Application of Tricarpellate Wampee Fruit Extract in Preparing External Preparation for Repairing Skin after Sunburn

Abstract

The present invention belongs to the technical field of cosmetics, and specifically relates to the application of the extract of Sparganium stoloniferum in the preparation of an external preparation for repairing skin after sunburn. The extract of Sparganium stoloniferum prepared by the present invention can scavenge free radicals, inhibit lipid peroxidation, inhibit the production of ROS, down-regulate the expression of AGEs, and can also inhibit the inflammatory response induced by ultraviolet rays, reduce the expression of NF-κB and HMGB1 genes up-regulated by ultraviolet rays, inhibit the secretion of the inflammatory factor TNF-α. At the same time, it can reduce the expression of the melanocyte-stimulating factor α-MSH and β-catenin genes up-regulated by ultraviolet rays, inhibit the production of melanin and the transport of melanosomes, that is, significantly reduce the symptoms of skin damage after sunburn by improving various signal pathways such as skin oxidative damage, inflammatory response, and pigmentation caused by ultraviolet rays. When it is applied to external preparations such as cosmetics for repairing skin damage after sunburn, it will have broad market application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and particularly relates to the application of a triangular prism extract in the preparation of an external preparation for skin repair after sunburn. Background Art

[0002] As an important organ in direct contact with the external environment, the skin is easily damaged by ultraviolet radiation (UVR) when exposed to solar radiation for a long time. According to the degree and duration of the damage, it is mainly divided into chronic damage and acute damage. Chronic damage, also known as photoaging, is mainly manifested as dryness, yellowing, relaxation, telangiectasia, a large number of deep wrinkles except fine lines, irregular pigmentation, etc.; according to the different degrees of damage, acute post-sun damage can be divided into mild pigmentation and severe sunburn. According to different mechanisms, skin pigmentation can be divided into immediate pigment darkening (IPD), persistent pigment darkening (PPD), and delayed pigment darkening (DT). Sunburn is mainly caused by excessive exposure of the skin to UVB irradiation, and its clinical manifestations are mainly erythema, stratum corneum desquamation, and edema, etc., accompanied by pain.

[0003] Further studies have shown that acute and chronic post-sun damage have the same inducing factors, which are specifically as follows:

[0004] (1) Light can activate the chromophores in the skin to react with molecular oxygen, resulting in the production of reactive oxygen species (ROS). When the generated ROS exceeds the body's own ability to scavenge ROS, oxidative stress occurs in the skin. Oxidative stress can lead to lipid, protein, and DNA damage, and even accelerate the formation of advanced glycation end products (AGEs) and further promote the inflammatory response.

[0005] (2) Light will damage the viable cell layer and skin barrier. The damaged cell membrane releases pro-inflammatory factors, which will further activate inflammation-related signaling pathways such as NF-κB, mediate the occurrence and intensification of the inflammatory response, express more inflammatory factors, and produce inflammatory infiltration. When post-sun damage occurs to the skin, the ability of the skin barrier to resist ultraviolet rays decreases, the immune response is inhibited, and HMGB1 is released from damaged cells or dead cells. As a signaling molecule, it participates in the inflammatory response and regulates the immune response, further strengthening the inflammatory response. Subsequently, symptoms of post-sun damage such as skin sunburn erythema, chronic inflammatory photoaging, skin sunburn darkening, and photosensitivity appear on the skin surface.

[0006] (3) Light can oxidize the melanin already present in cells by reactive oxygen species and other groups generated by UVA, turning it into oxidized melanin with a darker color and accelerating the transport of melanosomes within melanocytes. At the same time, after UVB irradiation stimulates the skin, by secreting related inducing factors such as the MC1R / α-MSH signaling pathway, the Wnt / β-catenin signaling pathway, etc., it further promotes the expression of MITF, activates tyrosinase within melanocytes, and ultimately leads to an increase in the number and activity of melanocytes to promote melanin synthesis; it causes the dendrites of melanocytes to grow, and at the same time stimulates mature melanosomes to transfer along the dendrites of melanocytes to keratinocytes, thus resulting in the skin tanning reaction.

[0007] There are relatively few functional preparations for repairing sun-exposed skin on the market currently. The research on the mechanism targets is not transparent enough, the mechanism of action is single and the effect is limited, and it is not sufficient to comprehensively solve the problem of sun-induced skin damage. It is necessary to stack multiple ingredients to achieve a certain effect. Summary of the Invention

[0008] The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide the application of Sparganium stoloniferum extract in the preparation of an external preparation for repairing sun-exposed skin.

[0009] In the first aspect, the present invention provides the application of Sparganium stoloniferum extract in the preparation of an external preparation for repairing sun-exposed skin.

[0010] Preferably, the Sparganium stoloniferum extract exerts the effect of repairing sun-exposed skin through at least one of the following aspects:

[0011] a. Inhibiting the oxidative stress generated by sun-induced damage to skin tissue;

[0012] b. Inhibiting the inflammatory response generated by sun-induced damage to skin tissue;

[0013] c. Inhibiting the pigmentation generated by sun-induced damage to skin tissue.

[0014] Sparganium stoloniferum is the dried tuber of Sparganium stoloniferum Buch.-Ham. ex Juz. of the Sparganiaceae family, with a bitter taste and a neutral nature, entering the liver and spleen meridians. It was first recorded in "Supplementary Records of Materia Medica" and has the effects of promoting blood circulation to remove blood stasis, promoting qi movement, and relieving accumulation and pain. It is used for mass in the abdomen, stasis of blood in menstruation, abdominal pain due to food accumulation, etc., and is a commonly used drug for promoting blood circulation and removing blood stasis in clinical practice. In the prior art, regarding the research on the application of Sparganium stoloniferum extract in the field of cosmetics, due to its good antibacterial effect, it is often compounded with other components for preventing or treating acne-related skin problems. Currently, there is no report on using Sparganium stoloniferum extract to improve sun-induced skin damage. Therefore, the functional research on using Sparganium stoloniferum extract to improve sun-induced skin damage has novelty and great market potential.

[0015] Through experimental exploration, it is found that the extract of Sparganium stoloniferum can protect the skin from post - sun damage caused by ultraviolet radiation by reducing multiple signaling pathways such as oxidative stress, inflammatory response, and pigmentation. Among them, the extract of Sparganium stoloniferum can improve cellular oxidative damage by scavenging free radicals, inhibiting lipid peroxidation, inhibiting the production of ROS, and down - regulating the expression of AGEs; the extract of Sparganium stoloniferum can also inhibit the inflammatory response induced by ultraviolet rays by reducing the expression of NF - κB and HMGB1 genes up - regulated by ultraviolet rays and inhibiting the secretion of the inflammatory factor TNF - α; in addition, the extract of Sparganium stoloniferum can also reduce the expression of the melanocyte - stimulating factor α - MSH and the synthesis and transport - related gene β - catenin up - regulated by ultraviolet rays, inhibit the production of melanin and the transport of melanosomes, thereby improving pigmentation caused by ultraviolet rays.

[0016] In a second aspect, the present invention provides a preparation method of the above - mentioned extract of Sparganium stoloniferum, comprising the following steps:

[0017] (1) Crush the Sparganium stoloniferum tubers, sieve through a 40 - 60 - mesh sieve, add an ethanol solution for soaking, then reflux for 1.5 - 2.5 h, filter to obtain the first extract, and then use the ethanol solution to reflux the filter residue for 1.5 - 2.5 h, filter to obtain the second extract;

[0018] (2) Then use the ethanol solution to reflux the filter residue for 0.5 - 1.5 h to obtain the third extract, combine the three extracts, remove ethanol under reduced pressure, filter, and take the filtrate to obtain the extract of Sparganium stoloniferum.

[0019] Preferably, the temperature of the three - time reflux extraction is 75 - 85 °C.

[0020] Preferably, the volume concentration of the ethanol solution is 60 - 70%.

[0021] The present invention uses the above - mentioned preparation method to extract the extract of Sparganium stoloniferum. The preparation process is simple. Through experimental exploration, it is found that the particle size of Sparganium stoloniferum granules, the reflux extraction temperature, the ethanol concentration used for extraction, and the extraction times will have an important impact on the content of flavonoid and polyphenol active ingredients in the finally obtained extract of Sparganium stoloniferum. Within the scope of the above - defined process conditions, it can effectively ensure the full dissolution of flavonoid and polyphenol active ingredients in Sparganium stoloniferum, and avoid their degradation or destruction, maximizing the extraction efficiency of active ingredients, so as to achieve a better post - sun damage repair effect.

[0022] In a third aspect, the present invention provides a skin post - sun repair cosmetic, the cosmetic comprising the extract of Sparganium stoloniferum prepared by the above - mentioned preparation method, and the mass percentage content of the extract of Sparganium stoloniferum in the cosmetic is 1 - 60%.

[0023] Preferably, the cosmetic further comprises common cosmetic adjuvants, including polyhydric alcohols, preservatives and solubilizers; the polyhydric alcohols include at least one of propylene glycol, pentylene glycol and dipropylene glycol; the preservatives include at least one of p-hydroxyacetophenone, chlorphenesin and phenoxyethanol; the solubilizers include at least one of PEG caprylic / capric glycerides and hydrogenated lecithin.

[0024] Preferably, the cosmetic dosage form includes at least one of aqueous solutions, emulsions and creams.

[0025] In a fourth aspect, the present invention provides an emulsion for repairing skin after sunburn, which comprises the rhizoma sparganii extract prepared by the above preparation method; the mass percentage content of the rhizoma sparganii extract in the emulsion is 1-3%.

[0026] Preferably, the emulsion further comprises 10-15% of a humectant, 5-10% of an emulsifier, 0.2-0.7% of an antioxidant, 1-3% of a thickener, 1-2% of a preservative and the balance of water.

[0027] Preferably, the humectant includes at least one of glycerol, sodium hyaluronate, propylene glycol, squalane, caprylic / capric glyceride, glucose, butanediol, 1,2-pentanediol, octyldecanol and isopentyl glycol;

[0028] The emulsifier includes at least one of acrylate / acrylamide copolymer, isopropyl myristate, C14-22 alcohol, C12-20 alkyl glucoside, stearic acid, cetearyl olivate, sorbitan olivate, span 80, glyceryl stearate, PEG-100 stearate, steareth-2, steareth-21, ceteth-12, ceteth-30, potassium cetyl phosphate, sodium stearoyl glutamate, polysorbate-60 and sorbitan stearate;

[0029] The antioxidant includes at least one of tocopherol and tocopheryl acetate;

[0030] The thickener includes at least one of acrylate crosspolymer-6, xanthan gum, cetearyl alcohol, carbomer and hydroxyethyl cellulose;

[0031] The preservative includes at least one of p-hydroxyacetophenone, 1,2-hexanediol, octanoyl hydroxamic acid, benzyl alcohol, phenoxyethanol, benzoic acid and its salts, sorbic acid and its salts.

[0032] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0033] The extracts of Sparganium stoloniferum prepared by the present invention can scavenge free radicals, inhibit lipid peroxidation, inhibit the production of ROS, down-regulate the expression of AGEs, and also inhibit the inflammatory response induced by ultraviolet rays, reduce the expression of NF-κB and HMGB1 genes up-regulated by ultraviolet rays, inhibit the secretion of inflammatory factor TNF-α. At the same time, it can reduce the expression of pro-melanogenic factors α-MSH and β-catenin genes up-regulated by ultraviolet rays, inhibit the production of melanin and the transport of melanosomes, that is, significantly reduce the symptoms of skin damage after sun exposure by improving various signaling pathways such as skin oxidative damage, inflammatory response, and pigmentation caused by ultraviolet rays. When it is applied to external preparations such as cosmetics for repairing skin damage after sun exposure, it will have broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It is the effect diagram of ROS scavenging. Among them, Figure a shows the ROS scavenging situation of the blank control group, Figure b shows the ROS scavenging situation of the positive control group (glutathione), and Figure c shows the ROS scavenging situation of 0.5% Sparganium stoloniferum extract;

[0035] Figure 2 It is the fluorescence microscope observation diagram of melanosomes after being treated with different test samples;

[0036] Figure 3 It is the relative average value (x±s) of melanosomes after being treated with different test samples. * indicates that the difference compared with M is statistically significant, **P<0.01;

[0037] Figure 4 It is the comparison diagram of skin pigmentation of the subjects before (a) and 56 days after (b) using the emulsion containing Sparganium stoloniferum extract;

[0038] Figure 5 It is the comparison diagram of ultraviolet-induced skin pigmentation of the subjects before (a) and 56 days after (b) using the emulsion containing Sparganium stoloniferum extract;

[0039] Figure 6 It is the comparison diagram of brown spots of the subjects before (a) and 56 days after (b) using the emulsion containing Sparganium stoloniferum extract;

[0040] Figure 7 It is the comparison diagram of red areas of the subjects before (a) and 56 days after (b) using the emulsion containing Sparganium stoloniferum extract. DETAILED DESCRIPTION OF THE INVENTION

[0041] To better illustrate the content of the present invention, the present invention will be further described below in conjunction with the drawings and specific embodiments.

[0042] Unless otherwise specified, the raw materials or reagents used in the present invention are conventional reagents existing in the art and can be obtained through commercial channels. The experimental operations not specifically described in the present invention are all conventional operations in the art or operations that can be understood or known by those skilled in the art according to the existing technology or common general knowledge they have mastered.

[0043] Example 1

[0044] This example provides a preparation method of a Sparganium extract, comprising the following steps:

[0045] Select the dry rhizomes of Sparganium stoloniferum Buch.-Ham. from the Sparganiaceae plant, pulverize them, sieve through a 50-mesh sieve, weigh 1 kg of Sparganium powder, add 5 times the amount of 70% ethanol, soak overnight, then reflux extract at 80 °C for 2 h, and filter; add 5 times the amount of 70% ethanol to the filter residue, reflux extract at 80 °C for 2 h and then filter; add 3 times the amount of 70% ethanol to the filter residue, reflux extract at 80 °C for 1 h, combine the three filtrates, recover ethanol under reduced pressure, let it stand and filter to obtain the said Sparganium extract.

[0046] Example 2

[0047] This example provides a preparation method of a Sparganium extract, comprising the following steps:

[0048] Select the dry rhizomes of Sparganium from the Sparganiaceae plant, pulverize them, sieve through a 40-mesh sieve, weigh 1 kg of Sparganium powder, add 5 times the amount of 60% ethanol, soak overnight, then reflux extract at 75 °C for 1.5 h, and filter; add 5 times the amount of 60% ethanol to the filter residue, reflux extract at 85 °C for 2 h and then filter; add 3 times the amount of 70% ethanol to the filter residue, reflux extract at 80 °C for 0.5 h, combine the three filtrates, recover ethanol under reduced pressure, let it stand and filter to obtain the said Sparganium extract.

[0049] Example 3

[0050] This example provides a preparation method of a Sparganium extract, comprising the following steps:

[0051] Select the dry rhizomes of Sparganium from the Sparganiaceae plant, pulverize them, sieve through a 60-mesh sieve, weigh 1 kg of Sparganium powder, add 5 times the amount of 70% ethanol, soak overnight, then reflux extract at 85 °C for 2.5 h, and filter; add 5 times the amount of 70% ethanol to the filter residue, reflux extract at 75 °C for 2 h and then filter; add 3 times the amount of 60% ethanol to the filter residue, reflux extract at 80 °C for 1.5 h, combine the three filtrates, recover ethanol under reduced pressure, let it stand and filter to obtain the said Sparganium extract.

[0052] Comparative Example 1

[0053] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: after the Sparganii Rhizoma tuber is crushed, it is sieved through a 20-mesh sieve, and the other conditions are the same.

[0054] Comparative Example 2

[0055] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: after the Sparganii Rhizoma tuber is crushed, it is sieved through an 80-mesh sieve, and the other conditions are the same.

[0056] Comparative Example 3

[0057] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: a conventional ultrasonic extraction method is used to replace the ethanol reflux extraction, and the conditions of the ultrasonic extraction are: ultrasonic extraction for 2 h at a power of 300 w and a temperature of 80 °C.

[0058] Comparative Example 4

[0059] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: the volume concentration of the ethanol solution is 50%, and the other conditions are the same.

[0060] Comparative Example 5

[0061] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: the volume concentration of the ethanol solution is 80%, and the other conditions are the same.

[0062] Comparative Example 6

[0063] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: the temperature during reflux extraction is 65 °C, and the other conditions are the same.

[0064] Comparative Example 7

[0065] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: the temperature during reflux extraction is 95 °C, and the other conditions are the same.

[0066] Comparative Example 8

[0067] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: the time for the third reflux extraction is 2 h, and the other conditions are the same.

[0068] Comparative Example 9

[0069] This comparative example provides a method for preparing an extract of Sparganii Rhizoma, and the difference from Example 1 is that: only two reflux extractions are carried out, and the other conditions are the same.

[0070] Effect Example 1

[0071] In this example of effects, the extracts of Sparganium stoloniferum prepared in Examples 1-3 and Comparative Examples 1-9 were used as samples to explore the contents of flavonoid and polyphenol active ingredients. The specific method is as follows:

[0072] Table 1

[0073]

[0074]

[0075] As can be seen from the results in Table 1, the Sparganium stoloniferum powder with smaller particles has a larger specific surface area and can be more effectively dissolved and extracted in the solvent, thus increasing the content of flavonoids and polyphenols. Appropriate particle size is more likely to remain stable during the extraction process, while overly fine particles may be more easily damaged under the action of the solvent, resulting in the degradation or oxidation of some flavonoids and polyphenols, thus affecting the final content. Therefore, it is most appropriate to select 40-60 mesh; the continuous high temperature and concentration gradient of heating under reflux are more conducive to the reaction, which can promote the release and dissolution of flavonoids and polyphenols. However, the ultrasonic extraction time is short and cannot achieve the effect of sufficient extraction. Therefore, the heating under reflux extraction method is selected as the best; if the volume fraction of ethanol is too low and the polarity is strong, it may not be able to effectively dissolve flavonoids and polyphenols, and at the same time, it cannot effectively break the plant cell wall, making it difficult for the flavonoids and polyphenols inside the cells to be released into the solvent. If the volume fraction of ethanol is too high, the dissolution amounts of alcohol-soluble and fat-soluble substances increase, resulting in a decrease in the dissolution rate of flavonoid compounds. Therefore, it is best to select 60-70% for the volume fraction of ethanol; when the temperature is too low, the penetration and dissolution of the solvent ethanol on Sparganium stoloniferum cells are weakened, and the extraction rate becomes slower, making the extraction of flavonoid and polyphenol active substances insufficient. When the temperature is too high, the dissolution amount of impurities increases, hindering the dissolution of flavonoid and polyphenol substances. Therefore, it is best to select 75°C - 85°C for the heating under reflux temperature; if the number of extractions is too small or the extraction time is too short, the effective active ingredients such as flavonoids and polyphenols in Sparganium stoloniferum cannot be completely extracted. If the number of extractions is too large or the extraction time is long, the alcohol-soluble substances increase, and some flavonoid and polyphenol substances may be oxidized, resulting in a decrease in the extraction rate. Therefore, it is best to select three extractions and the extraction time for the third extraction to be 0.5-1.5 h.

[0076] It is precisely by comprehensively preparing and extracting the Sparganium stoloniferum extract under the above specific process conditions that the present invention can maximize the extraction efficiency of flavonoid and polyphenol active ingredients in Sparganium stoloniferum, so as to achieve a better post-sun damage repair effect when applied to products such as cosmetics.

[0077] Example of effects 2

[0078] 1. Antioxidant test:

[0079] (1) DPPH free radical scavenging:

[0080] After diluting the triangular prism extract prepared in Example 1 to different concentrations, referring to the "Cosmetics - Free Radical (DPPH) Scavenging Experiment Method", 10 mL test tubes were used to set up sample tubes (T), sample backgrounds (T 0 ), DPPH tubes (C), and solvent backgrounds (C 0 ). For each test concentration of each sample, 3 parallel tubes were required for the sample tube (T), and 3 parallel tubes were also required for the DPPH tube (C). Add 1 mL of the triangular prism extract solution with the same concentration to the sample tube (T) and the sample background (T 0 ). Supplement the solvent in all test tubes (T, T 0 , C, C 0 ), use water for water-soluble samples and 95% ethanol for oil-soluble samples to make up to 3 mL, and mix well. Add 1 mL of DPPH ethanol solution to the sample tube (T) and the DPPH tube (C), and replace the sample background (T 0 ) and the solvent background (C 0 ) with 95% ethanol, shake gently, and let stand at room temperature for 5 minutes. Transfer each reaction solution into a 1 cm cuvette and measure the absorbance at 517 nm. Calculate the DPPH free radical scavenging rate:

[0081] Scavenging rate (%) = [1 - (T - T 0 ) / (C - C 0 )] × 100%;

[0082] In the formula:

[0083] T - Absorbance of the sample tube, that is, the absorbance of the solution after the sample reacts with DPPH;

[0084] T 0 - Absorbance of the sample background;

[0085] C - Average value of the absorbance of the DPPH tube three times, that is, the absorbance of the DPPH solution without adding the sample;

[0086] C 0 - Absorbance of the solvent background.

[0087] The experimental results are shown in Table 2. The triangular prism extract has a good scavenging effect on DPPH in the concentration range of 0.5 - 0.0625 mg / ml, and the higher the concentration, the higher the scavenging rate.

[0088] Table 2

[0089] Concentration (mg / ml) 0.0625 0.125 0.25 0.5 Clearance rate (%) 32.79 48.41 60.76 78.86 SD (%) 0.68 0.92 0.51 0.49

[0090] (2) Hydroxyl radical scavenging rate:

[0091] After diluting the triangular prism extract prepared in Example 1 with ethanol-water to different concentrations, referring to the "o-Phenanthroline Method for Scavenging Hydroxyl Radicals", 1 mL of o-phenanthroline solution, 1.5 mL of phosphate buffer solution, and 1 mL of deionized water were successively taken into a 25 mL volumetric flask, thoroughly mixed, 1 mL of ferrous sulfate solution was added, and after mixing, 1 mL of hydrogen peroxide was added, and the volume was made up with deionized water; it was placed in a water bath at 37 °C for 60 min, and the absorbance was measured at 510 nm, which was the absorbance A damage of the damaged flask; for the undamaged flask, 1 mL of deionized water was used to replace 1 mL of hydrogen peroxide in the damaged flask, and the absorbance A undamaged of the undamaged flask at 510 nm could be measured. The sample flask was replaced with 1 mL of the triangular prism extract solution instead of 1 mL of deionized water in the damaged flask. Calculate the hydroxyl radical scavenging rate:

[0092] CL=(A 样品 -A 损伤 ) / (A 未损 -A 损伤 )×100

[0093] In the formula: CL - the hydroxyl radical scavenging rate of the sample, %

[0094] A 样品 - the absorbance value of the sample;

[0095] A 未损 - the absorbance value of the undamaged group;

[0096] A 损伤 - the absorbance value of the damaged group.

[0097] The experimental results are shown in Table 3. The triangular prism extract has a good scavenging effect on hydroxyl radicals in the concentration range of 5 - 20 mg / ml, and the higher the concentration, the higher the scavenging rate.

[0098] Table 3

[0099] Concentration (mg / ml) 5 10 20 Clearance rate (%) 5.77 13.46 53.85 SD (%) 0.60 0.96 1.21

[0100] (3) Lipid peroxidation experiment:

[0101] Ferric thiocyanate method (FTC): The ferric thiocyanate (FTC) colorimetric method is based on the fact that under acidic conditions, the peroxide formed by lipid oxidation can oxidize Fe 2+ to Fe 3+ , and then Fe 3+A red complex that has a maximum absorption within 480 - 515 nm is formed with thiocyanate ions to measure the absorbance. The rhizoma sparganii extract prepared in Example 1 was formulated into a 20 mg / mL solution with methanol as the test sample solution; 2 mL of the above sample solution was taken, added with 2 mL of linoleic acid solution and 4 mL of PBS buffer, and the blank was measured with methanol solution; the control solution and the sample solution were placed in a constant temperature incubator at 40 °C for 24 h, 48 h, 72 h, 96 h, and 120 h and then measured.

[0102] Measurement method: Take 0.1 mL of the sample solution, add 4.7 mL of 75% ethanol solution, 0.1 mL of 30% ferric thiocyanate solution, and 0.1 mL of ferrous chloride solution, and mix well; let stand for 3 min, and measure the absorbance value of the mixture at a wavelength of 500 nm. Calculate the inhibition rate of linoleic acid peroxidation:

[0103] Inhibition rate of linoleic acid peroxidation = [100 - (OD 样品 / OD 对照 )] × 100%

[0104] The experimental results are shown in Table 4. The rhizoma sparganii extract has a good inhibitory effect on linoleic acid peroxidation at a concentration of 10 mg / ml.

[0105] Table 4 Inhibition rate of linoleic acid peroxidation of 10 mg / ml rhizoma sparganii extract

[0106]

[0107]

[0108] (4) ROS scavenging experiment:

[0109] Expose 24 48-hpf zebrafish embryos to the 0.5% rhizoma sparganii extract solution prepared in Example 1. At the same time, set up a blank control group and a positive control group (0.5% glutathione). After 24 h of exposure, the fish embryos were stained with H2DCFDA, and fluorescence photography was used to measure the ROS signal intensity and perform statistical analysis. Calculate the reactive oxygen species (ROS) scavenging rate:

[0110] Scavenging rate = (B - S) / B × 100%;

[0111] In the formula:

[0112] S—the average value of the "average signal intensity" of ROS in the fish embryos of the test substance treatment group;

[0113] B—the average value of the "average signal intensity" of ROS in the fish embryos of the blank control group;

[0114] The results are as Figure 1As shown, under the same test conditions, the ROS inhibition rate of glutathione is 15%, while the extract of Sparganium stoloniferum prepared by the present invention has a ROS scavenging rate of 24%, showing good antioxidant efficacy.

[0115] (5) AGEs Scavenging Experiment:

[0116] The extract of Sparganium stoloniferum prepared in Example 1 was formulated into a 1% solution with deionized water for testing. A mixed solution containing BSA and glucose was prepared with phosphate buffer (PBS) and filtered as a 2× glycation reaction solution. Reaction systems of each group were prepared in test tubes A (1 ml of 2× glycation reaction solution, 1 ml of sample / reference solution), B (1 ml of sample / reference solution, 1 ml of PBS), C (1 ml of 2× glycation reaction solution, 1 ml of sample / reference solution), and D (1 ml of sample / reference solution, 1 ml of PBS). After mixing evenly, they were placed in a digital display constant temperature water bath for incubation for 4 days. Phosphate buffer (PBS) was used as a negative control instead of the sample, aminoguanidine hydrochloride solution was used as a positive control, and phosphate buffer (PBS) was used as a control system instead of the glycation reaction solution. After the reaction, the incubated solution was cooled to room temperature for determination. 200 μL of the reaction solution was taken and added to a 96-well plate in sequence, and the fluorescence intensity was detected using a multifunctional microplate reader under the conditions of an excitation wavelength of 370 nm and an emission wavelength of 440 nm, and the AGEs inhibition rate was calculated.

[0117] AGEs inhibition rate (%) = [(C - D) Mean - (A - B)] / (C - D) Mean × 100%;

[0118] In the formula:

[0119] A: Fluorescence intensity of the glycation system with the test substance added;

[0120] B: Fluorescence intensity of the PBS solution with the test substance added;

[0121] C: Fluorescence intensity of the glycation system without the test substance added;

[0122] D: Fluorescence intensity of the PBS solution without the test substance added.

[0123] The experimental results are shown in Table 5. The AGEs inhibition rate of the Sparganium stoloniferum extract at a concentration of 1% is 19.10%, which is significantly higher than the AGEs inhibition rate of the negative control (P < 0.05), indicating that the Sparganium stoloniferum extract has antiglycation efficacy.

[0124] Table 5 AGEs Inhibition Results of 1% Sparganium stoloniferum Extract

[0125] Group AGEs Inhibition rate (average value) Standard deviation Significant difference Sample group 19.10％ 3.52％ P＜0.05 Positive control group 91.04％ 2.74％ P＜0.05 Negative control group 0.00％ 11.80％ /

[0126] Whether oxidative stress can be effectively inhibited is one of the important indicators for evaluating the therapeutic effect of products on post-sun damage. Therefore, in summary, the rhizoma sparganii extract prepared by the present invention can effectively scavenge free radicals, inhibit lipid peroxidation and the expression of ROS, reduce the generation of AGEs, and reduce oxidative stress response, thereby reducing post-sun skin damage caused by ultraviolet rays.

[0127] 2. Inflammatory response inhibition test:

[0128] By irradiating keratinocytes with UVB, the gene expressions of NF-κB and HMGB1 and the content of TNF-α were measured.

[0129] (1) Cytotoxicity test:

[0130] The MTT method was used to detect cytotoxicity and determine the administration concentration of the rhizoma sparganii extract on keratinocytes. After resuscitating the cells, when the plating rate reached about 60%, the cells were seeded into 96-well plates and incubated overnight in a CO 2 incubator (37 °C, 5% CO 2 ). The test was set up with a zero control group, a solvent control group, a positive control group and a sample group. The sample group was the rhizoma sparganii extract, and 8 concentrations (0.5%, 0.25%, 0.125%, 0.0625%, 0.0313%, 0.0156%, 0.0078%, 0.0039%) were set, and 3 replicate wells were set for each concentration. When the plating rate of the cells in the 96-well plates reached 50%-60%, the drug was administered. The solvent control group added 200 μL of culture medium to each well; the positive control group added 200 μL of culture medium containing 10% DMSO to each well; the sample group added 200 μL of culture medium containing the corresponding concentration of the sample to each well; the zero control group had no cell seeding and only added 200 μL of cell culture medium. After the drug administration was completed, the 96-well plates were placed in a CO 2 incubator (37 °C, 5% CO 2 ) and cultured for 24 h. After the cells were incubated for 24 h, the supernatant was discarded, MTT working solution (0.5 mg / mL) was added, and incubated at 37 °C in the dark for 4 h. After the incubation was completed, the supernatant was discarded, 150 μL of DMSO was added to each well, and the OD value was read at 490 nm. Calculated according to the formula:

[0131] Relative cell viability (%) = (OD of sample well - OD of zero control well) / (OD of solvent control well - OD of zero control well) × 100%;

[0132] The test results are shown in Table 6. The rhizoma sparganii extract did not show obvious cytotoxicity based on keratinocytes within the concentration range of 0.0156%.

[0133] Table 6 MTT test results of the rhizoma sparganii extract on keratinocytes

[0134]

[0135] (2) Detection of the inhibition of NF-κB, HMGB1, and TNF-α expression:

[0136] After resuscitating the cells, when the plating rate reaches about 60%, inoculate the cells into a 6-well plate and place them in a CO 2 incubator (37 °C, 5% CO 2 ) and incubate overnight. When the plating rate of the cells in the 6-well plate reaches 30%-50%, perform grouping and drug administration, with 3 replicate wells in each group. Add 2 mL of culture medium to each well in the blank control group and the negative control group, add 2 mL of culture medium containing dexamethasone to each well in the PC group, and add 2 mL of culture medium containing 0.02% Sparganium stoloniferum extract to each well in the sample group. Except for the blank control group, the other groups are irradiated with UVB at an irradiation dose of 300 mJ / cm 2 , and after the irradiation is completed, place them in a CO 2 incubator (37 °C, 5% CO 2 ) and culture for 24 h. After the incubation is completed, collect the cell culture supernatant, and perform ELISA to detect TNF-α according to the instructions of the ELISA kit. Wash the cells twice with PBS, add 1 mL of RNAisoPlus to each well, blow and lyse the cells, and then collect the samples. Extract the RNA, reverse transcribe it into cDNA, and then perform fluorescence quantitative PCR to detect the expression results of the HMGB1 and NF-κB genes. Use the 2 -△△CT method to calculate the results.

[0137] Inhibition rate (%) = (negative control group - sample group) / negative control group × 100%;

[0138] The results are shown in Tables 7-9. The 0.02% Sparganium stoloniferum extract effectively inhibits the gene expression levels of NF-κB and HMGB1 and the content of TNF-α, and the inhibition rates are 59.31%, 33.57%, and 20.00% respectively.

[0139] When performing statistical analysis using the t-test method, compared with the BC group, the significance is indicated by #, P-value < 0.05 is indicated by #, and P-value < 0.01 is indicated by ##; compared with the NC group, the significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0140] Table 7 Summary of the detection results of the NF-κB gene

[0141]

[0142] Table 8 Summary of the detection results of the HMGB1 gene

[0143]

[0144] Summary of TNF-α content detection results in Table 9

[0145]

[0146] The inflammatory signaling pathway protein NF-κB is a nuclear transcription factor. When phosphorylated and activated, it enters the nucleus and binds to the promoter region of downstream target genes, initiating the transcription of downstream target genes. The downstream target genes are mainly factors related to the inflammatory response. Therefore, inhibiting NF-κB can inhibit the inflammatory response. HMGB1 functions to recruit inflammatory cells and mediate signals among macrophages, dendritic cells, and natural killer cells. Extracellular HMGB1 can activate endothelial cells, promote angiogenesis, enhance the migration of hematopoietic stem cells, and trigger local or systemic inflammation. TNF-α is a pro-inflammatory factor, the earliest and most important inflammatory mediator in the inflammatory response process. It can activate neutrophils and lymphocytes, increase the permeability of vascular endothelial cells, regulate the metabolic activity of other tissues, and promote the synthesis and release of other cytokines, inducing the occurrence of the inflammatory response.

[0147] In summary, based on keratinocytes, compared with the control group, the extract of Sparganium stoloniferum can significantly down-regulate the expression of NF-κB and HMGB1 genes induced by ultraviolet rays, inhibit the secretion of inflammatory factors (TNF-α), reduce the inflammatory response, and thus reduce the post-sunburn damage of the skin caused by ultraviolet rays.

[0148] 3. Pigmentation inhibition test:

[0149] (1) Irradiate keratinocytes with UVB and measure the content of α-MSH and the gene expression of β-catenin

[0150] Same as the inflammatory response inhibition test, after cytotoxicity MTT testing, the extract of Sparganium stoloniferum with a concentration of 0.02% was finally selected for subsequent experiments. After resuscitating the cells, when the plating rate reached about 60%, the cells were seeded into 6-well plates and incubated overnight in a CO 2 incubator (37 °C, 5% CO 2 ). When the plating rate of the cells in the 6-well plates reached 30%-50%, grouping and drug administration were carried out, with 3 replicate wells in each group. 2 mL of culture medium was added to each well in the blank control group and the negative control group, 2 mL of culture medium containing VE was added to each well in the PC group, and 2 mL of culture medium containing 0.02% extract of Sparganium stoloniferum was added to each well in the sample group. Except for the blank control group, the other groups were irradiated with UVB at an irradiation dose of 300 mJ / cm 2 , after the irradiation, it was placed in a CO 2 incubator (37 °C, 5% CO 2) Incubate for 24 h. After the incubation, collect the cell culture supernatant and perform ELISA to detect α-MSH according to the ELISA kit instructions. Wash the cells twice with PBS, add 1 mL of RNAisoPlus to each well, pipette to lyse the cells, and collect the samples. Extract RNA, reverse transcribe it into cDNA, and then perform fluorescence quantitative PCR to detect the expression results of the β-catenin gene. Use the 2 -△△CT method for result calculation.

[0151] Inhibition rate (%) = (negative control group - sample group) / negative control group × 100%;

[0152] The results are shown in Table 10 - 11. The 0.02% extract of Sparganii Rhizoma effectively inhibits the content of α-MSH and the gene expression level of β-catenin, with inhibition rates of 53.34% and 54.23% respectively.

[0153] When performing statistical analysis using the t-test method, compared with the BC group, significance is indicated by #, P-value < 0.05 is indicated by #, and P-value < 0.01 is indicated by ##; compared with the NC group, significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0154] Table 10 Summary of the detection results of α-MSH content

[0155]

[0156] Table 11 Summary of the detection results of the β-catenin gene

[0157]

[0158] α-MSH is mainly produced through autocrine and paracrine mechanisms by melanocytes and keratinocytes, and keratinocytes are the main source of α-MSH. α-MSH can rapidly increase the activity of tyrosinase, stimulate the increase of melanocyte dendrites, and promote the transfer of melanosomes to surrounding keratinocytes. β-catenin accumulates in the cytoplasm and then translocates to the nucleus, which is considered a marker of the activation of the Wnt signaling pathway. The secreted glycoprotein Wnt plays an important role in the development of melanocytes. In the Wnt / β-catenin signaling pathway, after the Wnt ligand binds to the membrane receptor G protein-coupled receptor Frizzled, β-catenin is not phosphorylated and degraded by blocking the activity of glycogen synthase kinase. The accumulated β-catenin is transported to the nucleus through Rac1 and other factors and binds to the lymphoid enhancer factor-1 / T cell factor (LEF-TCF) transcription complex, resulting in the promotion of MITF expression by the increased concentration of β-catenin in the nucleus, thereby promoting the synthesis of intracellular melanin.

[0159] In summary, 0.02% of the extract of Sparganium stoloniferum, based on keratinocytes, significantly downregulates the content of α-melanocyte-stimulating hormone (α-MSH) and the gene expression level of β-catenin compared with the control group, and effectively inhibits signal transduction during ultraviolet-induced melanogenesis.

[0160] (2) Inhibiting melanogenesis:

[0161] Similarly, the MTT method was used to determine the cytotoxicity of the extract of Sparganium stoloniferum in melanocytes, and finally, 0.03% concentration of the extract of Sparganium stoloniferum was selected for the melanin inhibition test. The cytotoxicity results are shown in Table 12 below:

[0162] Table 12 MTT test results of the extract of Sparganium stoloniferum on B16 cells

[0163]

[0164] Refer to "In Vitro Test of B16 Cell Melanin Synthesis Inhibition Experiment for Cosmetics" (T / SHRH027-2019) to determine the effect of the extract of Sparganium stoloniferum on the melanin content of cells: adjust the cell density to 2.5×10 3 cells / mL with cell culture medium, and then spread the cell suspension in a 6-well plate, 2 mL per well. Note: Each test needs to set a negative control group, a test sample group, and a positive control group, and three replicates are set for each group. The prepared 6-well plate is placed at 37°C, 5% CO 2After being placed in the incubator for 20 - 24 hours, the cell culture medium was replaced with the culture medium containing the test sample, 2 mL per well, and continued to be cultured for 48 hours. 48 hours after the first medium change, repeat the steps, replace the cell culture medium, adjust to add 3 mL of the culture medium containing the corresponding sample per well, and continue to culture in a 37°C, 5% CO 2 incubator. 72 hours after the second medium change, repeat the steps, replace the cell culture medium, add 3 mL of the culture medium containing the sample per well, and continue to culture in a 37°C, 5% CO 2 incubator. 48 - 72 hours after the third medium change (when the cell fusion rate reaches over 90%), measure the melanin content of the cells. Wash the cells twice with 1×PBS, aspirate the PBS, add 200 μL of 0.25% trypsin to each well to digest the cells, and place them in a CO 2 incubator for digestion for 5 minutes. Add 2 mL of PBS to blow and beat the cells in two portions, collect the cells into a centrifuge tube, and centrifuge at 4000 rpm for 5 minutes. Try to aspirate as much of the remaining PBS in the centrifuge tube as possible, add 200 μL of melanin extraction solution to each tube, shake well, and place in an 80°C water bath for heating for 1 hour. After cooling, centrifuge the droplets on the lower wall of the tube moderately, blow and beat evenly, aspirate 150 μL of the solution from each centrifuge tube, transfer it to a 96-well plate, and use an enzyme-linked immunosorbent assay (ELISA) reader to detect the absorbance at 405 nm of each well. Calculate the melanin synthesis inhibition rate:

[0165] Inhibition rate (%) = (C - T) / (C - C 0 ) × 100%;

[0166] In the formula:

[0167] T - Absorbance of the test sample well;

[0168] C - Three-time average value of the absorbance of the negative control group;

[0169] C 0 - Background absorbance of the melanin extraction solution.

[0170] The results are shown in Table 13. The melanin inhibition rate of the 0.03% extract of Sparganium stoloniferum is 33.08%. The generation of melanin by melanocytes in the skin is the main cause of skin darkening and the formation of skin spots, indicating that the extract of Sparganium stoloniferum can effectively inhibit the generation of melanin.

[0171] Table 13 Inhibitory results of the extract of Sparganium stoloniferum on the melanin content in B16 cells

[0172]

[0173] (3) Co-culture of keratinocytes and melanocytes to measure the transport of melanosomes

[0174] Similarly, the MTT method was used to determine the cytotoxicity of the extract of Sparganii Rhizoma in the co-culture system of keratinocytes and melanocytes. Finally, extracts of Sparganii Rhizoma at concentrations of 0.12% and 0.06% were selected for the melanosome transport test experiment. The cytotoxicity results are shown in Table 14 below:

[0175] Table 14 MTT detection results of the co-culture of keratinocytes and melanocytes with the extract of Sparganii Rhizoma

[0176]

[0177] Cells in the logarithmic growth phase were digested with 0.25% trypsin and centrifuged, and then prepared into a cell suspension with an appropriate density using complete medium. HaCaT cells and A375 cells were prepared into a cell suspension at a ratio of 10:1; a cell slide was placed in each well of a 12-well plate, and 1 mL was inoculated into each well of the 12-well culture plate according to the experimental requirements and cultured in a 37°C, 5% CO 2 incubator for 24 ± 2 h. The original culture medium in the 12-well plate was removed, and sample addition treatments were performed according to the experimental groups. In the experimental groups, a culture medium containing extracts of Sparganii Rhizoma at concentrations of 0.12% and 0.06% and an α-MSH inducer were added to each well. In the model group, the same concentration of α-MSH inducer was added, and in the blank control group, only the culture medium was added, 1 mL per well, and the cells were further cultured in a 37°C, 5% CO 2 incubator for 24 ± 2 h. After incubation, the original culture medium was discarded, and the cells were washed 2 - 3 times with PBS. Fixative was added to each well and fixed for 20 min, and then blocked. The primary antibody was added and incubated at 2 - 8°C overnight or incubated in a 37°C incubator for 1.5 h; the cells were washed 2 - 3 times with PBS, the secondary antibody was added and incubated at room temperature for 1 h or incubated in a 37°C incubator for 40 min, and finally, the nuclei were stained with Hoechst33342 live cell staining solution for 10 min. Note: From the addition of the secondary antibody, the operation was carried out in a dark room throughout to avoid fluorescence quenching. The cell slides were transferred to a dark room and photographed with a fluorescence microscope, and the fluorescence intensity of the collected photos was analyzed using ImageJ software.

[0178] Calculate the absolute fluorescence intensity of melanosomes:

[0179] Absolute fluorescence intensity of melanosomes = average fluorescence intensity of melanosomes / average fluorescence intensity of cell nuclei. Formula for calculating the relative content of melanosomes:

[0180] Relative content of melanosomes (%) = T / C × 100%;

[0181] Where:

[0182] T—the absolute fluorescence intensity value of melanosomes in the test sample;

[0183] C—the average absolute fluorescence intensity of melanosomes in the model group.

[0184] Calculation formula for the melanosome transport inhibition rate:

[0185] Inhibition rate (%) = (1 - T / C) × 100%;

[0186] In the formula:

[0187] T — Average value of the relative content of melanosomes in the test sample;

[0188] C — Average value of the relative content of melanosomes in the model group.

[0189] The results are as Figure 2 - 3 shown: The results of fluorescence microscopy observation (red fluorescence indicates melanosomes, and the brighter the fluorescence, the more melanosomes) show that when the extracts of Sparganium stoloniferum at concentrations of 0.12% and 0.06% act on the co - culture system of HaCaT cells and A375 cells, the relative contents of melanosomes are 7.45% and 43.84% respectively. Compared with the model group (100%), the relative contents of melanosomes are significantly reduced (P < 0.05). The melanosome transport inhibition rates are 92.55% and 56.16% respectively.

[0190] When irradiated with ultraviolet light, the melanin precursors in cells are oxidized, and melanin synthesis and redistribution occur. Keratinocytes cooperate in transporting and accumulating it, ultimately changing the skin pigmentation. Melanin is produced in special dendritic cells, namely melanocytes. Melanin is synthesized through a cascade reaction triggered by tyrosinase, in which tyrosine is converted into melanin. During the continuous deposition of melanin in melanosomes, it is simultaneously transported to the dendritic ends of melanocytes. Melanosomes will be transferred to keratinocytes through possible mechanisms such as endocytosis - exocytosis and membrane fusion, thus presenting skin darkening.

[0191] In summary, the extracts of Sparganium stoloniferum can not only effectively inhibit the generation of new melanin but also inhibit the transport of the already - generated melanin, and have a significant improvement effect on ultraviolet - induced skin pigmentation.

[0192] Effect Example 3

[0193] The extract of Sparganium stoloniferum prepared in Example 1 was formulated into an emulsion dosage form for subsequent clinical human tests, and the content of the extract of Sparganium stoloniferum was 2%.

[0194] 1. The formula and process are as follows:

[0195] Phase A: Glycerol 3.0 wt%, Polyacrylate crosspolymer - 6 0.35 wt%, Sodium metabisulfite 0.35 wt%, Xanthan gum 0.06 wt%, Sodium hyaluronate 0.06 wt%, p - Hydroxyacetophenone 0.50 wt%, 1,2 - Hexanediol 0.60 wt%, Propylene glycol 4.00 wt%;

[0196] Phase B: 2.00 wt% squalane, 3.00 wt% glycerin caprylate / caprate, 4.00 wt% isopropyl myristate, 0.15 wt% tocopherol, 0.60 wt% MONTANOV TM L, 0.30 wt% stearic acid, 2.00 wt% cetearyl alcohol, 1000 1.50 wt%, 0.40 wt% span 60;

[0197] Phase C: 2.00 wt% extract of Rhizoma Sparganii, the balance being deionized water.

[0198] The preparation method of the emulsion comprises the following steps:

[0199] (1) Accurately weigh Phase A, heat it in a water bath at 80°C, homogenize it rapidly for 5 min until uniform, and set aside;

[0200] (2) Accurately weigh Phase B, heat and dissolve Phase B, add Phase B to Phase A, homogenize for 5 min, and then stir;

[0201] (3) When the temperature of the material drops below 50°C, add Phase C, stir it evenly, cool it to room temperature, and discharge it to obtain the emulsion containing the extract of Rhizoma Sparganii, which is used for subsequent human trials.

[0202] 2. Safety test:

[0203] (1) Subjects: 30 outdoor workers (20 females and 10 males) were screened out, showing characteristics such as facial skin flushing, damaged skin barrier, uneven skin tone, obvious pigmentation, etc. Their ages were between 25 and 65 years old. All subjects had no history of skin or systemic diseases, the test sites were normal, and no drugs or cosmetics unrelated to the experiment were applied during the test period.

[0204] (2) Test environment: The test site was kept at a constant temperature and humidity, the environmental temperature was 20°C - 22°C, and the relative humidity was 40% - 60%. The subjects should keep their bodies in a stable state before the test. After washing the subjects' faces with water at about 35°C, they should sit still in the test environment for 30 min before starting the test.

[0205] (3) Test method: The emulsion containing the extract of Rhizoma Sparganii prepared above was used for patch testing. Qualified patch test devices were selected. By the closed patch test method, about 0.020 - 0.025 ml of the test substance was placed in the patch test device, and the patch test device was applied externally to the flexor side of the forearm of the subject with medical tape. It was gently pressed with the palm to make it evenly applied to the skin. After 24 hours, the patch test device of the test substance was removed, and the skin reactions were observed at 0.5, 24, and 48 hours after removing the patch test device of the test substance.

[0206] The results of the human skin patch test showed that among 30 people, there were no skin adverse reactions, and the sample was safe and non-irritating.

[0207] 3. Human testing:

[0208] The above-mentioned 30 selected subjects were subjected to human trials to explore the effect of the prepared lotion containing Rhizoma Sparganii extract on improving skin damage after sun exposure.

[0209] Method of using the sample: Twice a day, once after morning and evening face cleansing, about 1 g of the sample each time, and continuously used for 56 days.

[0210] The cheeks of the selected volunteers were used as the test sites, and VISIA tests were carried out before using the test product and on the 28th and 56th days of continuous use of the product. The comparison of the pigmented spots, ultraviolet pigmented spots, brown spots and red areas of a representative subject before use and after 56 days of use is as Figure 4 - 7 shown. Among them, the pigmented spot feature count decreased by 3.3%, the ultraviolet pigmented spot feature count decreased by 2.7%, the brown spot feature count decreased by 16.6%, and the red area feature count decreased by 22.9%. This shows that the lotion containing Rhizoma Sparganii extract can effectively reduce the skin inflammatory reaction and sunburn reaction caused by ultraviolet rays, reduce flushing, as well as pigmentation such as pigmented spots, ultraviolet pigmented spots and brown spots, and play an obvious effect in improving skin damage after sun exposure.

[0211] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. The use of the extract of Tripterygium wilfordii in the preparation of an external preparation for skin after-sun repair, characterized in that: The preparation method of the Tripterygium wilfordii extract comprises the following steps: (1) Crush the tuber of Tripterygium wilfordii, pass it through a 40-60 mesh sieve, soak it in an ethanol solution, reflux extract it for 1.5-2.5 hours, filter it to obtain a first extract, and then reflux extract the filter residue with an ethanol solution for 1.5-2.5 hours, filter it, and obtain a second extract; (2) refluxing the filter residue with an ethanol solution for 0.5-1.5 hours to obtain a third extract, combining the three extracts, removing the ethanol under reduced pressure, filtering and collecting the filtrate to obtain the Tripterygium wilfordii extract; The temperatures of the three reflux extractions are all 75-85°C; The volume concentration of the ethanol solution is 60-70%.

2. The use according to claim 1, characterized in that The extract of Tripterygium wilfordii exerts the effect of repairing skin after sun exposure through at least one of the following aspects: a. Inhibit oxidative stress caused by sun damage to skin tissue; b. Inhibit the inflammatory response caused by sun damage to skin tissue; c. Inhibit pigmentation caused by sun damage to skin tissue.

3. A cosmetic for skin repair after sun exposure, characterized in that: The cosmetic comprises the Tripterygium wilfordii extract used in the application of claim 1 or 2, and the mass percentage of the Tripterygium wilfordii extract in the cosmetic is 1-60%.

4. The cosmetic according to claim 3, characterized in that The cosmetic dosage form includes at least one of an aqueous solution, an emulsion, and a cream.

5. A skin after-sun repair lotion, characterized in that: The emulsion comprises the Tripterygium wilfordii extract used in the application of claim 1 or 2; the mass percentage of the Tripterygium wilfordii extract in the emulsion is 1-3%.

6. The emulsion according to claim 5, characterized in that The emulsion further comprises 10-15% of a moisturizing agent, 5-10% of an emulsifier, 0.2-0.7% of an antioxidant, 1-3% of a thickener, 1-2% of a preservative and the balance of water.

7. The emulsion according to claim 6, characterized in that The moisturizing agent includes at least one of glycerin, sodium hyaluronate, propylene glycol, squalane, caprylic / capric glyceride, glucose, butylene glycol, 1,2-pentanediol, caprylyl glycol and isopentyl glycol; The emulsifier includes at least one of acrylates / acrylamide copolymer, isopropyl myristate, C14-22 alcohol, C12-20 alkyl glucoside, stearic acid, cetearyl olivate, sorbitan olivate, Span 80, glyceryl stearate, PEG-100 stearate, steareth-2, steareth-21, ceteareth-12, ceteareth-30, potassium cetyl phosphate, sodium stearoyl glutamate, polysorbate-60 and sorbitan stearate; The antioxidant comprises at least one of tocopherol and tocopherol acetate; The thickener comprises at least one of polyacrylate crosspolymer-6, xanthan gum, cetearyl alcohol, carbomer and hydroxyethyl cellulose; The preservative comprises at least one of p-hydroxyacetophenone, 1,2-hexanediol, caprylhydroxamic acid, benzyl alcohol, phenoxyethanol, benzoic acid and its salts, and sorbic acid and its salts.

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