Atractylodes rhizome root polysaccharide and application thereof

Abstract

The application belongs to the technical field of plant polysaccharides, and discloses a rhizoma atractylodis root polysaccharide and application. The specific structure of the polysaccharide with a molecular weight of 3.7 kD in the rhizoma atractylodis root is determined, the corresponding functional properties are studied, and the application of the polysaccharide with the specific structure in skin anti-aging activity is provided. The anti-aging activity of a product can be improved by promoting the proliferation of keratinocyte HaCAT, inhibiting the beta-galactosidase activity in the keratinocyte HaCAT and ROS active oxygen generation.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant polysaccharides, and particularly relates to a rhizoma atractylodis rootlet polysaccharide and application thereof. BACKGROUND

[0002] The main cause of skin aging is the combined effect of imbalance of the body's antioxidant system, collagen damage, long-term excessive exposure to sunlight, etc., which causes the generation of phenomena such as affecting cell activity, reducing collagen and elastin synthesis, and increasing collagen degradation. The aging skin presents fine wrinkles, dryness, thinning, and increased temperature sensitivity. With the increasing improvement of people's living standards, consumers have higher pursuit of anti-aging products. Skin protection is carried out through various measures such as internal administration of "health products" and external use of "skincare products", and especially, many consumers accept cosmetic skincare products. At present, anti-aging skincare products in the skincare product market are mainly face creams, which are added with ingredients such as A alcohol, peptides, recombinant collagen and bovine growth factors, and have strong irritation, poor stability, slow effect, high price and certain preparation technical requirements, so the development of anti-aging skincare products still has great challenges.

[0003] Rhizoma atractylodis (Atractylodes chinensis (DC.) Koidz.) is a plant of the genus Atractylodes of the family Asteraceae, and the rhizome thereof mainly contains polyphenols, flavonoids and polysaccharides, etc., and has anti-inflammatory, antioxidant, antibacterial, anti-tumor and other biological activities, and has extremely high medicinal value. In recent years, the application of rhizoma atractylodis polysaccharides has been reported more, but the application in antioxidant and anti-aging has not been reported. Compared with the rhizome, the associated rootlet is a non-drug part, and is often discarded in the production process. The rhizome atractylodes rootlet is homologous to the rhizome, and may also contain similar active ingredients. It is found through preliminary research that the polysaccharide content in the rhizome atractylodes rootlet is high, and the rhizome atractylodes rootlet polysaccharide has the advantages of low toxicity, good water solubility and easy development. It is reported that small molecular weight plant polysaccharides have the characteristics of good permeability, high bioavailability and small side effects, are more easily penetrate the skin barrier, play an anti-aging role in the deep layer of the skin, and are more easily absorbed and utilized by skin cells, and can be used as a safe and effective anti-aging ingredient suitable for people of various skin types. At the same time, the structure and anti-aging related activity of the small molecular polysaccharide component in the rhizome atractylodes rootlet have not been reported. The present application aims to determine the structure of the small molecular weight polysaccharide in the rhizome atractylodes rootlet, and develop it into an anti-aging skincare product, which helps to improve the comprehensive utilization value of rhizoma atractylodis. SUMMARY

[0004] In view of this, the present application aims to provide a rhizoma atractylodis rootlet small molecular weight polysaccharide and its application in anti-aging skincare products.

[0005] In order to achieve the above-mentioned purpose, a first object of the present application is to provide a North Atractylodes rhizome root polysaccharide, which adopts the following technical scheme:

[0006] The North Atractylodes rhizome root polysaccharide is a type of inulin arabino-fructan, the main chain structure is →1-β-D-Fruf-2→1-β-D-Fruf-2→, the branched chain structure is α-L-Araf-(1→5)-α-L-Araf-(1→, and is connected with the main chain through O-6 of →1,6-β-D-Fruf-2→; and the molecular weight of the North Atractylodes rhizome root polysaccharide is 3.7 kD.

[0007] It is worth noting that the monosaccharide composition of the North Atractylodes rhizome root polysaccharide disclosed in the present application is mainly composed of fructose and arabinose, and only contains a small amount of galactose and glucose.

[0008] A second object of the present application is to provide the application of the North Atractylodes rhizome root polysaccharide as described above.

[0009] The application of the North Atractylodes rhizome root polysaccharide in the preparation of an anti-aging skin care product.

[0010] Further, the application of the North Atractylodes rhizome root polysaccharide in the preparation of a product for significantly promoting the proliferation of keratinocyte HaCAT.

[0011] Further, the application of the North Atractylodes rhizome root polysaccharide in the preparation of a product for inhibiting the activity of β-galactosidase in keratinocyte HaCAT.

[0012] Further, the application of the North Atractylodes rhizome root polysaccharide in the preparation of a product for inhibiting the generation of ROS active oxygen in keratinocyte HaCAT.

[0013] Further, the application of the North Atractylodes rhizome root polysaccharide in the preparation of a product for inhibiting the generation of ROS active oxygen in AB type zebra fish.

[0014] Further, the application of the North Atractylodes rhizome root polysaccharide in the preparation of a skin anti-aging cream product.

[0015] Further, the skin anti-aging cream product contains, by mass:

[0016] 1,3-propanediol 5-8 wt%, ethylenediaminetetraacetic acid 0.2-0.6 wt%, squalane 4-5 wt%, cetyl stearyl alcohol 1.0-1.5 wt%, dimethicone 1-1.2 wt%, isopropyl palmitate 2-2.5 wt%, hydroxyethyl acrylate / acryloyldimethyltaurate sodium copolymer 0.1-0.3 wt%, 1,2-pentanediol 0.5-0.7 wt%, phenoxyethanol 0.05-0.2 wt%, North Atractylodes rhizome root polysaccharide 1-5 wt%, and the balance of water.

[0017] Compared with the prior art, the present application discloses the specific structure of the polysaccharide with a molecular weight of 3.7 kD in the rhizome of Atractylodes chinensis, studies the corresponding functional properties, and provides the application of the polysaccharide with the specific structure in the skin anti-aging activity, which can promote the proliferation of HaCAT cells, inhibit the activity of beta-galactosidase in HaCAT cells and the generation of ROS active oxygen to improve the anti-aging activity of products. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced below. Obviously, the drawings in the following description are only embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor on the basis of the provided drawings.

[0019] Figure 1 The molecular weight of the purified rhizome polysaccharide of Atractylodes chinensis disclosed in Embodiment 2 of the present application.

[0020] Figure 2 The monosaccharide composition of the rhizome polysaccharide of Atractylodes chinensis disclosed in Embodiment 3 of the present application.

[0021] Figure 3 The nuclear magnetic resonance spectrum of the rhizome polysaccharide of Atractylodes chinensis disclosed in Embodiment 5 of the present application, wherein A to F are 1 H, 13 C, Dept135, COSY, HSQC and HMBC, and G is a structural schematic diagram of the rhizome polysaccharide of Atractylodes chinensis.

[0022] Figure 4 The experimental results of the rhizome polysaccharide of Atractylodes chinensis promoting HaCaT cell proliferation disclosed in Embodiment 6 of the present application, wherein A is the effect of the rhizome polysaccharide of Atractylodes chinensis on the activity of HaCaT cells; B is the positive cell rate of HaCaT cells under the Edu fluorescence staining of the rhizome polysaccharide of Atractylodes chinensis; and C is the Edu fluorescence staining diagram of HaCaT cells under the rhizome polysaccharide of Atractylodes chinensis.

[0023] Figure 5 The determination results of the beta-galactosidase activity in HaCAT cells disclosed in Embodiment 7 of the present application.

[0024] Figure 6 The determination results of the ROS content in HaCAT cells disclosed in Embodiment 8 of the present application.

[0025] Figure 7 The determination results of the ROS content in zebrafish disclosed in Embodiment 9 of the present application. DETAILED DESCRIPTION

[0026] The technical solutions in the embodiments of the present application will be clearly and completely described below, obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work belong to the protection scope of the present application.

[0027] Herein, the word "embodiment" as "exemplary" explained any embodiment does not necessarily explain as superior or better than other embodiments. The performance index test in the embodiments of the present application adopts the conventional test method in the art, unless otherwise specified. It should be understood that the terms described in the present application are only for describing the specific embodiments, and are not used to limit the disclosure of the present application.

[0028] Unless otherwise specified, the technical and scientific terms used herein have the same meanings as generally understood by those skilled in the art to which the present application belongs; as the test methods and technical means not specially noted in the present application are the experimental methods and technical means generally used by those skilled in the art.

[0029] In order to better illustrate the content of the present application, a large number of specific details are given in the specific embodiments below. Those skilled in the art should understand that without certain specific details, the present application can also be implemented. In the embodiments, some methods, means, instruments, equipment and the like which are well known to those skilled in the art are not described in detail, in order to highlight the main idea of the present application.

[0030] The technical features disclosed in the embodiments of the present application can be combined arbitrarily without conflict, and the technical solutions obtained belong to the disclosure of the embodiments of the present application.

[0031] The present application discloses a kind of atractylodes lancea rhizome root polysaccharide and application, belong to plant polysaccharide technical field.The specific structure of the polysaccharide with the molecular weight of 3.7kD in atractylodes lancea rhizome root is determined by the present application, for studying its corresponding functional properties, and the application of the polysaccharide of this specific structure in skin anti-aging activity is provided, can promote the proliferation of keratinocyte HaCAT, inhibit the activity of keratinocyte HaCAT β-galactosidase and ROS active oxygen generation to improve product anti-aging activity.

[0032] In order to better understand the present application, the present application is further specifically described by the following examples, but it should not be understood as limiting the present application. Some non-essential improvements and adjustments made by those skilled in the art based on the above disclosure are also considered to fall within the protection scope of the present application.

[0033] The experimental materials involved in the embodiments of the present application include:

[0034] (1) North atractylodes rhizome root by Jiamusi University Science and Technology Park Hua Hao detection;

[0035] (2) Reactive oxygen species (ROS) and beta-galactosidase kit provided by Biyun Tian Biotechnology Co., Ltd.;

[0036] (3) Wild AB type zebra fish provided by the National Zebra Fish Resource Center (Wuhan, China);

[0037] (4) HaCaT cell line provided by Dr. Bioengineering Co., Ltd. (Wuhan, China);

[0038] (5) Levodopa by Shanghai Yuan Ye Biotechnology Co., Ltd.

[0039] Example 1: Preparation of North atractylodes rhizome root polysaccharide

[0040] Extraction, separation and purification of North atractylodes rhizome root crude polysaccharide:

[0041] Step A, the North atractylodes rhizome is crushed, sieved to get powder, and then defatted with petroleum ether, dried in the air, then soaked in 10 times the amount of deionized water for 12h, extracted by water extraction method, and the extract was concentrated. Add 4 times the volume of anhydrous ethanol to the concentrated solution and precipitate overnight. Centrifuge at 4000 rpm for 10 minutes to get the precipitate. Remove protein by Sevage method, dialyze the solution, then freeze-dry to obtain crude polysaccharide.

[0042] Step B, the North atractylodes rhizome crude polysaccharide aqueous solution was loaded on DE-52 chromatography column for chromatography analysis, and different ion strength polysaccharides were eluted. The eluted fraction (15mL / tube) was collected by automatic collector, and then the content of polysaccharide in each tube fraction was determined by phenol sulfuric acid method, and the enzyme marker was detected at 490nm. Draw scatter plot. Collect the eluent in different eluent range, concentrate, dialyze in 3500Da dialysis bag, freeze-dry.

[0043] Step C, the polysaccharide in water part of the eluent was purified by automatic gel purification system (BRT-GS), combined with online detection of differential detector (SHIMADZU) to collect the eluent. The collected solution was dialyzed, concentrated by rotary evaporator, and freeze-dried to obtain gel separation and purification of North atractylodes rhizome polysaccharide.

[0044] Example 2: Determination of molecular weight of North atractylodes rhizome polysaccharide

[0045] The relative average molecular weight and purity were determined by high performance gel permeation chromatography (HPGPC) equipped with refractive index detector (RID) (RI-10A, Shimadzu, Japan), 105-103-101 series gel column (8.0mm x 300mm).

[0046] Step A, first chromatographic analysis was performed using eight different molecular weight dextran series standards (P5, P10, P20, P50, P100, P200, P400, P800) (SHOWA DENKO a, Japan).

[0047] Step B, chromatographic conditions: 0.05M NaCl as mobile phase, flow rate of 0.7 mL / min, temperature of 40°C, injection volume of 25 μL, and monitoring by differential refractive index (dRI), calibration curve was established at a concentration of 5 mg / mL, and molecular weight calculation formula was obtained.

[0048] Step C, similarly, 25 μL sample of the purified polysaccharide solution (5 mg / mL) was placed on HPGPC after treatment, and chromatographic analysis was performed using the same chromatographic conditions as the standards, chromatogram and retention time were obtained, and molecular weight results were obtained by substituting the retention time into the molecular weight calculation formula.

[0049] As shown in Figure 1 chromatogram results show that at a retention time of 41.365 min, the chromatographic peak presents a single and symmetrical sharp peak, Mw (weight average molecular weight) and Mn (number average molecular weight) are both 3.7 kDa, the ratio of polydispersity index Mw / Mn is about equal to 1, the purity is 94.56%, the distribution is concentrated, and the uniformity is verified.

[0050] Example 3: Monosaccharide composition analysis of North Atractylodes rhizome root polysaccharide

[0051] Step A: 5 mg of polysaccharide was dissolved in 2 mL of 3M TFA, and hydrolysis was performed at 80°C for 2 h.

[0052] Step B: The hydrolyzed sample was blown dry with nitrogen at 60°C, and the blown dry material was completely mixed and dissolved in 5 mL of deionized water by vortexing, and the mixed material was centrifuged at 12000 rpm for 5 minutes, 50 μL of supernatant was taken and added to 950 μL of deionized water, which was the sample to be tested.

[0053] Step C: Then, the sample was analyzed using HPAEC-PAD equipped with Dionex Carbopac™ PA20 (150 mm x 3.0 mm), the mobile phase eluent was composed of A: H2O; B: 15 mM NaOH; C: 15 mM NaOH and 100 mM NaAc, the flow rate was 0.3 mL / min, and the sample size was 25 μL.

[0054] Step D: The standard monosaccharide mixture comprising mannose, rhamnose, galacturonic acid, galactose, glucose, glucuronic acid, arabinose, xylose, fucose, glucosamine hydrochloride, N-acetyl-D-glucosamine, D-fructose, D-ribose, galactosamine hydrochloride, L-guluronic acid, and D-mannuronic acid was treated using the same derivatization and analytical methods as the test sample.

[0055] Step E: Identify the monosaccharide composition and molar ratio of the polysaccharide from the rootlets of Atractylodes lancea by comparing the retention times of the chromatographic peaks of the monosaccharide mixture and the sample polysaccharide.

[0056] like Figure 2 As shown, the monosaccharide composition is mainly composed of fructose and arabinose, with only small amounts of galactose and glucose.

[0057] Example 4: Methylation analysis of polysaccharides from the fibrous roots of Atractylodes lancea

[0058] Step A: Weigh 2-3 mg of Atractylodes lancea root polysaccharide sample and place it in a glass reaction flask. Add 1 mL of anhydrous DMSO, then quickly add methylation reagent A and seal the flask. Dissolve the sample under sonication, then add methylation reagent B. React in a magnetically stirred water bath at 30°C for 60 min.

[0059] Step B: Finally, add 2 mL of ultrapure water to the above mixture to terminate the methylation reaction, dialyze in a 1000 Da dialysis bag for 24 h, and then freeze dry.

[0060] Step C: Accurately weigh 2 mg of the dialysis sample and 200 mg of potassium bromide, compress them into tablets, and use potassium bromide powder tablets as blank controls. Scan and record the results using a Fourier transform infrared spectrometer (FT-IR650, Tianjin Gangdong Technology Development Co., Ltd.) to confirm complete methylation before proceeding with subsequent experiments.

[0061] Step D: Take the methylated polysaccharide, add 1 mL of 2M trifluoroacetic acid (TFA) to hydrolyze for 90 min, and then evaporate to dryness using a rotary evaporator.

[0062] Step E: Add 2 mL of double-distilled water to the residue, reduce with 60 mg of sodium borohydride for 8 hours, neutralize with glacial acetic acid, rotary evaporate, and dry in an oven at 101 °C.

[0063] Step F, then add 1 mL of acetic anhydride and react at 100 °C for 1 h, then cool.

[0064] Step G, then add 10 mL of pure water to terminate the reaction.

[0065] Step H: The acetylated product was dissolved in 3 mL of CH₂Cl₂ and transferred to a separatory funnel. A small amount of distilled water was added, and the mixture was shaken thoroughly. The supernatant was then removed. This process was repeated four times. The CH₂Cl₂ layer was dried over an appropriate amount of anhydrous sodium sulfate, concentrated to 1 mL, and transferred to a liquid chromatography vial. Analysis of the acetylated product sample was performed using a Thermo Scientific 1300-7000 gas chromatograph-mass spectrometer.

[0066] GC-MS testing conditions: HP-INNOWAX column 30m*0.32mm*0.25um; temperature program conditions: initial temperature 140℃, increased to 230℃ at 1℃ / min; injection port temperature 250℃, detector temperature 250℃, carrier gas helium, flow rate 1mL / min.

[0067] As shown in Table 1, the methylation analysis results indicate that the refined polysaccharide from the rootlets of Atractylodes lancea contains nine glycosidic bonds.

[0068] Table 1. Results of polysaccharide methylation analysis of Atractylodes lancea rootlets

[0069]

[0070] Example 5: Nuclear magnetic resonance spectroscopy analysis of polysaccharides from the fibrous roots of Atractylodes lancea

[0071] Step A: Weigh 50 mg of purified polysaccharide sample from the rootlets of Atractylodes lancea and dissolve it in 0.5 mL of D2O. Then, detect the sample using an AVANCE 600 NMR spectrometer (600 MHz, Bruker, Germany).

[0072] Step B, NMR spectroscopy includes 1 H, 13 C, Dept 135, HSQC, HMBC 1 H / 1 H COSY.

[0073] Step C: Analyze the data using the MestReNova software (Santiago de Compostela, Spain).

[0074] The results are as follows Figure 3 As shown, the polysaccharide from the fibrous roots of Atractylodes lancea ( Figure 3 The main chain of G is →1-β-D-Fruf-2→1-β-D-Fruf-2→, and the side chain is α-L-Araf-(1→5)-α-L-Araf-(1→ connected to the main chain via O-6 of →1,6-β-D-Fruf-2→.

[0075] Example 6: Effects of Atractylodes lancea root polysaccharide on HaCAT cell proliferation and viability

[0076] Cell proliferation and cell vitality are key factors in anti-aging. Maintaining and enhancing cell proliferation capacity, as well as preserving cell vitality, are crucial for slowing down the aging process.

[0077] (1) HaCAT cell proliferation assay

[0078] Step A: HaCaT cells were cultured in DMEM high glucose medium containing 10% fetal bovine serum and 1% antibiotics (penicillin, streptomycin) and placed in a humidified incubator at 37°C and 5% CO2.

[0079] Step B: Take cells in the logarithmic growth phase for the experiment, at a concentration of 3*102 3 HaCaT cells were seeded into 96-well cell culture plates at a density of 1 cell / well.

[0080] Step C: After the cells adhered, different doses of polysaccharide solution (200, 400, 800 μg / mL) and 1 mM H2O2 were added. A blank control group containing only the culture medium was also set up. Cells were cultured for 24 and 48 hours, respectively.

[0081] Step D: Add 20 μL of MTT solution to each well and continue incubation in an incubator for 4 hours.

[0082] Step E: Then add 200 μL of DMSO to each well to dissolve and shake for 10 minutes.

[0083] Step F: Measure the absorbance of each well at a wavelength of 490 nm using a microplate reader.

[0084] (2) Cell viability test

[0085] Step A: HaCaT cells were incubated with EdU reagent for 2 hours.

[0086] Step B: Fix with paraformaldehyde (4%) at room temperature for 30 minutes.

[0087] Step C: Then, use Triton X-100 (0.4%) to penetrate for 10 minutes.

[0088] Step D: Stain with 1×Apollo reagent (15 μL) at room temperature for 30 minutes.

[0089] Step E: Then stain with DAPI solution at room temperature for 10 minutes.

[0090] Step F: Observe the image using a Zeiss AxioImager Z1 fluorescence microscope (Carl Zeiss, Oberkochen, Germany).

[0091] Step G: Finally, use ImageJ software to analyze the number of Edu-positive cells.

[0092] like Figure 4 As shown, with the gradual increase of the concentration of polysaccharide from the rootlets of Atractylodes lancea, it promoted the proliferation of HaCAT and the gradual increase of EDU-positive cells in a dose-dependent manner, demonstrating that it has a significant effect on promoting cell proliferation.

[0093] Example 7: Assay of β-galactosidase activity in HaCAT cells

[0094] Increased cellular β-galactosidase activity is an important marker of cellular senescence, and regulating this enzyme activity may have potential significance in combating aging.

[0095] Step A, HaCaT cells were added at a rate of 3*10 3 A density of 1 cell / well was seeded in a 6-well plate.

[0096] Step B involves treating the sample with different doses of polysaccharide solution (200, 400, and 800 μg / mL) and 1 mM H2O2 for 48 hours.

[0097] Step C: Remove the culture medium and wash the cells with PBS.

[0098] Step D: Add 1 mL of β-galactosidase staining fixative and fix at room temperature for 15 minutes.

[0099] Step E: Remove the cell fixative and wash the cells with PBS.

[0100] Step F: Add 1 mL of staining working solution to each well and incubate at 37°C for 12 h.

[0101] Step G: Observe and count the cells under an optical microscope. The cells stained dark blue are senescent positive cells.

[0102] like Figure 5 As shown, the polysaccharide from the rootlets of Atractylodes lancea inhibited H2O2-induced β-galactosidase activity in HaCaT cells in a dose-dependent manner.

[0103] Example 8: Determination of intracellular ROS content in HaCAT cells

[0104] Excessive ROS in the body can cause cumulative damage to cells, which is one of the aging mechanisms proposed by the free radical theory. ROS can react with a variety of biological macromolecules in cells (such as DNA, proteins, and lipids), leading to changes in the function and damage of these macromolecules.

[0105] Step A, HaCaT cells were added at a rate of 1*10 5 Cells were seeded at a density of 10 cells / well into 6-well cell culture plates.

[0106] Step B involves treating the sample with different doses of polysaccharide solution (200, 400, and 800 μg / mL) and 1 mM H2O2 for 48 hours.

[0107] Step C: After 48 hours, HaCaT cells were collected and placed in 10 μM DCFH-DA solution diluted in serum-free medium and incubated at 37°C for 20 minutes.

[0108] Step D: Wash the cells three times with serum-free culture medium to thoroughly remove any DCFH-DA that has not entered the cells.

[0109] Step E: Use a fluorescence microscope to detect the fluorescence signal of ROS and use image-J software to quantify the fluorescence intensity.

[0110] like Figure 6 As shown, polysaccharides from the rootlets of Atractylodes lancea inhibited H2O2-induced intracellular ROS levels in HaCaT cells in a dose-dependent manner.

[0111] Example 9: Determination of ROS content in zebrafish

[0112] Excessive ROS in the body can cause cumulative damage to cells, which is one of the aging mechanisms proposed by the free radical theory. ROS can react with a variety of biological macromolecules in cells (such as DNA, proteins, and lipids), leading to changes in the function and damage of these macromolecules.

[0113] Step A: Zebrafish embryos are seeded into 6-well cell culture plates at a density of 30 per well.

[0114] Step B involves treating the sample with different doses of polysaccharide solution (200, 400, and 800 μg / mL) and 1 mM H2O2 for 96 hours.

[0115] Step C: After 96 hours, the cultured zebrafish were placed in a diluted 10 μM DCFH-DA solution and incubated at 37°C for 20 minutes.

[0116] Step D: Wash three times with deionized water to thoroughly remove any DCFH-DA that has not entered the cells.

[0117] Step E: Use a fluorescence microscope to detect the fluorescence signal of ROS and use image-J software to quantify the fluorescence intensity.

[0118] like Figure 7 As shown, polysaccharides from the rootlets of Atractylodes lancea inhibited H2O2-induced ROS levels in zebrafish in a dose-dependent manner.

[0119] Example 10: Anti-aging skin cream product and its effect experiment

[0120] The composition of anti-aging skin cream products is shown in Table 2.

[0121] Table 2 Ingredients of Cream Products

[0122]

[0123]

[0124] Human body testing experiments:

[0125] Step A: Using the VISIA testing instrument, 20 subjects aged 35-65 years were selected. All subjects exhibited symptoms such as dull skin, wrinkles, decreased collagen fiber elasticity, and dark brown spots. Furthermore, the subjects were in good health, had no history of skin diseases or allergies, had non-sensitive skin, and were able to use skincare products as prescribed.

[0126] Step B: Subjects applied the cream prepared in Example 9 to their faces twice a day, morning and evening, for 28 days, and compared their skin condition before and after the application.

[0127] Step C: Calculate the wrinkle reduction rate and skin elasticity improvement rate for each group of subjects.

[0128] Table 3 shows the results of human evaluation of anti-aging cream products, indicating that cream products containing polysaccharide components with a molecular weight of 3.7kD from Atractylodes macrocephala rootlets have significant anti-aging effects.

[0129] Table 3 Results of Human Assessment Experiment

[0130]

[0131] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The application of a polysaccharide from the rootlets of Atractylodes lancea in the preparation of anti-aging skincare products, characterized in that, The polysaccharide from the rootlets of Atractylodes lancea is an inulin-type arabinofructose with a main chain structure of →1-β-D-Fruf-2→1-β-D-Fruf-2→ and a branched chain structure of α-L-Araf-(1→5)-α-L-Araf-(1→), which is connected to the main chain through the O-6 of →1,6-β-D-Fruf-2→; and the molecular weight of the polysaccharide from the rootlets of Atractylodes lancea is 3.7 kDa.

2. The application according to claim 1, characterized in that, The application of the polysaccharide from the rootlets of Atractylodes lancea in the preparation of products that promote the proliferation of HaCAT cells in keratinocytes.

3. The application according to claim 1, characterized in that, The application of the polysaccharide from the rootlets of Atractylodes lancea in the preparation of a product that inhibits the activity of β-galactosidase in HaCAT cells of keratinocytes.

4. The application according to claim 1, characterized in that, The application of the polysaccharide from the rootlets of Atractylodes lancea in the preparation of a product that inhibits the generation of reactive oxygen species (ROS) in HaCAT keratinocytes.

5. The application according to claim 1, characterized in that, The application of the polysaccharide from the rootlets of Atractylodes lancea in the preparation of products that inhibit the generation of reactive oxygen species (ROS) in type AB zebrafish.

6. The application according to claim 1, characterized in that, The application of Atractylodes macrocephala root polysaccharide in the preparation of anti-aging skin cream products.

7. The application according to claim 6, characterized in that, By weight, the skin anti-aging cream product comprises: 1,3-propanediol 5-8wt%, ethylenediaminetetraacetic acid 0.2-0.6wt%, squalane 4-5wt%, cetearyl alcohol 1.0-1.5wt%, polydimethylsiloxane 1-1.2wt%, isopropyl palmitate 2-2.5wt%, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.1-0.3wt%, 1,2-pentanediol 0.5-0.7wt%, phenoxyethanol 0.05-0.2wt%, Atractylodes lancea root polysaccharide 1-5wt%, and the balance being water.

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