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0008] In the first aspect, the present application provides a Pseudostellaria heterophylla extract prepared by enzymatic hydrolysis of Pseudostellaria heterophylla as a substrate with a composite enzyme.

[0009] In some embodiments, the composite enzyme is pectinase, papain, beta-glucosidase, cellulase and xylanase; preferably, the mass ratio of the pectinase, papain, beta-glucosidase, cellulase and xylanase is 5-10:0.5-1:1:3-7:0.3-1; further preferably 7-9:0.5-0.7:1:4-6:0.4-0.6; more preferably 8-9:0.6-0.7:1:4-5:0.5-0.6; most preferably 8:0.6:1:5:0.5.

[0010] In the second aspect, the present application provides a preparation method of the above-mentioned Pseudostellaria heterophylla extract, comprising the following steps:

[0011] (1) Pseudostellaria heterophylla is crushed, alcohol ultrasonic treatment is performed, alcohol is recovered, and a feed liquid is obtained;

[0012] (2) A composite enzyme is added to the feed liquid for enzymatic hydrolysis, enzyme is inactivated, solid-liquid separation is performed, and impurities are removed by concentration to obtain an enzymatic hydrolysis liquid;

[0013] (3) The enzymatic hydrolysis liquid is eluted by a macroporous resin to obtain an eluate, and the eluate is dried to obtain the Pseudostellaria heterophylla extract.

[0014] In some embodiments, the alcohol is added in step (1) in an amount of 8-10 times the mass of Pseudostellaria heterophylla, preferably 9 times; the temperature of the alcohol is 40-50°C, preferably 45-50°C; the alcohol is an ethanol aqueous solution with a volume fraction of 30-70%, preferably a volume fraction of 45-70%.

[0015] In some embodiments, the ultrasonic treatment in step (1) is ultrasonic treatment at 300-500W for 12-25min, preferably 400W for 20min.

[0016] In some embodiments, the complex enzyme in step (2) is pectinase, papain, beta-glucosidase, cellulase and xylanase; preferably, the mass ratio of the pectinase, papain, beta-glucosidase, cellulase and xylanase is 5-10:0.5-1:1:3-7:0.3-1; further preferably 7-9:0.5-0.7:1:4-6:0.4-0.6; more further preferably 8-9:0.6-0.7:1:4-5:0.5-0.6; most further preferably 8:0.6:1:5:0.5.

[0017] In some embodiments, the amount of the complex enzyme added in step (2) is 0.1%-0.3% of the weight of the alcohol; preferably 0.2%.

[0018] In some embodiments, the enzymolysis in step (2) is carried out at 40-50°C for 2-4h; preferably at 45°C±2°C for 3h.

[0019] In some embodiments, the solid-liquid separation in step (2) is to separate the solid phase and the liquid phase, and thus, conventional technical means such as centrifugation, filtration or pressure filtration can be used.

[0020] In some embodiments, the concentration and impurity removal in step (2) is to concentrate to a relative density of 1.10-1.12 at 60°C, store at -2-4°C for 18-30h, filter, and obtain the enzymolysis solution.

[0021] In some embodiments, the macroporous resin in step (3) is AB-8, D101 or HPD100. Preferably, it is AB-8.

[0022] In some embodiments, the elution in step (3) is: the sample liquid volume is 1-5BV, the sample is loaded at a flow rate of 0.5-4BV / h, and the elution is carried out with 2-8BV of 50-80% ethanol at a flow rate of 1-5BV / h. Preferably, the sample liquid volume is 2.5BV, the sample is loaded at a flow rate of 1BV / h, and the elution is carried out with 3BV of 70% ethanol at a flow rate of 2BV / h.

[0023] In some embodiments, the drying in step (3) is carried out at a temperature not higher than 50°C, preferably freeze drying, reduced pressure drying or spray drying.

[0024] In some embodiments, the preparation method of the Radix Pseudostellariae extract comprises the following steps:

[0025] (1) The Radix Pseudostellariae is crushed, 8-10 times the amount of alcohol (30-70% ethanol aqueous solution) is added, and ultrasonic treatment is carried out at 300-500W for 12-25min to obtain a slurry;

[0026] (2) adding a complex enzyme to the feed liquid, and carrying out enzymolysis at 40-50°C for 2-4h, inactivating the enzyme after the reaction, and then carrying out solid-liquid separation, concentrating the liquid phase to a relative density of 1.10-1.12 at 60°C, and then storing at -2-4°C for 18-30h, and then filtering to remove impurities, to obtain an enzyme hydrolysate;

[0027] The complex enzyme is added in an amount of 0.1%-0.3% of the weight of the alcohol; and the complex enzyme comprises pectinase, papain, β-glucosidase, cellulase and xylanase in a mass ratio of 5-10:0.5-1:1:3-7:0.3-1.

[0028] (3) subjecting the enzyme hydrolysate to macroporous resin elution, subjecting the sample liquid to elution at a flow rate of 0.5-4BV / h, and then eluting with 2-8BV of 50%-80% ethanol at a flow rate of 1-5BV / h, to obtain an eluate, and then drying the eluate.

[0029] In a third aspect, the present application provides a use of a complex enzyme in increasing the content of Taizishen cyclic peptide B in a Taizishen extract.

[0030] In some embodiments, the complex enzyme comprises pectinase, papain, β-glucosidase, cellulase and xylanase; preferably, the mass ratio of the pectinase, papain, β-glucosidase, cellulase and xylanase is 5-10:0.5-1:1:3-7:0.3-1; further preferably, the mass ratio is 7-9:0.5-0.7:1:4-6:0.4-0.6; more preferably, the mass ratio is 8-9:0.6-0.7:1:4-5:0.5-0.6; most preferably, the mass ratio is 8:0.6:1:5:0.5.

[0031] In a fourth aspect, the present application provides a use of the above-mentioned Taizishen extract in preparing a cosmetic product.

[0032] The Taizishen extract of the present application has a high content of Taizishen cyclic peptide B, and can be used as a functional raw material in a cosmetic product. For example, the functional raw material has at least one of the effects of anti-inflammation, soothing, repair and anti-aging.

[0033] In a fifth aspect, the present application further provides a cosmetic product comprising the above-mentioned Taizishen extract.

[0034] In some embodiments, the cosmetic product is in a form of milk, cream, water, powder, oil or block.

[0035] The cosmetic product described in the present application can contain, in addition to the above-mentioned functional raw materials (Radix Pseudostellariae extract), one or more optional ingredients known or otherwise effective for use in personal care products, provided that these optional ingredients are compatible with the physical and chemical properties of the essential ingredients described herein, or do not excessively affect the stability, aesthetics or performance of the product. Non-limiting examples of such optional ingredients are described in the "Cosmetic Safety Technical Specifications, 2015 Edition", which is incorporated herein by reference.

[0036] The present application has the following beneficial effects:

[0037] (1) The present application uses pectinase, papain, beta-glucosidase, cellulase and xylanase to jointly enzymatically hydrolyze Radix Pseudostellariae water extract, so that more active ingredients can be retained. Compared with the prior art, the content of Pseudostellariae cyclic peptide B in the Radix Pseudostellariae extract prepared by the present application is higher, which can reach 0.13%-0.33%.

[0038] (2) The experiment proves that the Radix Pseudostellariae extract prepared by the present application has good safety, and has good anti-inflammatory, soothing, repairing and anti-aging effects.

[0039] (3) The preparation process of the present application is simple, the conditions are mild, the cost is low, and it is suitable for large-scale popularization and application. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 It is a representative diagram of the tail neutral cells of zebrafish embryos, and the dark dots are the stained neutrophils;

[0041] Figure 2 It is a schematic diagram of the measurement area of skin barrier damage of zebrafish embryos, and the skin barrier damage is marked with green fluorescence;

[0042] Figure 3 It is a test result diagram of skin barrier permeability of zebrafish embryos. DETAILED DESCRIPTION

[0043] The following description of the embodiments is merely exemplary in nature and is in no way intended to limit the scope of the application, its application, or its core teachings. It should be noted that for one of ordinary skill in the art, with the benefit of this disclosure, numerous modifications and alterations to the described embodiments are possible without departing from the teachings of the present application. Numerous modifications and alterations to the described embodiments will readily occur to those skilled in the art. Accordingly, the disclosure is intended to embrace all such alterations and modifications as fall within the scope of the present application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0044] The present application does not limit the source of raw materials used, such as no special instructions, the raw materials used in the present application are ordinary commercially available in the art. The used radix pseudostellariae in the present application is produced in Guizhou, preferably Huangping and Shibing.

[0045] Unless otherwise specified, all operations herein are carried out at room temperature (20-25°C) environment, the solvent used is water, the amount of the present application is relative to the weight of the raw material, for example, 9 times the amount of alcohol in example 1, the specific amount of alcohol is 9 times the mass of radix pseudostellariae.

[0046] Below, the examples and comparative examples of the present application are shown, and the radix pseudostellariae extract of the present application is described in more detail, but the present application is not limited thereto, unless otherwise specified, the percentage in the present application is mass percentage. Pectinase, item YB30778; papain, item YB49326; β-glucosidase, item YB124915; cellulase, item YB37524; xylanase, item YB125042, are all purchased from Hunan Yunbang Biotechnology Co., Ltd.

[0047] Example 1 Preparation method of radix pseudostellariae extract

[0048] The steps are as follows:

[0049] (1) Radix pseudostellariae is crushed through a 60-mesh sieve, 9 times the amount of alcohol (60% ethanol aqueous solution by volume, temperature 45°C) is added, and ultrasonic extraction is carried out at 400W for 20min. The ethanol is recovered to obtain the feed liquid;

[0050] (2) adding the compound enzyme into the liquid, and carrying out enzymolysis at 45°C for 3h; after the reaction, the enzyme is inactivated by keeping at 121°C for 15min; centrifugation is carried out at 1500rpm for 10min to obtain a centrifugal liquid; the centrifugal liquid is concentrated to have a relative density of 1.10-1.12 at 60°C; the concentrated liquid is stored at 0°C for 24h; and the enzyme hydrolysis liquid is obtained by passing through a 0.22μm acetic acid membrane;

[0051] The adding amount of the compound enzyme is 0.2% of the weight of the alcohol; the compound enzyme is pectinase, papain, β-glucosidase, cellulase and xylanase with a mass ratio of 8:0.6:1:5:0.5;

[0052] (3) the enzyme hydrolysis liquid is eluted by AB-8 macroporous resin; the volume of the sample liquid is 2.5BV; the sample is loaded at a flow rate of 1BV / h; 3BV of 70% ethanol is used for elution at a flow rate of 2BV / h; the eluted liquid is obtained; and the eluted liquid is dried under reduced pressure (50°C, 0.05Mpa) to obtain the product.

[0053] Preparation method of the extract of radix pseudostellariae in example 2

[0054] The steps are as follows:

[0055] (1) the radix pseudostellariae is crushed to pass through a 60-mesh sieve; 8 times of alcohol (30% ethanol aqueous solution, 40°C) is added; ultrasonic treatment is carried out at 300W for 25min; and the alcohol is recovered to obtain a liquid;

[0056] (2) the compound enzyme is added into the liquid; and enzymolysis is carried out at 40°C for 4h; after the reaction, the enzyme is inactivated by keeping at 121°C for 15min; centrifugation is carried out at 1500rpm for 10min to obtain a centrifugal liquid; the centrifugal liquid is concentrated to have a relative density of 1.10-1.12 at 60°C; the concentrated liquid is stored at 0°C for 24h; and the enzyme hydrolysis liquid is obtained by passing through a 0.22μm acetic acid membrane;

[0057] The adding amount of the compound enzyme is 0.2% of the weight of the alcohol; the compound enzyme is pectinase, papain, β-glucosidase, cellulase and xylanase with a mass ratio of 8:0.6:1:5:0.5;

[0058] (3) the enzyme hydrolysis liquid is eluted by AB-8 macroporous resin; the volume of the sample liquid is 2.5BV; the sample is loaded at a flow rate of 1BV / h; 3BV of 70% ethanol is used for elution at a flow rate of 2BV / h; the eluted liquid is obtained; and the eluted liquid is dried under reduced pressure (50°C, 0.05Mpa) to obtain the product.

[0059] Preparation method of the extract of radix pseudostellariae in example 3

[0060] The steps are as follows:

[0061] (1) The Prince Shenjiang is crushed through a 60-mesh sieve, 10 times the amount of alcohol (70% ethanol aqueous solution by volume, temperature 50°C) is added, and ultrasonic is performed at 500W for 12min. The ethanol is recovered to obtain a liquid;

[0062] (2) The compound enzyme is added to the liquid, and enzymolysis is performed at 50°C for 2h. After the reaction is completed, the enzyme is inactivated by keeping at 121°C for 15min, and centrifugation is performed at 1500rpm for 10min to obtain a centrifugal liquid. The centrifugal liquid is concentrated to a relative density of 1.10-1.12 at 60°C, and is stored at 0°C for 24h. The 0.22μm acetic acid membrane is used to obtain an enzymolysis liquid;

[0063] The addition amount of the compound enzyme is 0.3% of the weight of the alcohol; the compound enzyme is pectinase, papain, β-glucosidase, cellulase and xylanase with a mass ratio of 10:1:1:7:1;

[0064] (3) The enzymolysis liquid is eluted by AB-8 macroporous resin, the sample liquid volume is 5BV, the flow rate is 4BV / h, 8BV of 80% ethanol is used for elution at a flow rate of 5BV / h, and an elution liquid is obtained. The elution liquid is dried under reduced pressure (50°C, 0.05Mpa) to obtain the product.

[0065] Preparation method of Prince Shenjiang extract

[0066] The difference between this example and Example 1 is that the mass ratio of pectinase, papain, β-glucosidase, cellulase and xylanase is different.

[0067] The specific steps are as follows:

[0068] (1) The Prince Shenjiang is crushed through a 60-mesh sieve, 10 times the amount of alcohol (70% ethanol aqueous solution by volume, temperature 50°C) is added, and ultrasonic is performed at 500W for 12min. The ethanol is recovered to obtain a liquid;

[0069] (2) The compound enzyme is added to the liquid, and enzymolysis is performed at 50°C for 2h. After the reaction is completed, the enzyme is inactivated by keeping at 121°C for 15min, and centrifugation is performed at 1500rpm for 10min to obtain a centrifugal liquid. The centrifugal liquid is concentrated to a relative density of 1.10-1.12 at 60°C, and is stored at 0°C for 24h. The 0.22μm acetic acid membrane is used to obtain an enzymolysis liquid;

[0070] The addition amount of the compound enzyme is 0.3% of the weight of the alcohol; the compound enzyme is pectinase, papain, β-glucosidase, cellulase and xylanase with a mass ratio of 10:1:1:7:1;

[0071] (3) The enzyme solution is eluted by AB-8 macroporous resin, the sample solution volume is 2.5BV, the sample is loaded at a flow rate of 1BV / h, and 3BV of 70% ethanol is used for elution at a flow rate of 2BV / h, to obtain an eluate, and the eluate is dried under reduced pressure (50°C, 0.05Mpa) to obtain the product.

[0072] Preparation method of Prince seng's extract in Comparative Example 2

[0073] The difference between this comparative example and Example 1 is that the mass ratio of pectinase, papain, beta-glucosidase, cellulase and xylanase is different.

[0074] The specific steps are as follows:

[0075] (1) The Prince seng is crushed to pass through a 60-mesh sieve, 9 times the amount of alcohol (60% ethanol aqueous solution by volume, temperature 45°C) is added, and ultrasonic extraction is performed at 400W for 20min, and the ethanol is recovered to obtain a feed solution;

[0076] (2) The compound enzyme is added to the feed solution, and enzymolysis is performed at 45°C for 3h, and after the reaction is completed, the enzyme is inactivated by keeping at 121°C for 15min, and centrifugation is performed at 1500rpm for 10min to obtain a centrifugal solution, which is concentrated to a relative density of 1.10-1.12 at 60°C, and then stored at 0°C for 24h, and then filtered through a 0.22μm acetic acid membrane to obtain an enzyme solution;

[0077] The addition amount of the compound enzyme is 0.2% of the weight of the alcohol; the compound enzyme is pectinase, papain, beta-glucosidase, cellulase and xylanase with a mass ratio of 15:3:0.8:0.5:0.2;

[0078] (3) The enzyme solution is eluted by AB-8 macroporous resin, the sample solution volume is 2.5BV, the sample is loaded at a flow rate of 1BV / h, and 3BV of 70% ethanol is used for elution at a flow rate of 2BV / h, to obtain an eluate, and the eluate is dried under reduced pressure (50°C, 0.05Mpa) to obtain the product.

[0079] Preparation method of Prince seng's extract in Comparative Example 3

[0080] The difference between this comparative example and Example 1 is that there is no enzyme hydrolysis step.

[0081] The specific steps are as follows:

[0082] (1) The Prince seng is crushed to pass through a 60-mesh sieve, 9 times the amount of alcohol (60% ethanol aqueous solution by volume, temperature 45°C) is added, and ultrasonic extraction is performed at 400W for 20min, and the ethanol is recovered to obtain a feed solution;

[0083] (2) centrifugal liquid 1500 rpm under centrifugal 10 min, get centrifugal liquid, centrifugal liquid concentrated to 60 ℃ under the relative density of 1.10-1.12, 0 ℃ under refrigeration 24 h, 0.22 μm acetic acid membrane, get clear liquid;

[0084] (3) clear liquid elution by AB-8 macroporous resin, sample liquid volume is 2.5BV, with 1BV / h flow rate, elution with 3BV, 70% ethanol with 2BV / h flow rate, get elution liquid, elution liquid reduced pressure drying (50 ℃, 0.05Mpa), get.

[0085] I. Content of Taizishen cyclic peptide B in Taizishen extract

[0086] The Taizishen extract provided by the examples and comparative examples is detected by high performance liquid chromatography (HPLC) for the content of Taizishen cyclic peptide B according to the content determination of 2010 edition of Chinese Pharmacopoeia, and the results are shown in Table 1.

[0087] Table 1

[0088]

[0089] The results show that the content of Taizishen cyclic peptide B in the Taizishen extract prepared by the examples 1-3 of the present application reaches 0.13%-0.33%.

[0090] Meanwhile, comparing the single factor experiments of comparative example 1, comparative example 2 and example 1, when the mass ratio of pectinase, papain, β-glucosidase, cellulase and xylanase is not in the range claimed in the present application, the content of Taizishen cyclic peptide B in the prepared Taizishen extract is only 0.0018%-0.0020%, which shows that the specific mass ratio of pectinase, papain, β-glucosidase, cellulase and xylanase in the present application can significantly improve the content of Taizishen cyclic peptide B in the Taizishen extract.

[0091] II. L929 cell activity test

[0092] The blank group (DMEM medium), cell control group (DMEM medium+cells) and sample group (DMEM medium+cells+Taizishen extract prepared by examples 1-3 and comparative examples 1-3) are set for cell activity test, and each group has 3 parallel holes.

[0093] The logarithmic growth period of mouse fibroblast L929 cells is 1×10 5The cells were seeded in 96-well plates at a density of 1 x 104 / mL. Incubation was carried out at 37°C for 24 hours. The culture solution was removed, 100 μL of sample solution of different concentrations was added to each well, and incubation was continued at 37°C for 12 hours. After 20 μL of 5 mg / mL MTT solution was added to each well and mixed, incubation was continued at 37°C for 2 hours. The supernatant was discarded, and 100 μL of isopropanol was added to each well. Oscillation was carried out at a speed of 100 r / min for 30 minutes in the dark at room temperature. The absorbance of each well was detected at a wavelength of 570 nm using an enzyme-labeled instrument.

[0094] The cell viability was calculated according to the following formula,

[0095] ;

[0096] In the formula, OD (sample group) represents the absorbance of the sample group, OD (blank group) represents the absorbance of the blank group, and OD (cell control group) represents the absorbance of the cell control group.

[0097] The results are shown in Tables 2 and 3.

[0098] Table 2

[0099]

[0100] The results show that, compared with the blank group, the cell viability is significantly increased at concentrations of 2.50 mg / mL, 5.00 mg / mL and 100 mg / mL (P value < 0.001 or < 0.0001). This indicates that the extract of Radix Pseudostellariae prepared in Example 1 has a cell proliferation effect at the above-mentioned concentrations.

[0101] Table 3

[0102]

[0103] The results show that, compared with the blank group, the cell viability of the Example 1-Example 3 groups is significantly increased (P value < 0.001); while the cell viability of the Comparative Example 1-Comparative Example 3 groups has no significant change (P value > 0.05), indicating that the extract of Radix Pseudostellariae prepared in Example 1-Example 3 has a cell proliferation effect at a concentration of 5.00 mg / mL, while the extract of Radix Pseudostellariae prepared in Comparative Example 1-Comparative Example 3 has no cell proliferation effect.

[0104] III. Expression of SIRT1 Longevity Protein in L929 Cells

[0105] The experiment was set up with a blank group (DMEM medium), a model group (DMEM medium + cells + H2O2) and a sample group (DMEM medium + cells + H2O2 + extract of Radix Pseudostellariae prepared in Example 1-Example 3 and Comparative Example 1-Comparative Example 3), with 3 parallel holes in each group.

[0106] Mouse fibroblast L929 cells in logarithmic growth phase were inoculated in 96-well plates at a density of 1 x 10 5 Individual wells were added with 100 μL of sample solution at different concentrations, and incubated at 37 °C for 12 hours. SIRT1 protein expression was tested using a SIRT1 Elisa kit.

[0107] The results are shown in Table 4.

[0108] Table 4

[0109]

[0110] The results show that the SIRT1 protein expression of the cells in the Example 1-Example 3 groups was significantly increased (P value < 0.0001 or 0.01) compared with the model group; while the SIRT1 protein expression of the cells in the Comparative Example 1-Comparative Example 3 groups had no significant change (P value > 0.05), indicating that the extract of P. notoginseng prepared in Example 1-Example 3 had the effect of enhancing the expression of SIRT1 protein of the cells at a concentration of 5.00 mg / mL.

[0111] Four, L929 cell phospho-STAT3D protein expression

[0112] The experiment was set up with a blank group (DMEM medium), a model group (DMEM medium + cells + H2O2), and a sample group (DMEM medium + cells + H2O2 + the extract of P. notoginseng prepared in Example 1-Example 3 and Comparative Example 1-Comparative Example 3), with 3 parallel holes in each group.

[0113] Mouse fibroblast L929 cells in logarithmic growth phase were inoculated in 96-well plates at a density of 1 x 10 5 Individual wells were added with 100 μL of sample solution at different concentrations, and incubated at 37 °C for 12 hours. SIRT1 protein expression was tested using a SIRT1 Elisa kit.

[0114] The results are shown in Table 5.

[0115] Table 5

[0116]

[0117] The results show that: compared with the model group, the expression of P-STAT3 protein in the cells of the example 1 to example 3 groups is significantly reduced (P value < 0.0001 or 0.05); while the expression of P-STAT3 protein in the cells of the comparative example 1 to comparative example 3 groups has no significant change (P value > 0.05), indicating that the extract of radix pseudostellariae prepared in the example 1 to example 3 has an inhibitory effect on the expression of P-STAT3 protein in the cells at the concentration of 5.00 mg / mL.

[0118] V. Nematode Anti-lipid Aging and Anti-oxidative Stress

[0119] Cell test model: Caenorhabditis elegans.

[0120] Nematode Anti-lipid Aging Test Method: After synchronization of the nematodes, the L4 stage nematodes were intervened with 20 mg / mL of the samples for 10 days, and 20 mg / mL of vitamin C was used as a positive control, each group was in triplicate, and after sodium azide anesthesia, the nematodes were transferred to a 2% agarose gel pad, and the fluorescence pictures were obtained under a fluorescence microscope, and the fluorescence intensity was analyzed.

[0121] The results are shown in Table 6.

[0122] Table 6

[0123]

[0124] The results show that: compared with the blank group, the relative fluorescence intensity of lipofuscin in the nematodes of the example 1 to example 3 groups is significantly reduced (P value < 0.0001 or 0.01); while the relative fluorescence intensity of lipofuscin in the nematodes of the comparative example 1 to comparative example 3 groups has no significant change (P value > 0.05), indicating that the extract of radix pseudostellariae prepared in the example 1 to example 3 has an inhibitory effect on the expression of lipofuscin in the nematodes at the concentration of 20 mg / mL.

[0125] Nematode Anti-oxidative Stress Test Method: After synchronization of the nematodes, the L4 stage nematodes were transferred to the above blank group, positive control group or sample group for 5 days, and then transferred to a new NGM plate, and the nematodes were exposed to the plate containing hydrogen peroxide. The survival rate under stress was calculated.

[0126] The results are shown in Table 7.

[0127] Table 7

[0128]

[0129] The results show that: compared with the blank group, the average lifespan of nematodes in the example 1 to example 3 groups is significantly increased (P value < 0.05); while the average lifespan of nematodes in the positive control group, the comparative example 1 to comparative example 3 groups has no significant change (P value > 0.05), indicating that the extract of radix pseudostellariae prepared in example 1 to example 3 has the effect of prolonging the lifespan of nematodes under oxidative stress at a concentration of 20 mg / mL.

[0130] VI. Zebrafish Anti-aging Test

[0131] 6.1 Zebrafish Sirt1 Gene Expression Promotion Test

[0132] Sirt1 is an NAD+ (nicotinamide adenine dinucleotide) dependent deacetylase, which is involved in various cellular biological functions mainly through deacetylation of various non-histone and histone proteins, and is a recognized longevity gene. Studies have found that when mammalian and zebrafish Sirt1 genes are knocked out or defective, reactive oxygen species ROS increases, chronic inflammation, and lifespan shortens; and promoting Sirt1 gene expression can improve metabolism and overall health of the organism. Therefore, the anti-aging efficacy of a sample can be evaluated by testing whether the sample promotes Sirt1 gene expression.

[0133] In this experiment, a blank control group (fish embryo culture solution), a positive control group (glutathione) and a test substance group (the extract of radix pseudostellariae provided in example 1) were set. Specifically:

[0134] Blank control group: 36 zebrafish were randomly selected and evenly distributed in a 24-well plate, each well containing 12 zebrafish and 2.5 mL of fish embryo culture solution;

[0135] Positive control group: 36 zebrafish were randomly selected and evenly distributed in a 24-well plate, 3 wells were set, each well containing 12 zebrafish and 2.5 mL of glutathione working solution (0.08 mg / mL glutathione solution prepared with fish embryo culture solution).

[0136] Test substance group: 36 zebrafish were randomly selected and evenly distributed in a 24-well plate, 3 wells were set, each well containing 12 zebrafish and 2.5 mL of test substance solution (0.2% extract of radix pseudostellariae in example 1 solution prepared with fish embryo culture solution).

[0137] Placed in a constant temperature incubator at 28℃±1℃ and cultured to 24h±1h after fertilization. 12 zebrafish in each well were collected in a 1.5 mL tube, the solution was removed, and 0.5 mL of RNAlater solution (Invitrogen; AM7020) was added for ice storage.

[0138] RNA was extracted from zebrafish, and RNA was synthesized into cDNA using PrimeScript RT kit (Takara; Cat no. RR047A) containing gDNA Eraser, and real-time PCR amplification was performed using SYBR Premix Ex Taq kit (Takara; Cat no. RR420A), with β-actin as the housekeeping gene, Ct as the amplification result, and the relative expression of Sirt1 was calculated and statistically analyzed, using 2 -△△Ct The gene expression promotion rate of Sirt1 was calculated according to the following formula.

[0139] ;

[0140] In the formula, 2 -△△Ct 受试物 : the relative expression of the gene of interest of the test group; 2 -△△Ct 空白对照组 : the relative expression of the gene of interest of the blank control group.

[0141] The results are shown in Table 8.

[0142] Table 8

[0143]

[0144] The results show that the promotion rate of the extract of Pseudostellaria heterophylla prepared in Example 1 on the expression of Sirt1 gene of zebrafish is 75% at an addition concentration of 0.2%, indicating that the extract of Pseudostellaria heterophylla prepared in the application can promote the expression of Sirt1 gene of zebrafish and has an anti-aging effect.

[0145] 6.2, Zebrafish elastin gene (Elna) expression promotion test

[0146] Elastin in human skin is a protein that allows many tissues in the body to maintain their original shape after stretching and contraction, and allows the skin to return to its original shape after being squeezed. With age, elastin fibers in the skin gradually break down and are lost, causing wrinkles and loss of elasticity in the skin. Compared with human skin, zebrafish skin is highly similar to human skin in structure, except that the epidermis is composed of differentiated epidermal cells and a mucous layer rather than a stratum corneum. Elastin is highly conserved, and elastin in the human body is synthesized by the Eln gene, and the corresponding gene of the Elna gene in zebrafish. By testing whether the sample promotes the expression of the Elna gene in zebrafish, it can be determined whether the sample promotes the regeneration of elastin, helps the skin to resist wrinkles, and achieves the effect of skin tightening. Many domestic and foreign cosmetics achieve the effects of skin anti-wrinkle and tightening by promoting the regeneration of elastin.

[0147] The experiment sets blank control group (fish embryo culture solution), positive control group (acetyl hexapeptide-8) and test substance group (the extract of radix pseudostellariae provided in Example 1). Specifically:

[0148] Blank control group: 36 zebrafish were randomly selected and evenly distributed in 24-well plates, each well containing 12 zebrafish and 2.5 mL of fish embryo culture solution;

[0149] Positive control group: 36 zebrafish were randomly selected and evenly distributed in 24-well plates, 3 wells were set up, each well containing 12 zebrafish and 2.5 mL of acetyl hexapeptide-8 working solution (0.8 g / L acetyl hexapeptide-8 solution prepared with fish embryo culture solution).

[0150] Test substance group: 36 zebrafish were randomly selected and evenly distributed in 24-well plates, 3 wells were set up, each well containing 12 zebrafish and 2.5 mL of test substance solution (0.2% radix pseudostellariae extract solution prepared with fish embryo culture solution).

[0151] Place in a constant temperature incubator at 28℃±1℃ and cultivate until 24h±1h after fertilization. Collect 12 zebrafish in each well in a 1.5 mL tube, remove the solution, and add 0.5 mL of RNAlater solution (Invitrogen; AM7020) and store in the freezer.

[0152] RNA extraction was performed on zebrafish, RNA was synthesized into cDNA using PrimeScript RT Kit with gDNA Eraser (Takara; Cat no. RR047A), real-time PCR amplification was performed using SYBR Premix Ex Taq Kit (Takara; Cat no. RR420A), β-actin was used as the housekeeping gene, Ct was used as the amplification result, the relative expression of elastin gene (Elna) was calculated and statistically analyzed, and 2 -△△Ct The gene expression promotion rate of Elna was calculated according to the following formula.

[0153] ;

[0154] In the formula, 2 -△△Ct 受试物 : relative expression of target gene in test substance group; 2 -△△Ct 空白对照组 : relative expression of target gene in blank control group.

[0155] The results are shown in Table 9.

[0156] Table 9

[0157]

[0158] The results show that the promotion rate of the extract of Radix Pseudostellariae prepared in Example 1 on the expression of the Elna gene of zebrafish is 20% at an addition concentration of 0.2%, indicating that the extract of Radix Pseudostellariae prepared in the application can promote the expression of the Elna gene of zebrafish and has the effect of promoting the regeneration of elastin, so as to achieve the effects of anti-wrinkle and firming.

[0159] 6.3, Promotion test of zebrafish type I collagen gene expression

[0160] Collagen is the highest content of extracellular matrix protein in human body, and type I collagen is the most abundant protein in the skin. The distribution of type I collagen of zebrafish is the same as that of human body, and shows high conservation with human. Their innate expression significantly decreases at 6 days after fertilization and becomes very low between 10 to 12 days after fertilization. The expression of zebrafish type I collagen genes (col1a1a, col1a1b and col1a2) is tested, the relative expression amount of zebrafish type I collagen genes of the treatment group and the blank control group is compared, and the promotion rate of type I collagen gene expression is calculated to evaluate the anti-wrinkle and firming effects of raw materials or products.

[0161] In this experiment, a blank control group (fish embryo culture solution), a positive control group (acetyl hexapeptide-8) and a test substance group (the extract of Radix Pseudostellariae provided in Example 1) are set. Specifically:

[0162] Blank control group: 36 zebrafish are randomly selected and evenly distributed in a 24-well plate, each well containing 12 zebrafish and 2.5 mL of fish embryo culture solution;

[0163] Positive control group: 36 zebrafish are randomly selected and evenly distributed in a 24-well plate, 3 wells are set, and each well contains 12 zebrafish and 2.5 mL of acetyl hexapeptide-8 working solution (0.8 g / L acetyl hexapeptide-8 solution prepared with fish embryo culture solution).

[0164] Test substance group: 36 zebrafish are randomly selected and evenly distributed in a 24-well plate, 3 wells are set, and each well contains 12 zebrafish and 2.5 mL of test substance solution (0.2% extract of Radix Pseudostellariae solution prepared with fish embryo culture solution).

[0165] Place in a constant temperature incubator at 28℃±1℃ and cultivate to 24h±1h after fertilization. Collect 12 zebrafish in each well in a 1.5 mL tube, remove the solution, and add 0.5 mL of RNAlater solution (Invitrogen; AM7020) for ice storage.

[0166] RNA was extracted from zebrafish, and RNA was synthesized into cDNA using PrimeScript RT kit (Takara; Cat no. RR047A) containing gDNA Eraser, and real-time PCR amplification was performed using SYBR Premix Ex Taq kit (Takara; Cat no. RR420A), with β-actin as the housekeeping gene, Ct as the amplification result, the relative expression of collagen type I gene was calculated and statistically analyzed, and 2 -△△Ct According to the following formula, the gene expression promotion rate of collagen type I gene was calculated.

[0167] ;

[0168] In the formula, 2 -△△Ct 受试物 : relative expression of the gene of interest of the test group; 2 -△△Ct 空白对照组 : relative expression of the gene of interest of the blank control group.

[0169] The results are shown in Table 10.

[0170] Table 10

[0171]

[0172] Note: compared with the blank control group, **P<0.01.

[0173] The results show that the promotion rates of the extract of Pseudostellaria heterophylla prepared in Example 1 on the expression of zebrafish col1a1a, col1a1b and col1a2 genes are 62%, 56% and 56% respectively at an addition concentration of 0.2%, indicating that the extract of Pseudostellaria heterophylla prepared in the application can promote the expression of zebrafish collagen type I gene, has the effect of promoting collagen type I protein regeneration, and can realize the effects of anti-wrinkle and firming.

[0174] Seven, zebrafish embryo neutrophil test

[0175] Soothing refers to helping to improve the state of skin irritation, etc. When the skin is stimulated by physical, chemical or bacteria invasion, it will produce an immune response, appear red and swollen, etc. and show signs of sensitive skin. At this time, immune white blood cells (including neutrophils and macrophages) appear in the skin to phagocytose and remove harmful substances such as infection, foreign matter or damaged cells in the skin. Inhibition of white blood cell aggregation at the stimulated site can soothe the skin irritation state. Zebrafish embryo neutrophils (the main immune white blood cells) and human neutrophils are highly similar in morphology, biochemistry and physiological function, therefore, the test results obtained by establishing a stimulated zebrafish model have good extrapolation to human skin efficacy. Zebrafish embryo neutrophils can be visualized in vivo by fluorescently labeled neutrophil transgenic lines or observed by Sudan black staining. In this test, a model of copper sulfate-induced stimulation of neural crest cells in the lateral line region of zebrafish embryos to cause neutrophil aggregation was used for testing. The number of neutrophils in the lateral line region of the test material group and the model control group of fish embryos was compared, and the neutrophil aggregation inhibition rate was calculated to evaluate whether the raw material, formula or product has soothing effect to soothe irritation and achieve soothing effect.

[0176] The experiment set up a blank control group, a model control group, a positive control group and a test material group (the extract of radix pseudostellariae provided in Example 1). Specifically:

[0177] Blank control group: 24 zebrafish embryos were randomly selected into a 3 cm culture dish, and 5 mL of fish embryo culture solution was added;

[0178] Model control group: 24 zebrafish embryos were randomly selected into a 3 cm culture dish, and 5 mL of fish embryo culture solution containing 10 μM anhydrous copper sulfate was added;

[0179] Positive control group: 24 zebrafish embryos were randomly selected into a 3 cm culture dish, and 5 mL of fish embryo culture solution containing 10 μM anhydrous copper sulfate and 10 μM indomethacin was added;

[0180] Test material group: 24 zebrafish embryos were randomly selected into a 3 cm culture dish, and 5 mL of fish embryo culture solution containing 10 μM anhydrous copper sulfate and 0.2% test material was added.

[0181] Place in a 28℃±1℃ incubator for 40-45min. Fix the fish embryos in paraformaldehyde for at least 1h, then treat the fish embryos with PBST for 3 times, 5min each time, and then treat the fish embryos with 50% ethanol for 3min. After staining the fish embryos with Sudan black staining solution at room temperature for 1h, immerse the fish embryos in 70% ethanol for 4 times, 5min each time, and then treat the fish embryos with PBST for 2 times, 5min each time. Treat the fish embryos with bleaching solution for 10min, and keep the lid open if the treatment is performed in a test tube. Then treat the fish embryos with 70% ethanol solution for 5min, PBST for 1min, clear solution 1 for 15min, clear solution 2 for 10min, and PBST for 3min. Place the fish embryos on their sides. Then place the fish embryos under a stereomicroscope to take pictures of the tail of the fish embryos, and count the number of neutrophils in the lateral line area of the fish embryos from the anus to one-third of the tail (see Figure 1

[0182] Calculate the inhibition rate of neutrophil aggregation as follows:

[0183]

[0184] wherein S is the average number of neutrophils in the test group fish embryos, in units of particles per tail (particles / tail); M is the average number of neutrophils in the model control group fish embryos, in units of particles per tail (particles / tail); and B is the average number of neutrophils in the blank control group fish embryos, in units of particles per tail (particles / tail).

[0185] The results are shown in Table 11.

[0186] Table 11

[0187]

[0188] Note: compared with the model control group, ***P<0.01.

[0189] The results show that the inhibition rate of neutrophil aggregation in the model control group zebrafish embryos is 0, and the zebrafish embryos do not have soothing effect. The inhibition rate of neutrophil aggregation in the zebrafish embryos treated with 10μM indomethacin is 39%, and the inhibition rate of neutrophil aggregation in the zebrafish embryos treated with the extract of radix pseudostellariae prepared in Example 1 at a concentration of 0.2% is 92%. This shows that the extract of radix pseudostellariae prepared in Example 1 of the present application can significantly inhibit the aggregation of neutrophils in zebrafish embryos, and has soothing effect.

[0190] Eight, zebrafish embryo skin barrier permeability test

[0191] ​​24 zebrafish embryos at 72 h were exposed to 0.2% formula added concentration of radix pseudostellariae extract solution, while setting up blank control group and model control group, after 1 h of exposure, fish embryos were co-treated with sodium dodecyl sulfate solution (SDS) + sample solution for 40 min, then stained with sodium fluorescein for 30 min, fluorescence was photographed to measure the tail fin fluorescence intensity and statistical analysis was performed.

[0192] Specifically, the experiment set up blank control group, model control group, positive control group and test substance group (radix pseudostellariae extract provided in Example 1). Among them,

[0193] Blank control group: 24 zebrafish embryos were randomly selected into a 6-well plate, and 6 mL of fish embryo culture solution was added;

[0194] Model control group: 24 zebrafish embryos were randomly selected into a 6-well plate, and 6 mL of fish embryo culture solution was added;

[0195] Positive control group: 24 zebrafish embryos were randomly selected into a 6-well plate, and 6 mL of ikkduin working solution (150 mg / L, prepared with fish embryo culture solution) was added;

[0196] Test substance group: 24 zebrafish embryos were randomly selected into a 6-well plate, and 6 mL of test substance solution (containing 0.2% radix pseudostellariae extract provided in Example 1 fish embryo culture solution) was added.

[0197] After placing in 28℃±1℃ for 1h±0.1h, the blank control group was replaced with 6 mL of new fish embryo culture solution, and the rest of the groups were replaced with 6 mL of fish embryo culture solution containing 0.05 g / L SDS, and placed in 28℃±1℃ for 40 min. After washing the fish embryos 3 times, they were stained with 1 g / L sodium fluorescein solution (prepared with fish embryo culture solution) for 30 min.

[0198] Open the photos with analysis software, mark the tail fin area (see Figure 2 ) of each fish embryo, and select the "average signal intensity" in the test results as the fluorescence intensity.

[0199] Calculate the skin barrier permeability inhibition rate:

[0200] ;

[0201] In the formula, S is the average value of the tail fin fluorescence intensity of the fish embryos in the test substance treatment group; M is the average value of the tail fin fluorescence intensity of the fish embryos in the model control group; B is the average value of the tail fin fluorescence intensity of the fish embryos in the blank control group.

[0202] The results are shown in Figure 3 and Table 12.

[0203] Table 12

[0204]

[0205] Note: compared with the model control group, ***P<0.01.

[0206] The results show that the skin barrier permeability inhibition rate of the model control group is 0, and it has no repair effect; the skin barrier permeability inhibition rate of 150 mg / L icodextrin is 41%; the skin barrier permeability inhibition rate of the extract of Pseudostellaria heterophylla prepared in Example 1 at a concentration of 0.2% is 30%, which indicates that the extract of Pseudostellaria heterophylla prepared in Example 1 can significantly reduce the skin permeability of zebrafish embryos and has the effect of repairing the skin barrier, and can support the repair efficacy.

[0207] The above is a further description of the present application in combination with specific embodiments, but these embodiments are only exemplary and do not constitute any limitation on the scope of the present application. Those skilled in the art should understand that the details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and such modifications and replacements all fall within the protection scope of the present application.

Claims

1. A method for preparing a high-content Codonopsis lanceolata cyclic peptide B extract of Codonopsis lanceolata, characterized by, It comprises the following steps: (1) crushing the radix pseudostellariae, adding alcohol and performing ultrasonic treatment, recovering the alcohol, and obtaining a feed liquid; (2) adding a compound enzyme to the feed liquid for enzymolysis, inactivating the enzyme, performing solid-liquid separation, concentrating and removing impurities, and obtaining an enzymolysis liquid; (3) eluting the enzymolysis liquid through a macroporous resin to obtain an eluate, and drying to obtain the radix pseudostellariae extract; In step (1), the alcohol is an alcohol aqueous solution with a volume fraction of 30%-70%; In step (2), the compound enzyme is a mixture of pectinase, papain, β-glucosidase, cellulase and xylanase with a mass ratio of 5-10:0.5-1:1:3-7:0.3-1; In step (3), the macroporous resin is AB-8; In step (3), the elution is performed by using 50%-80% ethanol.

2. The production method according to claim 1, characterized by, In step (1), the alcohol is added in an amount of 8-10 times the mass of the radix pseudostellariae, and the temperature of the alcohol is 40-50°C; In step (1), the ultrasonic treatment is performed at 300-500W for 12-25min.

3. The method of claim 1, wherein, In step (2), the compound enzyme is added in an amount of 0.1%-0.3% of the weight of the alcohol; In step (2), the enzymolysis is performed at 40-50°C for 2-4h; In step (2), the concentration and impurity removal are performed by concentrating to a relative density of 1.10-1.12 at 60°C, cold storage at -2-4°C for 18-30h, and filtration to remove impurities to obtain the enzymolysis liquid.

4. The method of claim 1, wherein, The mass ratio of the pectinase, papain, β-glucosidase, cellulase and xylanase is 7-9:0.5-0.7:1:4-6:0.4-0.

6.

5. The preparation method according to claim 1, characterized in that, In step (3), the elution is performed by loading a liquid volume of 1-5BV, loading at a flow rate of 0.5-4BV / h, eluting with 2-8BV of 50%-80% ethanol at a flow rate of 1-5BV / h.

6. The radix pseudostellariae extract prepared by the preparation method of any one of claims 1-5.

7. The use of complex enzyme combination in improving the content of Radix Pseudostellariae cyclic peptide B in Pseudostellaria heterophylla extract, characterized in that, The compound enzyme is pectinase, papain, β-glucosidase, cellulase and xylanase, and the mass ratio of the pectinase, papain, β-glucosidase, cellulase and xylanase is 5-10:0.5-1:1:3-7:0.3-1.

8. Use of the radix pseudostellariae extract prepared by the preparation method of any one of claims 1-5 in the preparation of a cosmetic.

9. Use according to claim 8, characterized in that, The cosmetic is in the form of milk, cream, water, powder, oil or block.

10. A cosmetic product, characterized by, The cosmetic comprises the radix pseudostellariae extract prepared by the preparation method of any one of claims 1-5.

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