A cosmetic DNA sodium raw material and its whitening and sunscreen application and preparation method

Abstract

The invention discloses a cosmetic sodium DNA raw material and its whitening and sunscreen application and preparation method, and relates to the field of biotechnology. The invention provides a method for preparing sodium DNA, a cosmetic raw material, comprising using a lysate to lyse fish tissue, wherein the lysate comprises 1-10mol / L EDTA, 1-5mol / L NaCl, 0.1-3mmol / L urea, 0.01-2% sodium N-lauroyl sarcosinate by mass fraction, 0.1-2% Brij30 by mass fraction, 5-15mmol / L sodium citrate and 40-80mmol / L Tris-HCl, and the obtained sodium DNA has a high purity and yield. When sodium DNA is used to prepare cosmetics, it has a good effect of inhibiting tyrosinase activity and a sunscreen effect, and has a strong antioxidant capacity, and has broad application prospects in the field of cosmetics.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a cosmetic DNA sodium raw material and its whitening and sunscreen application and a preparation method. Background Art

[0002] Sodium DNA, also known as polydeoxyribonucleotide (PDRN), is a deoxyribonucleotide polymer and a small molecule linear deoxyribonucleotide fragment. It has no immunogenicity in the human body and does not cause allergic reactions. It can efficiently activate adenosine A2A receptors. Activated adenosine A2A receptors can inhibit the production of reactive oxygen species, while promoting the release of cytokines such as VEGF, angiogenin and glutamine transferase II, promoting the proliferation of fibroblasts and fat precursor cells, and thus promoting the healing of ulcer surface.

[0003] People's skin becomes darker due to the action of melanin, which is a biological pigment present in the basal layer of the skin. Melanin is synthesized in the melanosomes of melanocytes. There are two types of melanin: eumelanin and pheomelanin. The former is black or dark brown and does not contain sulfur atoms in its structure; the latter is reddish brown or yellow and contains sulfur atoms in its structure. The initial stage of melanin synthesis is the generation of dopaquinone through the action of tyrosinase and chemical reaction of tyrosine.

[0004] Direct exposure of the skin to ultraviolet rays will accelerate aging and cause a series of physiological damages. Ultraviolet rays have strong penetrating power, and long-term exposure can directly reach the dermis, causing a series of adverse reactions to the skin. Continuous exposure of the skin to ultraviolet rays will cause many disadvantages, such as sunburn, basal cell and squamous cell carcinoma, melanocytoma, etc., so certain defensive measures need to be taken. It is very important to provide a cosmetic that has a sunscreen effect and a certain whitening effect.

[0005] The prior art discloses a variety of methods for preparing polydeoxyribonucleotides. For example, CN111214694A discloses a dressing with hemostasis and wound healing acceleration functions, which contains small molecule polydeoxyribonucleotides extracted from salmon. The preparation method of the small molecule polydeoxyribonucleotide comprises the following steps: (1) lysing salmon tissue using an alkaline lysis solution, and bathing in a constant temperature water bath at 90-100° C. for 20-40 min; the alkaline lysis solution contains 1-3 mol / L NaOH, 50-200 mM NaCl, and 5-25 mg / ml proteinase K; (2) adding Tris-HCl to the reaction system obtained in step (1); (3) adding HCL to the reaction system obtained in step (2); (4) centrifuging the reaction system obtained in step (3) and collecting the supernatant; (5) adding isopropanol to the supernatant obtained in step (4) and leaving it for more than 30 min; (6) centrifuging the reaction system obtained in step (5) and retaining the precipitate to obtain the small molecule polydeoxyribonucleotide. However, the purity of PDRN prepared by the preparation method disclosed in the invention needs to be further improved.

[0006] Chinese patent CN106031709A discloses fish sperm DNA-NA, protamine extract and its preparation method, the method is as follows: (1) take fish spermatozoa, add water to wash, drain the water to obtain fish spermatozoa raw material; (2) add 1-10 times volume of water, add 0.1-1.2% of hydrolysis enzyme at a temperature of 37-58°C, and perform enzymolysis for 30-300 minutes; (3) concentrate the filtrate to 30-80%; (4) adjust the pH to 6.0-8.0, add a cation exchange resin column, collect the effluent, concentrate the combined effluent and eluate, ethanol precipitate, and collect the precipitate; (5) continue to elute the cation exchange resin with 1-5% sodium chloride solution until the effluent is protein-free, and collect the eluate. The yield and purity of the protamine extract and fish sperm DNA-NA extract are low.

[0007] At present, there is an urgent need for a method for preparing sodium DNA with high purity and yield of extracted sodium DNA, and the prepared sodium DNA has good whitening and sunscreen effects. Summary of the invention

[0008] The invention aims to provide a cosmetic sodium DNA raw material and its whitening and sunscreen application and preparation method.

[0009] To achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0010] On the one hand, the present invention provides a method for preparing sodium DNA, comprising lysing fish tissue using a lysing solution, wherein the lysing solution comprises 1-10 mol / L EDTA, 1-5 mol / L NaCl, 0.1-3 mmol / L urea, 0.01-2% sodium N-lauroyl sarcosinate by mass, 0.1-2% Brij30 by mass, 5-15 mmol / L sodium citrate and 40-80 mmol / L Tris-HCl.

[0011] Furthermore, the lysis solution includes 1-6 mol / L EDTA, 2-5 mol / L NaCl, 0.5-3 mmol / L urea, 0.05-1.5% sodium N-lauroyl sarcosinate by mass, 0.5-1.5% Brij30 by mass, 8-15 mmol / L sodium citrate and 50-70 mmol / L Tris-HCl.

[0012] Most preferably, the lysate comprises 1 mol / L EDTA, 2 mol / L NaCl, 0.5 mmol / L urea, 0.05% sodium N-lauroyl sarcosinate by mass, 0.8% Brij30 by mass, 10 mmol / L sodium citrate and 60 mmol / L Tris-HCl.

[0013] Specifically, the preparation method of DNA sodium comprises the following steps:

[0014] (1) Centrifuge the fish tissue, discard the supernatant, and collect the precipitate;

[0015] (2) adding a lysis solution to the precipitate collected in step (1), and then adding a biological enzyme to obtain a lysate;

[0016] (3) Centrifuge the lysate in step (2) and take the supernatant;

[0017] (4) adding sodium chloride solution to the supernatant obtained in step (3), mixing and centrifuging, and taking the supernatant;

[0018] (5) Add ethanol to the supernatant obtained in step (4), centrifuge, collect the precipitate, and dry to obtain sodium DNA.

[0019] Specifically, the fish tissues described in step (1) include fish testes, semen and eggs.

[0020] Furthermore, the fish tissue described in step (1) is fish semen.

[0021] Specifically, the fish in step (1) is selected from the family Salmonidae.

[0022] Furthermore, the fish described in step (1) includes salmon, trout, and trout.

[0023] Furthermore, the fish in step (1) is salmon.

[0024] Specifically, the centrifugation condition in step (1) is centrifugation at 2000-5000 rpm for 10-40 min.

[0025] Furthermore, the centrifugation condition in step (1) is centrifugation at 2500 rpm for 15 min.

[0026] Specifically, the temperature for adding the cleavage solution in step (2) is 50-70° C., and the reaction time is 20-40 min.

[0027] Furthermore, the temperature for adding the cleavage solution in step (2) is 60° C. and the reaction time is 30 min.

[0028] Specifically, before adding the biological enzyme in step (2), the temperature is lowered to 35-45° C. and the pH is adjusted to 7-8.

[0029] Furthermore, before adding the biological enzyme in step (2), the temperature is lowered to 37° C. and the pH is adjusted to 7.5.

[0030] Specifically, the time for adding the biological enzyme reaction in step (2) is 1-5 hours.

[0031] Furthermore, the time for adding the bio-enzyme reaction in step (2) is 2 hours.

[0032] Specifically, the biological enzymes described in step (2) are trypsin and papain.

[0033] Furthermore, the mass ratio of trypsin to papain in step (2) is (0.1-3):1.

[0034] Furthermore, the mass ratio of trypsin to papain in step (2) is (1-2.5):1.

[0035] Furthermore, the mass ratio of trypsin to papain in step (2) is 1.5:1.

[0036] Specifically, in step (2), the mass volume ratio of the precipitate to the lysate is 1:(1-8).

[0037] Furthermore, in step (2), the mass volume ratio of the precipitate to the lysate is 1:(4-6).

[0038] Furthermore, in step (2), the mass volume ratio of the precipitate to the lysate is 1:5.

[0039] Specifically, the centrifugation condition in step (3) is 10000-15000 rpm for 5-30 min.

[0040] Specifically, the centrifugation condition in step (3) is centrifugation at 12500 rpm for 10 min.

[0041] Specifically, in step (4), the volume ratio of the supernatant to the sodium chloride solution is 1:(1-5).

[0042] Furthermore, in step (4), the volume ratio of the supernatant to the sodium chloride solution is 1:1.

[0043] Specifically, the centrifugation condition in step (4) is 10000-15000 rpm for 10-50 min.

[0044] Furthermore, the centrifugation condition in step (4) is 12500 rpm for 20 min.

[0045] Specifically, the volume ratio of the supernatant to ethanol in step (5) is 1:(1-3).

[0046] Furthermore, in step (5), the volume ratio of the supernatant to ethanol is 1:1.

[0047] Specifically, the centrifugation condition in step (5) is 4000-8000 rpm for 5-20 min.

[0048] Furthermore, the centrifugation condition in step (5) is centrifugation at 6000 rpm for 10 min.

[0049] Specifically, the drying method in step (5) includes but is not limited to freeze drying, reduced pressure drying, and microwave drying.

[0050] In another aspect, the present invention provides DNA sodium prepared by the above preparation method.

[0051] In another aspect, the present invention provides the use of sodium DNA prepared by the above-mentioned preparation method in the preparation of whitening and sunscreen products.

[0052] Specifically, the products include medicines and cosmetics.

[0053] More specifically, the cosmetics can be conventional preparations such as lotion, toner, toner, essence, cream, facial mask, cleansing cream, facial cleanser, shampoo and hair conditioner, shaving cream, hair milk, hair gel, lipstick, sunscreen, hand cream, etc.

[0054] More specifically, the cosmetics may also include base materials and conventional auxiliary materials.

[0055] More specifically, the matrix raw materials include but are not limited to colloid raw materials, powdery raw materials, oil raw materials, wax raw materials, coagulants, and surfactants.

[0056] More specifically, the conventional auxiliary materials include but are not limited to antioxidants, adhesives, lubricants, moisturizers, preservatives, whitening agents, film agents, emulsifiers, and cosmetic nutritional additives.

[0057] Specifically, the present invention provides an essence containing sodium DNA, which includes: 1-5 parts by weight of sodium DNA, 1-10 parts by weight of mannitol, 1-5 parts by weight of hyaluronic acid, 1-5 parts by weight of niacinamide, 1-5 parts by weight of lecithin, 70-90 parts by volume of water, 1-6 parts by volume of glycerol and 0.1-2 parts by weight of benzoic acid.

[0058] According to some embodiments of the present invention, the essence comprises 2 parts by weight of sodium DNA, 6 parts by weight of mannitol, 2 parts by weight of hyaluronic acid, 1 part by weight of niacinamide, 1 part by weight of lecithin, 90 parts by volume of water, 3 parts by volume of glycerol and 0.5 parts by weight of benzoic acid.

[0059] Furthermore, the present invention also provides a method for preparing the essence, comprising the following steps:

[0060] (1) Mix glycerol, hyaluronic acid and sodium DNA to obtain a premix;

[0061] (2) Add water to the emulsification tank, increase the temperature, add the premix to the emulsification tank, add lecithin, mix well, cool down, add mannitol, niacinamide, and benzoic acid, filter, and obtain the essence.

[0062] Specifically, the medicine also includes a pharmaceutically acceptable carrier.

[0063] Further preferably, the pharmaceutically acceptable carrier is selected from one or more of an excipient, a stabilizer, a diluent, a binder, a preservative, and a lubricant.

[0064] Furthermore, the dosage form of the drug is drops, mixtures, tinctures, injections, tablets, powders, oral liquids, capsules, granules, ointments, suspensions, powders, emulsions, solutions, pellets, pills, buccal tablets, freeze-dried powder injections, gels, suppositories or aerosols.

[0065] The beneficial effects of the present invention are:

[0066] The invention provides a method for preparing sodium DNA, which is a raw material for cosmetics. The sodium DNA prepared by the preparation method of the invention has high purity and yield. When the sodium DNA prepared by the invention is used to prepare cosmetics, the obtained cosmetics have good tyrosinase activity inhibition and sunscreen effects, and have strong antioxidant capacity, and can alleviate the damaging effects of oxidative stress on the skin. The sodium DNA of the invention has broad application prospects in the field of cosmetics. DETAILED DESCRIPTION

[0067] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further explained in conjunction with specific embodiments below, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without creative work all belong to the protection scope of the present invention. In the following embodiments, unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same.

[0068] The fish used in the following examples was salmon.

[0069] Example 1 Preparation of DNA sodium

[0070] The preparation of DNA sodium includes the following steps:

[0071] (1) Centrifuge the fish semen at 4°C and 2500 rpm for 15 min, discard the supernatant, and collect the precipitate;

[0072] (2) adding lysis solution to the precipitate collected in step (1) and reacting at 60°C for 30 min, lowering the temperature to 37°C, adjusting the pH to 7.5, adding biological enzyme to react for 2 h, and obtaining a lysate;

[0073] (3) Centrifuge the lysate in step (2) at 12500 rpm for 10 min and collect the supernatant;

[0074] (4) adding saturated sodium chloride solution to the supernatant obtained in step (3), mixing, the volume ratio of supernatant to sodium chloride solution being 1:1, centrifuging at 12500 rpm for 20 min, and collecting the supernatant;

[0075] (5) Add an equal volume of anhydrous ethanol to the supernatant obtained in step (4), centrifuge at 6000 rpm for 10 min after 30 min, collect the precipitate, and freeze-dry the precipitate in vacuum to obtain sodium DNA.

[0076] The mass volume ratio of the precipitate to the lysate in step (2) is 1 g:5 ml.

[0077] The lysis solution in step (2) contains 1 mol / L EDTA, 2 mol / L NaCl, 0.5 mmol / L urea, 0.05% sodium N-lauroyl sarcosinate by mass, 0.8% Brij30 by mass, 10 mmol / L sodium citrate and 60 mmol / L Tris-HCl.

[0078] The biological enzymes described in step (2) are trypsin and papain; the mass ratio of trypsin to papain is 1.5:1.

[0079] Example 2 Preparation of DNA Sodium

[0080] The preparation of DNA sodium includes the following steps:

[0081] (1) Centrifuge the fish semen at 4°C, 3000 rpm for 10 min, discard the supernatant, and collect the precipitate;

[0082] (2) adding lysis solution to the precipitate collected in step (1) and reacting at 60°C for 30 min, lowering the temperature to 37°C, adjusting the pH to 7.0, adding biological enzyme to react for 3 h, and obtaining a lysate;

[0083] (3) Centrifuge the lysate in step (2) at 12500 rpm for 10 min and collect the supernatant;

[0084] (4) adding saturated sodium chloride solution to the supernatant obtained in step (3), mixing, the volume ratio of supernatant to sodium chloride solution being 1:2, centrifuging at 12500 rpm for 20 min, and collecting the supernatant;

[0085] (5) Add 2 times the volume of anhydrous ethanol to the supernatant obtained in step (4), centrifuge at 6000 rpm for 10 min after 30 min, collect the precipitate, and freeze-dry the precipitate in vacuum to obtain sodium DNA.

[0086] The mass volume ratio of the precipitate to the lysate in step (2) is 1 g:6 ml.

[0087] The lysis solution in step (2) contains 2 mol / L EDTA, 2 mol / L NaCl, 1 mmol / L urea, 0.1% sodium N-lauroyl sarcosinate, 0.5% Brij30, 8 mmol / L sodium citrate and 50 mmol / L Tris-HCl.

[0088] The biological enzymes described in step (2) are trypsin and papain; the mass ratio of trypsin to papain is 1:1.

[0089] Example 3 Preparation of DNA sodium

[0090] The preparation of DNA sodium includes the following steps:

[0091] (1) Centrifuge the fish semen at 4°C and 3500 rpm for 10 min, discard the supernatant, and collect the precipitate;

[0092] (2) adding lysis solution to the precipitate collected in step (1) and reacting at 60°C for 30 min, lowering the temperature to 37°C, adjusting the pH to 7.0, adding biological enzyme to react for 2 h, and obtaining a lysate;

[0093] (3) Centrifuge the lysate in step (2) at 12,000 rpm for 15 min and collect the supernatant;

[0094] (4) adding saturated sodium chloride solution to the supernatant obtained in step (3), mixing, the volume ratio of supernatant to sodium chloride solution being 1:3, centrifuging at 12500 rpm for 20 min, and taking the supernatant;

[0095] (5) Add an equal volume of anhydrous ethanol to the supernatant obtained in step (4), centrifuge at 6000 rpm for 10 min after 30 min, collect the precipitate, and freeze-dry the precipitate in vacuum to obtain sodium DNA.

[0096] The mass volume ratio of the precipitate to the lysate in step (2) is 1 g:4 ml.

[0097] The lysis solution in step (2) contains 5 mol / L EDTA, 3 mol / L NaCl, 2 mmol / L urea, 1% sodium N-lauroyl sarcosinate, 1% Brij30, 10 mmol / L sodium citrate and 60 mmol / L Tris-HCl.

[0098] The biological enzymes described in step (2) are trypsin and papain; the mass ratio of trypsin to papain is 2:1.

[0099] Example 4 Preparation of DNA Sodium

[0100] The preparation of DNA sodium includes the following steps:

[0101] (1) Centrifuge the fish semen at 4°C and 3000 rpm for 15 min, discard the supernatant, and collect the precipitate;

[0102] (2) adding lysis solution to the precipitate collected in step (1) and reacting at 60°C for 30 min, lowering the temperature to 37°C, adjusting the pH to 7.0, adding biological enzyme to react for 2.5 h, and obtaining a lysate;

[0103] (3) Centrifuge the lysate in step (2) at 12500 rpm for 20 min and collect the supernatant;

[0104] (4) adding saturated sodium chloride solution to the supernatant obtained in step (3), mixing, the volume ratio of supernatant to sodium chloride solution being 1:2, centrifuging at 12500 rpm for 20 min, and collecting the supernatant;

[0105] (5) Add an equal volume of anhydrous ethanol to the supernatant obtained in step (4), centrifuge at 6000 rpm for 10 min after 30 min, collect the precipitate, and freeze-dry the precipitate in vacuum to obtain sodium DNA.

[0106] The mass volume ratio of the precipitate to the lysate in step (2) is 1 g:6 ml.

[0107] The lysis solution in step (2) contains 6 mol / L EDTA, 5 mol / L NaCl, 3 mmol / L urea, 1.5% sodium N-lauroyl sarcosinate, 1.5% Brij30, 15 mmol / L sodium citrate and 70 mmol / L Tris-HCl.

[0108] The biological enzymes described in step (2) are trypsin and papain; the mass ratio of trypsin to papain is 2.5:1.

[0109] Comparative Example 1 Preparation of DNA Sodium

[0110] The difference from Example 1 is that the contents of the components in the lysate in step (2) are different, and the rest are the same. The lysate in Comparative Example 1 contains 12 mol / L EDTA, 6 mol / L NaCl, 3.5 mmol / L urea, 2.5% sodium N-lauroyl sarcosinate, 2.5% Brij30, 20 mmol / L sodium citrate and 90 mmol / L Tris-HCl.

[0111] Comparative Example 2 Preparation of DNA Sodium

[0112] The difference from Example 1 is that the components of the lysate in step (2) are different, and the rest are the same. The lysate in Comparative Example 2 contains 1 mol / L EDTA, 2 mol / L NaCl, 0.5 mmol / L urea, 0.05% TritonX-100, 0.8% sodium dodecyl sulfate, 10 mmol / L sodium citrate, and 60 mmol / L Tris-HCl.

[0113] Comparative Example 3 Preparation of DNA Sodium

[0114] The only difference from Example 1 is that the mass volume ratio of the precipitate to the lysate in step (2) is 1 g:10 ml, and the rest is the same.

[0115] Comparative Example 4 Preparation of DNA Sodium

[0116] The only difference from Example 1 is that the biological enzymes in step (2) are proteinase K and alkaline protease, and the rest are the same.

[0117] Comparative Example 5 Preparation of DNA Sodium

[0118] The only difference from Example 1 is that the mass ratio of trypsin to papain in step (2) is 4:1, and the rest are the same.

[0119] Example 5 Preparation of DNA sodium-containing serum

[0120] The essence in this embodiment includes the following components: 2 parts by weight of sodium DNA, 6 parts by weight of mannitol, 2 parts by weight of hyaluronic acid, 1 part by weight of niacinamide, 1 part by weight of lecithin, 90 parts by volume of water, 3 parts by volume of glycerol and 0.5 parts by weight of benzoic acid.

[0121] The preparation of the serum comprises the following steps:

[0122] (1) Mix glycerol, hyaluronic acid and sodium DNA to obtain a premix;

[0123] (2) Add water to the emulsification tank, raise the temperature to 80°C, add the premix to the emulsification tank, add lecithin, mix well, cool to 40°C, add mannitol, niacinamide, and benzoic acid, filter through a 200-mesh filter to obtain an essence.

[0124] Experimental Example 1 Purity and Yield of DNA Sodium

[0125] Yield = product weight / testis weight;

[0126] The products obtained in the examples and comparative examples were dissolved in physiological saline and the absorbance at 260 nm and 280 nm was measured using a spectrophotometer. The purity of DNA sodium in the products was calculated according to the following formula:

[0127] Purity = OD 260 / OD 280 .

[0128] Table 1 Purity and yield

[0129]

[0130] Experimental Example 2 Tyrosinase Activity Inhibition Experiment

[0131] Mouse B16 melanocytes were cultured at a rate of 1×105 / ml were inoculated in a 96-well plate and cultured at 37°C and 5% CO2 for 24 hours. After discarding the supernatant, the essence containing the DNA sodium of Examples 1-4 and Comparative Examples 1-5 was added (the preparation methods of the essences of Examples 1-4 and Comparative Examples 1-5 were the same as those of Example 5, and the only difference was that the DNA sodium added was the DNA sodium prepared in Examples 1-4 and Comparative Examples 1-5, respectively). After 3 days, the supernatant was discarded, the plates were rinsed with PBS, 90 μL of PBS containing 1% TritonX-100 by volume was added to each well, and the plates were ultrasonically broken in an ice bath, and 10 μL of 10 mmol / L L-DOPA was added to each well. The plates were incubated at 37°C for 60 minutes, and the absorbance of each well was measured by colorimetry at 475 nm.

[0132] Inhibition rate (%) = (1-average absorbance value of drug group / average absorbance value of control group) × 100%.

[0133] Experimental results:

[0134] Table 2 Tyrosinase activity inhibition rate

[0135]

[0136] It can be seen from the above table that the DNA sodium prepared in Examples 1-4 of the present invention has a good effect of inhibiting tyrosinase activity, and the effect of the essence prepared in Comparative Examples 1-5 is significantly worse than that of Examples 1-4.

[0137] Experimental Example 3 Sunscreen Effect Testing

[0138] Before the experiment, a blank quartz plate was used to detect the background curve, and 0.2 mg / cm 2 The essence (the preparation methods of the essences of Examples 1-4 and Comparative Examples 1-5 are the same as those of Example 5, the only difference being that the added sodium DNA is the sodium DNA prepared in Examples 1-4 and Comparative Examples 1-5, respectively) is spread evenly on the surface of a 3 m tape, left to stand for 20 min, and tested using an ultraviolet transmittance analyzer (SPF-290AS, purchased from Solarlight) to obtain the UVB protection value, expressed as SPF.

[0139] SPF = minimum erythema dose (after using sunscreen) / minimum erythema dose (before using sunscreen).

[0140] Experimental results:

[0141] Table 3 SPF value

[0142]

[0143] Experimental Example 4 Antioxidant Capacity

[0144] Experimental methods:

[0145] HaCat cells were inoculated in 96-well plates and divided into a normal group, a H2O2 group, an Example 1 group, and a Comparative Example 1-5 group. Three replicates were set for each group. DMEM high-glucose medium (containing 10% fetal bovine serum, 100U / ml penicillin, and 100U / ml streptomycin) was added and cultured at 37°C and 5% CO2 for 12 hours. Except for the normal group, 150mM H2O2 was added to the other groups. The concentration of PDRN in the Example 1 group and the Comparative Example 1-5 groups was 100μg / ml. The antioxidant capacity of the cells was detected using the superoxide dismutase (SOD) detection kit and the malondialdehyde (MDA) detection kit of Nanjing Jiancheng Biotechnology Co., Ltd.

[0146] Experimental results:

[0147] Table 4 Antioxidant capacity test results

[0148]

[0149] Note: Compared with the normal group, * p <0.05,** p <0.01; compared with the H2O2 group, # p <0.05,## p <0.01.

[0150] It can be seen from the above table that the DNA sodium prepared in Example 1 of the present invention can effectively increase the antioxidant properties of HaCat cells, increase the activity of SOD and reduce the content of MDA. Although the DNA sodium prepared in Comparative Examples 1-5 can increase the activity of SOD and reduce the content of MDA, there is no significant difference compared with the H2O2 group.

[0151] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing DNA sodium, characterized in that: The preparation method comprises the following steps: (1) Centrifuge the fish tissue, discard the supernatant, and collect the precipitate; (2) adding a lysis solution to the precipitate collected in step (1), and then adding a biological enzyme to obtain a lysate; (3) Centrifuge the lysate in step (2) and take the supernatant; (4) adding sodium chloride solution to the supernatant obtained in step (3), mixing and centrifuging, and taking the supernatant; (5) adding ethanol to the supernatant obtained in step (4), centrifuging, collecting the precipitate, and drying to obtain sodium DNA; The lysate is composed of 1-10 mol / L EDTA, 1-5 mol / L NaCl, 0.1-3 mmol / L urea, 0.01-2% sodium N-lauroyl sarcosinate, 0.1-2% Brij30, 5-15 mmol / L sodium citrate and 40-80 mmol / L Tris-HCl; The biological enzymes described in step (2) are trypsin and papain; The mass ratio of trypsin to papain described in step (2) is (0.1-3):

1.

2. The preparation method according to claim 1, characterized in that: The lysate comprises 1 mol / L EDTA, 2 mol / L NaCl, 0.5 mmol / L urea, 0.05% sodium N-lauroyl sarcosinate by mass fraction, 0.8% Brij30 by mass fraction, 10 mmol / L sodium citrate and 60 mmol / L Tris-HCl.

3. The preparation method according to claim 1, characterized in that: The fish tissues described in step (1) include fish testes, semen and eggs.

4. The preparation method according to claim 1, characterized in that: The temperature for the reaction of adding the lysis solution in step (2) is 50-70°C, and the reaction time is 20-40 minutes.

5. The preparation method according to claim 1, characterized in that: The mass ratio of trypsin to papain in step (2) is 1.5:

1.

6. The preparation method according to claim 1, characterized in that: The mass volume ratio of the precipitate to the lysate in step (2) is 1:(1-8).

7. The preparation method according to claim 1, characterized in that: In step (4), the volume ratio of the supernatant to the sodium chloride solution is 1:(1-5).

8. The preparation method according to claim 1, characterized in that: In step (5), the volume ratio of the supernatant to ethanol is 1:(1-3).

9. DNA sodium prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the sodium DNA prepared by the preparation method according to any one of claims 1 to 8 in the preparation of whitening and sunscreen products.

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