Medical composition for repairing and relieving after art as well as preparation method and application thereof

Abstract

The invention discloses a composition for post-art repairing and relieving as well as a preparation method and application thereof, and relates to the technical field of cosmetics. The invention provides a composition for repairing and relieving. The composition is prepared from the following components in parts by weight: 0.1 to 1 part of citrus fruit extract, 0.1 to 0.5 part of phoenix fruit extract, 0.05 to 0.5 part of radix gentianae extract and 0.1 to 1 part of lipidosome, the liposome is prepared from dipotassium glycyrrhizinate and phytol. The composition for repairing and relieving provided by the invention realizes rapid repairing and long-acting stability maintenance through a multi-dimensional synergistic mechanism. The composition disclosed by the invention can be used for dynamically regulating the pH value of the skin in a synergetic manner, immediately blocking neurogenic pain signal conduction mediated by a TRPV1 receptor, synergistically reconstructing a physical barrier and a micro-ecological barrier, promoting vasoconstriction and activating a skin TRPM skin cold feeling body so as to achieve the effect of'cold compress'.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a composition for repairing and soothing after medical aesthetics, a preparation method thereof, and an application thereof. Background Art

[0002] In the current field of medical aesthetics, photoelectric therapies (including intense pulsed light, laser, radiofrequency, etc.) and injection techniques (such as botulinum toxin, hyaluronic acid filling) have become conventional treatment means for improving problems such as skin relaxation, skin pigmentation, and acne. Although such techniques can achieve a "break and then stand" regeneration and repair mechanism through controllable skin damage, there are still significant postoperative complications in clinical applications: ① The thermal damage caused by photoelectric treatment can destroy the continuous structure of the stratum corneum, leading to an abnormal increase in the transcutaneous water loss rate (TWEL), resulting in persistent skin dryness, sensitivity, and erythema reaction; ② Injection operations are prone to cause mechanical damage to vascular endothelium, induce local tissue ischemic lesions, form secondary damages such as vascular embolism and subcutaneous induration, and at the same time, the puncture channel creates an invasion path for the colonization of pathogenic microorganisms.

[0003] In response to the above problems, existing repair techniques mainly use physical barrier repair agents such as hyaluronic acid moisturizing gel and collagen dressing, and are supplemented with anti-inflammatory components for symptom control. However, through experimental verification, this technical system has the following key defects: (1) Lack of acid-base balance regulation mechanism: The exudate from the postoperative wound surface causes abnormal fluctuations in the skin surface pH value, directly affecting the endogenous biosynthesis pathway of ceramide and delaying the barrier repair process; (2) Insufficient pain regulation efficacy: Existing components lack the ability to immediately block the TRPV1 receptor-mediated neurogenic pain signal transduction; (3) Lack of reconstruction of the microbial barrier: The synergistic regulation mechanism with symbiotic flora such as Staphylococcus epidermidis and Propionibacterium acnes has not been established, increasing the risk of colonization by pathogenic bacteria such as Staphylococcus aureus and Pseudomonas aeruginosa after surgery; (4) Backward cryotherapy technology: The current ice pack cold compress has problems such as uncontrollable temperature gradient and inaccurate action site.

[0004] Based on this, a composition for repairing and soothing after medical aesthetics that achieves rapid repair and long-term stability through a multi-dimensional synergistic mechanism has broad application prospects. Summary of the Invention

[0005] Based on this, the object of the present invention is to overcome the above-mentioned deficiencies of the prior art and provide a composition for repairing and soothing after medical aesthetics, its preparation method and application. The composition for repairing and soothing provided by the present invention can instantaneously block the conduction of neurogenic pain signals mediated by TRPV1 receptors, promote vasoconstriction and activate the skin TRPM skin cold receptor, achieving the effect of "cold compress"; at the same time, it can dynamically adjust the skin pH, synergistically rebuild the physical barrier and the microecological barrier, realizing rapid repair and long-term stability. The composition for repairing and soothing after medical aesthetics provided by the present invention can achieve rapid repair and long-term stability through a multi-dimensional synergistic mechanism.

[0006] To achieve the above object, the technical solution adopted by the present invention is: a composition for repairing and soothing, comprising the following components in parts by weight: citrus fruit extract 0.1 - 1 part, date fruit extract 0.1 - 0.5 part, gentian root extract 0.05 - 0.5 part, liposome 0.1 - 1 part; the liposome comprises dipotassium glycyrrhizinate and phytol.

[0007] The composition for repairing and soothing provided by the present invention achieves rapid repair and long-term stability through a multi-dimensional synergistic mechanism, which is specifically embodied as follows: rapid repair is reflected in that the composition for repairing and soothing after medical aesthetics provided by the present invention can relieve discomfort such as postoperative pain, burning sensation, and redness and swelling, and long-term stability is reflected in that the composition for repairing and soothing after medical aesthetics provided by the present invention can rebuild the physical barrier and the microecological barrier, and maintain skin homeostasis for a long time, covering the entire postoperative repair cycle. The above composition of the present invention is synergistic, can instantaneously block the conduction of neurogenic pain signals mediated by TRPV1 receptors, promote vasoconstriction and activate the skin TRPM skin cold receptor, achieving the effect of "cold compress"; at the same time, it can dynamically adjust the skin pH, synergistically rebuild the physical barrier and the microecological barrier, realizing rapid repair and long-term stability. The inventors found that the liposome of the present invention comprises dipotassium glycyrrhizinate and phytol. As a water-soluble component, dipotassium glycyrrhizinate is encapsulated by the hydrophilic core of the liposome, and phytol, as a fat-soluble component, is embedded in the lipid bilayer to form a "hydrophilic-hydrophobic" biphasic carrier. The specific liposome structure of the present invention solves the problem of low solubility of phytol in water, and at the same time protects dipotassium glycyrrhizinate and phytol from photo-thermal degradation. The specific dipotassium glycyrrhizinate-phytol liposome of the present invention can better achieve rapid repair and long-term stability.

[0008] Preferably, the composition for repairing and soothing comprises the following components in parts by weight: citrus fruit extract 0.3 - 0.8 part, date fruit extract 0.2 - 0.3 part, gentian root extract 0.1 - 0.3 part, liposome 0.3 - 0.5 part.

[0009] During the actual experimental process, the inventors found that when the components in the above composition are within a specific weight range, rapid repair and long-term stability can be better achieved. The composition for post-aesthetic repair and soothing provided by the present invention can immediately block the TRPV1 receptor-mediated neurogenic pain signal transduction, promote vasoconstriction and activate the skin TRPM skin cold receptor, achieving the effect of "cold compress"; at the same time, it can dynamically regulate the skin pH, synergistically rebuild the physical barrier and the microecological barrier, and achieve rapid repair and long-term stability.

[0010] Preferably, based on the total weight of the composition for repair and soothing, the weight percentage of the liposome is 30-40%.

[0011] During the actual experimental process, the inventors found that when the weight percentage of the liposome is within the above range, it can better regulate the skin pH, synergistically rebuild the physical barrier and the microecological barrier, and better achieve long-term stability.

[0012] Preferably, the weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract is citrus fruit extract: date fruit extract: gentian root extract = (2-3):1:1.

[0013] During the actual experimental process, the inventors found that when the weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract is in the above specific ratio, the composition for repair and soothing has a better effect in relieving postoperative pain, burning sensation, redness and swelling, etc.

[0014] The preparation method of the liposome in the present invention can be one of the thin film dispersion method and the reverse evaporation method.

[0015] Preferably, the preparation method of the liposome includes the following steps:

[0016] (1) Mix soy lecithin, cholesterol, phytol, and 1,2-dioleoyl-3-trimethylammonium chloride alanine (DOTAP), and dissolve them in solution A, and then obtain a lipid film by rotary evaporation; solution A is a mixed solution of chloroform and ethanol;

[0017] (2) Add solution B to the lipid film in step (1), and perform ultrasonic hydration and homogenization to obtain the liposome; solution B is a phosphate buffer solution containing dipotassium glycyrrhizinate.

[0018] The present invention provides a method for preparing liposomes by the thin film dispersion method. When the liposomes prepared by the above method are combined with the citrus fruit extract, date fruit extract, and gentian root extract, rapid repair and long-term stability can be better achieved. And during the actual experimental process, the inventors found that the addition of DOTAP cationic lipid can neutralize the negative ions in dipotassium glycyrrhizinate, making the liposomes more stable.

[0019] Preferably, the weight ratio of soy lecithin, cholesterol, phytol, and 1,2-dioleoyl-3-trimethylammonium chloride alanine is soy lecithin: cholesterol: phytol: 1,2-dioleoyl-3-trimethylammonium chloride alanine = (50 - 70):(20 - 40):(5 - 10):(5 - 10);

[0020] and / or, the weight ratio of chloroform and ethanol is (2 - 3):1;

[0021] and / or, the mass-volume ratio of the mixture of soy lecithin, cholesterol, phytol, and 1,2-dioleoyl-3-trimethylammonium chloride alanine and solution A is (80 - 120) mg: 4 mL;

[0022] and / or, the mass concentration of solution B is 1 - 5 mg / mL; the pH of the phosphate buffer solution is 5.8 - 6.2;

[0023] and / or, the weight ratio of phytol and dipotassium glycyrrhizinate is 5:(4 - 25).

[0024] Preferably, the weight ratio of phytol and dipotassium glycyrrhizinate is 5:(8 - 20); more preferably, the weight ratio of phytol and dipotassium glycyrrhizinate is 5:(8 - 12).

[0025] Preferably, the temperature of rotary evaporation is 45 - 60 °C, the rotation speed of rotary evaporation is 100 - 200 rpm, and the time of rotary evaporation is 30 - 50 min;

[0026] and / or, the temperature of ultrasonic hydration is 50 - 60 °C, the time of ultrasonic hydration is 10 - 30 min, and the power of ultrasonic hydration is 100 - 200 w;

[0027] and / or, the pressure of homogenization is 10000 - 30000 psi.

[0028] In addition, the present invention provides the application of the repair and soothing composition in the preparation of cosmetics. Further, the present invention provides the application of the repair and soothing composition in the preparation of cosmetics.

[0029] Furthermore, the present invention provides a cosmetic, and the cosmetic includes the repair and soothing composition.

[0030] Preferably, the cosmetic includes any one of lotion, emulsion, cream, facial mask, essence, essential oil, and spray.

[0031] In a specific embodiment, the present invention provides a serum, which comprises the following components in mass percentages: 1-5% of the repair and soothing composition, 0.1-1% of a thickener, 0.5-5% of a humectant, 0.01-0.3% of a pH regulator, 0.5-3% of a preservative, and the balance being deionized water.

[0032] Specifically, the thickener includes at least one of polyacrylate crosspolymer-6, carbomer, carrageenan, gellan gum, xanthan gum, microcrystalline cellulose, cellulose gum, ethyl cellulose, tara gum, guar gum, acryloyldimethyltaurate / VP copolymer, and acrylate copolymer. The humectant includes at least one of glycerin, D-panthenol, vitamin B5, 1,3-butanediol, 1,2-hexanediol, 1,3-propanediol, sodium hyaluronate, tremella polysaccharide, trehalose, betaine, allantoin, and low molecular weight sodium hyaluronate. The pH regulator includes at least one of arginine, disodium ethylenediaminetetraacetate, tromethamine, and EDTA disodium. The preservative includes at least one of p-hydroxyacetophenone and polyhydric alcohol.

[0033] Compared with the prior art, the beneficial effects of the present invention are as follows: The repair and soothing composition provided by the present invention achieves rapid repair and long-term stability through a multi-dimensional synergistic mechanism, which is specifically reflected as follows: The rapid repair is reflected in that the composition for post-medical aesthetic repair and soothing provided by the present invention can relieve discomfort such as postoperative pain, burning sensation, and redness and swelling. The long-term stability is reflected in that the composition for post-medical aesthetic repair and soothing provided by the present invention can reconstruct the physical barrier and the microecological barrier, and maintain the skin homeostasis for a long time, covering the entire cycle of postoperative repair. Description of the Drawings

[0034] Figure 1 Shows the skin improvement conditions 30 minutes and 28 days after subjects used the serums prepared in Application Example 1 and Comparative Application Example 4 after photorejuvenation. Specific Embodiments

[0035] To better illustrate the purpose, technical solutions, and advantages of the present invention, the present invention will be further described below in conjunction with the drawings and specific embodiments. The purpose is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present invention. The experimental reagents and instruments designed in the embodiments and comparative examples of the present invention are all common ordinary reagents and instruments, unless otherwise specified, and can be obtained from commercial channels. In the embodiments and comparative examples, the experimental methods used are all conventional methods unless otherwise specified; and without special instructions, the raw materials used in parallel experiments are from the same batch of raw materials.

[0036] Now, the raw materials of the present invention will be further described, but are not limited to the following raw materials:

[0037] Citrus fruit extract: Purchased from Shanghai Jiaye Biotechnology Co., Ltd., with the trade name Yuefuning;

[0038] Date fruit extract: Purchased from Lucas Meyer Company, with the trade name

[0039] Gentiana root extract: Purchased from Ganeng Biotechnology (Shanghai) Co., Ltd.;

[0040] Dipotassium glycyrrhizinate: Purchased from Gansu Fanzhi Pharmaceutical Co., Ltd.;

[0041] Phytol: Purchased from BASF;

[0042] Piper methysticum root extract: Purchased from Ganeng Biotechnology (Shanghai) Co., Ltd., with the trade name Kava Calm;

[0043] Schisandra chinensis hydrolyzate: Purchased from Expanscience Laboratoires, France, with the trade name

[0044] Centella asiatica extract: Purchased from Shaanxi Xintianyu Biotechnology Co., Ltd.

[0045] Liposome-1: Self-prepared, and the preparation method is as follows:

[0046] (1) Mix soy lecithin, cholesterol, phytol, and 1,2-dioleoyl-3-trimethylammonium chloride alanine and dissolve them in solution A. After rotary evaporation at 50 °C for 30 min, a lipid film is obtained, and the rotation speed of the rotary evaporation is 150 rpm;

[0047] The ratio of soy lecithin:cholesterol:phytol:1,2-dioleoyl-3-trimethylammonium chloride alanine is 60:30:5:5; solution A is a mixed solution of chloroform and ethanol, and the weight ratio of chloroform to ethanol is 2:1; the mass-volume ratio of the mixture of soy lecithin, cholesterol, phytol, and 1,2-dioleoyl-3-trimethylammonium chloride alanine to solution A is 100 mg:4 mL;

[0048] (2) Add solution B to the lipid film in step (1), and perform ultrasonic hydration at a power of 150 w and 60 °C for 30 min and homogenize at a pressure of 20000 psi to obtain the liposome; solution B is a phosphate buffer solution containing dipotassium glycyrrhizinate, and the mass concentration of solution B is 2 mg / mL.

[0049] Among them, the weight ratio of phytol to dipotassium glycyrrhizinate in the raw materials is 5:8.

[0050] Liposome - 2: Self - prepared. Compared with the preparation method of Liposome - 1, only the mass concentration and addition amount of Solution B in step (2) are different. The mass concentration of Solution B is 2 mg / mL. Among them, the weight ratio of phytol to dipotassium glycyrrhizinate in the raw materials is 5:12.

[0051] Liposome - 3: Self - prepared. Compared with the preparation method of Liposome - 1, only the mass concentration and addition amount of Solution B in step (2) are different. The mass concentration of Solution B is 2 mg / mL. Among them, the weight ratio of phytol to dipotassium glycyrrhizinate in the raw materials is 5:20.

[0052] Liposome - 4: Self - prepared. Compared with the preparation method of Liposome - 1, only the raw material dosage in step (1) is different. The ratio of soybean lecithin: cholesterol: phytol: 1,2 - dioleoyl - 3 - trimethylammonium chloride alanine is 50:30:10:10. Among them, the weight ratio of phytol to dipotassium glycyrrhizinate in the raw materials is 5:4.

[0053] Liposome - 5: Self - prepared. Compared with the preparation method of Liposome - 1, only the mass concentration and addition amount of Solution B in step (2) are different. The mass concentration of Solution B is 5 mg / mL. Among them, the weight ratio of phytol to dipotassium glycyrrhizinate in the raw materials is 5:25.

[0054] Examples and Comparative Examples

[0055] Examples and comparative examples of the present invention provide a repairing and soothing composition. The components and weight parts of the repairing and soothing composition are selected as shown in Table 1 - 2. The preparation method of the repairing and soothing composition is as follows: Mix each component evenly to obtain the repairing and soothing composition.

[0056] Among them, in Comparative Example 6, phytol and dipotassium glycyrrhizinate in the raw materials are not prepared into liposomes, and each liposome component is added separately and directly mixed evenly with the remaining raw materials to obtain the repairing and soothing composition.

[0057] Table 1

[0058]

[0059]

[0060] Table 2

[0061]

[0062]

[0063] Application Examples

[0064] The compositions for post - aesthetic - procedure repair and soothing prepared in the examples and comparative examples were formulated into essence liquids as shown in Table 3. Among them, the compositions in Application Examples 1 - 9 used the compositions prepared in Examples 1 - 9 respectively; the compositions in Comparative Application Examples 1 - 9 used the compositions prepared in Comparative Examples 1 - 9 respectively. For example, the composition used in Application Example 1 was the composition in Example 1, the composition used in Application Example 8 was the composition in Example 8, the composition used in Application Example 10 was the composition in Example 1, and so on; the composition used in Comparative Application Example 1 was the composition prepared in Comparative Example 1.

[0065] Table 3

[0066]

[0067]

[0068] This application provides a method for preparing an essence liquid, and the preparation method includes the following steps:

[0069] (1) Mix a humectant and a thickener with water and stir. After heating to 85 °C, homogenize at a rotation speed of 1300 rpm for 4 min to obtain a pre - prepared Component A;

[0070] (2) Mix preservatives and heat to 60 °C to melt to obtain a pre - prepared Component B;

[0071] (3) After cooling the pre - prepared Component A to 60 °C, add the pre - prepared Component B and stir - mix at a rotation speed of 300 rpm. Subsequently, cool to below 45 °C, add the composition and continue stirring for 8 min. Finally, add a pH regulator to adjust the pH to 5.5 - 6.0, then stop stirring, discharge the material, and obtain the essence liquid.

[0072] Performance Test - 1: In vitro experiments verified that the composition can improve postoperative burning and itching.

[0073] Aesthetic medical operations (such as laser, microneedle, etc.) will damage the skin barrier and trigger an acute inflammatory response, resulting in redness, swelling, pain, and excessive release of pro - inflammatory factors (such as TNF - α, IL - 6, etc.). The persistence of inflammation will exacerbate oxidative stress and cell damage, prolong the recovery period, and even cause post - inflammatory hyperpigmentation (PIH) or scar hyperplasia. By inhibiting the synthesis and release of inflammatory mediators, the inflammatory cascade reaction can be blocked, reducing the oxidative damage of free radicals to new tissues, and at the same time alleviating the irritation of nerve endings, thereby rapidly improving discomforts such as postoperative burning and itching. The better the effect of the composition in inhibiting inflammation, the better the corresponding soothing and repairing effect.

[0074] The specific method is as follows:

[0075] (1) Sample preparation: Weigh 1.00 g of the repair and soothing compositions prepared in Examples 1-9 and Comparative Examples 1-9 respectively, dissolve them in PBS buffer (pH 6.8), and make up the volume to 100 mL to obtain a sample working solution with a concentration of 1.0%.

[0076] (2) Cell treatment: Seed RAW264.7 cells in the logarithmic growth phase into a 24-well plate at a cell density of 2×10 5 cells / mL, 0.5 mL per well. After plating, mark the upper cover of the cell plate and incubate it in a carbon dioxide incubator for 24 h.

[0077] (3) Sample treatment: After 24 h of cell incubation, aspirate the supernatant. The model group is LPS (1 μg / mL) + culture medium, a total of 200 μL; the sample group is cell culture medium containing 1.0% of the composition of the example or comparative example + LPS (1 μg / mL), a total of 200 μL.

[0078] (4) Detection method: After treating the cells with the sample for 24 h, use a pipette to aspirate the supernatant into a centrifuge tube, centrifuge at 1000 r / min for 5 min, discard the precipitate, aspirate the supernatant, and use a mouse tumor necrosis factor-α ELISA detection kit (Xinbosheng, EMC102a.96) to detect the change in the inflammatory factor in the supernatant.

[0079] The calculation method is as follows:

[0080] TNF-α inhibition rate % = |(TNF-α amount 模型组 - TNF-α amount 实验组 )| / TNF-α amount 模型组 × 100%; among them, the higher the TNF-α inhibition rate %, the stronger the anti-inflammatory ability.

[0081] The test results are shown in Table 4.

[0082] Table 4

[0083]

[0084]

[0085] As can be seen from the above table, the repair and soothing compositions prepared in the examples of the present invention can well inhibit the synthesis and release of inflammatory mediators, and can well improve postoperative burning and itching.

[0086] Comparing Examples 1 - 5, it can be seen that when the repair and soothing composition of the present invention is within a specific weight range, such as "0.3 - 0.8 parts of citrus fruit extract, 0.2 - 0.3 parts of date fruit extract, 0.1 - 0.3 parts of gentian root extract, 0.3 - 0.5 parts of liposome", it can better inhibit the synthesis and release of inflammatory mediators and significantly improve postoperative burning and itching.

[0087] Among them, comparing Example 1 and Example 3, it can be seen that the specific weight ratios of the citrus fruit extract, date fruit extract, and gentian root extract do not show obvious differences in the cell experiment. Comparing Examples 1 - 3, it can be seen that when the weight percentage of the liposome is 30 - 40%, it can better inhibit the synthesis and release of inflammatory mediators and significantly improve postoperative burning and itching.

[0088] Comparing Examples 1, 6 - 9, it can be seen that the weight ratio of phytol and dipotassium glycyrrhizinate, the raw materials for preparing the liposome, has a certain impact on the synthesis and release of inflammatory mediators. When the weight ratio of phytol and dipotassium glycyrrhizinate is 5:(8 - 12), the inhibitory effect on the synthesis and release of inflammatory mediators is the best.

[0089] Comparing Examples 1, Comparative Examples 1 - 9, it can be seen that when one or two components are not added, or when the other components are used to make up the amount while not adding them, the effects of the present invention cannot be achieved; when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained. Among them, comparing Example 1 and Comparative Example 6, it can be seen that in Comparative Example 6 compared with Example 1, when not prepared into liposomes, it is found in the actual experiment that when phytol and dipotassium glycyrrhizinate are not prepared into liposomes, dipotassium glycyrrhizinate and phytol are partially degraded and ineffective, and long - term stability cannot be achieved. Among them, comparing Example 1 and Comparative Example 7, in Comparative Example 7, the kava kava root extract is used to replace the citrus fruit, and it can also strongly inhibit TNF - α, but according to the subsequent data of the skin cold - sensation channel, the kava kava root extract cannot activate the TRPM8 skin cold - sensation channel. In Comparative Examples 8 - 9, the schisandra chinensis hydrolyzate and asiatic pennywort extract are used to replace some components of the present application respectively, and the synergy with the rest cannot be achieved, and the effect of inhibiting the synthesis and release of inflammatory mediators is poor.

[0090] Performance Test - 2 In vitro experiments verify that the composition can activate the TRPM8 skin cold - sensation channel.

[0091] As a temperature-sensitive ion channel, after being activated, TRPM8 promotes the influx of calcium ions and triggers the transmission of the cooling signal to the central nervous system, directly inhibiting the thermal pain conduction mediated by TRPV1 and forming an antagonistic effect between cold and heat signals. This activation can not only reduce inflammatory reactions such as postoperative erythema and edema. At the same time, the continuous activation of TRPM8 can maintain a low-temperature state in the epidermal microenvironment, reduce oxidative stress damage and promote the migration of keratinocytes, accelerating the barrier repair process. Therefore, the better the effect of the composition in activating the expression of the TRPM8 gene, the more it can soothe the skin after medical aesthetics and play a cold compress analgesic effect.

[0092] The specific method is as follows:

[0093] (1) Sample preparation: Weigh 1.00 g of the repair and soothing compositions prepared in Examples 1-9 and Comparative Examples 1-9 respectively, dissolve them in PBS buffer (pH 6.8), and make up the volume to 100 mL to obtain a sample working solution with a concentration of 1.0%.

[0094] (2) Cell culture: Inoculate the HaCaT cell suspension (purchased from Guangzhou Customs and cryopreserved by the laboratory) into a 96-well cell culture plate at a density of 2000 cells / well, add 100 μL of DMEM high-glucose culture medium (Gibco) containing 10% fetal bovine serum to each well, and culture for 24 h.

[0095] (3) Sample treatment: After 6 h of cell incubation, aspirate the supernatant. The blank group is 200 μL of DMEM high-glucose culture medium; the sample group is a total of 200 μL of cell culture medium containing 1.0% of the composition of the example or comparative example.

[0096] (4) Collect the cells, extract the total RNA of the cells with TRIzol reagent, and then synthesize cRNA with the reverse transcription kit PrimeScript RT; then use the CFX96 Touch Real-Time PCR detection system (Bio-Rad, Hercules, CA, USA) for relative quantitative analysis of the mRNA expression level. The experimental conditions are: initial denaturation at 95 °C for 5 min, and then 40 cycles of 95 °C / 15 s, 60 °C / 30 s, and 72 °C / 30 s; after normalizing to the GAPDH transcript level, use the 2-ΔΔCt method to calculate the relative cDNA level.

[0097] The calculation method is as follows:

[0098] The change rate of the TRPM8 gene % = |(the relative cDNA level of each gene in the blank group - the relative cDNA level of each gene in the sample group)| / the relative cDNA level of each gene in the blank group × 100%

[0099] The results obtained are shown in Table 5.

[0100] Table 5

[0101]

[0102]

[0103] As can be seen from the above table, the repair and soothing composition prepared in the embodiments of the present invention can well activate the TRPM8 skin cold-sensing channel.

[0104] From the comparison of Examples 1-5, it can be seen that the repair and soothing composition of the present invention can better activate the TRPM8 skin cold-sensing channel when in a specific weight part range, such as "0.3-0.8 parts of citrus fruit extract, 0.2-0.3 parts of date fruit extract, 0.1-0.3 parts of gentian root extract, 0.3-0.5 parts of liposome". Among them, from the comparison of Example 1 and Example 3, it can be seen that when the specific weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract is in a specific range, the TRPM8 skin cold-sensing channel can be better activated. When the weight percentage content of the liposome is 30-40%, the TRPM8 skin cold-sensing channel can be better activated. In cell experiments, from the comparison of Examples 1, 6-9, it can be seen that the weight ratio of phytol and dipotassium glycyrrhizinate, the raw materials for preparing the liposome, has almost no effect on the activation of the TRPM8 skin cold-sensing channel. The reason may be that in this cell experiment process, the activation of the TRPM8 skin cold-sensing channel is a short-term process, and as long as it is prepared into a liposome, there is a good effect of activating the TRPM8 skin cold-sensing channel.

[0105] From the comparison of Example 1 and Comparative Examples 1-9, it can be seen that when one or two components are not added, or when other component parts are supplemented while not adding, the effects of the present invention cannot be achieved; when replaced with other similar components, the corresponding effects of the present invention cannot be obtained. Among them, from the comparison of Example 1 and Comparative Example 6, it can be seen that compared with Example 1, in Comparative Example 6, when not prepared into a liposome, it is found in actual experiments that phytol and dipotassium glycyrrhizinate cannot synergistically activate the TRPM8 skin cold-sensing channel with other components when not prepared into a liposome. Among them, from the comparison of Example 1 and Comparative Example 7, in Comparative Example 7, the kava root extract replaces the citrus fruit, and the kava root extract cannot activate the TRPM8 skin cold-sensing channel. In Comparative Examples 8-9, the schisandra chinensis hydrolyzate and centella asiatica extract are used to replace some components of the present application respectively, and the synergy with the rest cannot be achieved, and the effect of activating the TRPM8 skin cold-sensing channel is poor.

[0106] Performance test - 3 Human skin patch test.

[0107] Because the skin after medical aesthetics is very sensitive and has higher requirements for the safety and mildness of repair and soothing products, we have tested the safety of skin care products. 32 volunteers aged 20 - 50 were recruited and the closed patch test method was used. Since the skin on the arm is thicker and tougher than that on the face, in order to simulate the damaged skin state of the subjects after medical aesthetics, we pre - used medical 3M tape to perform 5 tearing operations on the arms of the subjects, causing a certain degree of damage to the arm barrier of the subjects. An equal amount (0.020 mL - 0.025 mL) of the test samples (serums prepared from the application examples, comparative application examples, and blank application examples) was placed in a specific 10 - hole patch tester. The test samples could exactly use two 10 - hole patch testers. The 32 volunteers were divided into two groups of 16 each, and one of the patch testers was randomly tested on the left arm of the volunteers. The specific operation was as follows: it was applied to the arm of the volunteer with a low - allergenic tape and gently pressed to make it evenly adhere to the skin and left for 24 h; the blank control group was distilled water, and the blank application example was the serum without the composition. After 24 h, the patch tester was removed, and after another 24 h, the skin reaction was observed and the results were recorded. The adverse reaction grades of the skin are shown in Table 6 below, and the test results are shown in Table 7 below.

[0108] Table 6

[0109]

[0110] Table 7

[0111]

[0112]

[0113] As can be seen from the above table, the serums prepared from the application examples, comparative application examples, and blank application examples of the present invention all showed negative reactions.

[0114] Performance Test - 4 Repair effect of the composition on people after medical aesthetics.

[0115] Experimental method: According to the "Technical Specifications for Safety of Cosmetics" (2015), subjects aged 20-50 were selected to conduct the post-operative repair and soothing experiment of photorejuvenation. 12 groups were randomly divided into 3 groups, each with 3 people. All subjects clarified the experimental requirements and agreed to undergo photorejuvenation on the face and complete the test. The photorejuvenation M22 device was selected, and the hospital operating technicians would adjust the intensity of use according to the specific facial condition and different areas of the subjects. After the subjects measured the basic value after surgery, they used the sample and measured the data of using the sample for 30 minutes. After that, the volunteers used the sample once in the morning and evening every day, and the whole face (the samples were the essence prepared by application examples 1-5, 7-9, and the comparison application examples 4, 6, 9 and the blank application example), and visited the 14th and 28th days to collect data. After the visit, the volunteers sat in an air-conditioned room with a temperature of 21±1℃ and a humidity of 50±10% for 30 minutes. The researchers used skin water loss to test the transepidermal water loss value TEWL of the zygomatic bone. Photorejuvenation is a non-invasive medical aesthetic project, so redness cannot be observed with the naked eye after the operation. Therefore, the VISA-CR instrument is used to photograph the red area of the subject's face and quantify the degree and area of redness. Specifically, VISIA-CR can filter out the light reflected from the skin surface in the cross-polarized light mode, capture the distribution of microvessels and the inflammatory state within 2 mm under the skin, and visualize the deep redness. Use Image-Pro Plus software to analyze the red area image and extract the a* value (red-green axis chromaticity coordinates). The higher the a* value, the more severe the skin redness, and the red pixel area divided by the total analyzed area gives the red area.

[0116] The barrier repair ability of the composition after medical beauty treatment is represented by the improvement of TEWL value, and the soothing ability is represented by the degree of redness and the area of redness. The formula is as follows:

[0117] Improvement rate % = |X 使用前均值 -X 使用后均值 | / X 使用前均值 ×100%;

[0118] The results are shown in Table 8.

[0119] Table 8

[0120]

[0121]

[0122] It can be seen from the above table that after the repairing and soothing composition prepared in the embodiment of the present invention is prepared into an essence, it can well improve the barrier repair and soothing effects after photorejuvenation. Figure 1For the skin improvement of the subjects after photon rejuvenation, 30 minutes and 28 days after using the essence prepared in Application Example 1 and Comparative Application Example 4, it can be seen that after the repair and soothing composition prepared in the embodiment of the present invention is prepared into an essence, it can well improve the barrier repair and soothing effects after photon rejuvenation.

[0123] For the barrier repair and soothing effects 30 minutes later, it can be seen from the comparison of Application Examples 1-5 that the repair and soothing composition of the present invention can better improve the barrier repair ability and soothing ability when in a specific weight part range such as "0.3-0.8 parts of citrus fruit extract, 0.2-0.3 parts of date fruit extract, 0.1-0.3 parts of gentian root extract, 0.3-0.5 parts of liposome". Among them, from the comparison of Application Example 1 and Application Example 3, it can be seen that when the specific weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract is in a specific range, and the weight percentage content of the liposome is 30-40%, the barrier repair ability and soothing can be better improved. From the comparison of Application Example 1 and Application Examples 7-9, it can be seen that the weight ratio of phytol and dipotassium glycyrrhizinate, the raw materials for preparing the liposome, has little effect on the barrier repair ability and soothing ability. The reason may be that the activation of the TRPM8 skin cold-sensing channel is a short-term process. As long as it is prepared into a liposome, it can well improve the barrier repair and soothing, and the influence of the raw materials of the liposome on the barrier repair and soothing at 30 minutes is not obvious.

[0124] From the comparison of Application Example 1, Comparative Application Examples 4, 6, and 9, it can be seen that when one or two components are not added and the number of other components is supplemented, and when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained. Among them, from the comparison of Application Example 1 and Comparative Application Example 6, it can be seen that when not prepared into a liposome, in actual experiments, it is found that when dipotassium glycyrrhizinate and phytol are not prepared into a liposome, dipotassium glycyrrhizinate and phytol degrade and fail, and cannot cooperate well with other components. Among them, in Comparative Application Example 9, the centella asiatica extract is used to replace the gentian root extract of the present application, and the cooperation with components such as citrus fruit extract and date fruit extract cannot be achieved.

[0125] For the barrier repair and soothing effects at 14 days and 28 days, it can be seen from the comparison of Application Examples 1-5 that the repair and soothing composition of the present invention can better provide the barrier repair and soothing effects when in a specific weight part range such as "0.3-0.8 parts of citrus fruit extract, 0.2-0.3 parts of date fruit extract, 0.1-0.3 parts of gentian root extract, 0.3-0.5 parts of liposome". Among them, from the comparison of Application Example 1 and Application Example 3, it can be seen that the specific weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract has no obvious difference in the effects at 14 days and 28 days. From the comparison of Examples 1-3, it can be seen that when the weight percentage content of the liposome is 30-40%, the barrier repair and soothing effects are better.

[0126] It can be seen from the comparison between Application Example 1 and Application Examples 6-9 that the weight ratio of phytol and dipotassium glycyrrhizinate, which are the raw materials for liposome preparation, has a certain impact on barrier repair and soothing. When the weight ratio of phytol to dipotassium glycyrrhizinate is 5:(8-12), the barrier repair and soothing effects are the best. It can be seen from the comparison between Application Example 1 and Comparative Application Examples 4, 6, and 9 that when one or two of the components are not added and the number of other components is used to make up, or when other similar components are used for substitution, the corresponding effects of the present invention cannot be achieved. Among them, it can be seen from the comparison between Application Example 1 and Comparative Application Example 6 that without preparing into liposomes, dipotassium glycyrrhizinate and phytol will degrade and become ineffective, and long-term stability cannot be achieved.

[0127] 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. A repair and soothing composition, characterized in that, It comprises the following components in parts by weight: 0.1 - 1 part of citrus fruit extract, 0.1 - 0.5 part of date fruit extract, 0.05 - 0.5 part of gentian root extract, and 0.1 - 1 part of liposome; the liposome comprises dipotassium glycyrrhizinate and phytol.

2. The repair and soothing composition according to claim 1, characterized in that, It comprises the following components in parts by weight: 0.3 - 0.8 part of citrus fruit extract, 0.2 - 0.3 part of date fruit extract, 0.1 - 0.3 part of gentian root extract, and 0.3 - 0.5 part of liposome.

3. The repair and soothing composition according to claim 1, wherein Based on the total weight of the composition for repair and soothing, the weight percentage of the liposome is 30 - 40%.

4. The repair and soothing composition according to claim 1, wherein The weight ratio of the citrus fruit extract, date fruit extract, and gentian root extract is citrus fruit extract: date fruit extract: gentian root extract = (2 - 3):1:

1.

5. The repair and soothing composition according to claim 1, wherein The preparation method of the liposome comprises the following steps: (1) Mix soy lecithin, cholesterol, phytol, and 1,2 - dioleoyl - 3 - trimethylammonium chloride alanine and dissolve them in solution A, and obtain a lipid film after rotary evaporation; solution A is a mixed solution of chloroform and ethanol. (2) Add solution B to the lipid film in step (1), and obtain the liposome through ultrasonic hydration and homogenization; solution B is a phosphate buffer solution containing dipotassium glycyrrhizinate.

6. The repair and soothing composition according to claim 1, wherein The weight ratio of soy lecithin, cholesterol, phytol, and 1,2 - dioleoyl - 3 - trimethylammonium chloride alanine is soy lecithin: cholesterol: phytol: 1,2 - dioleoyl - 3 - trimethylammonium chloride alanine = (50 - 70):(20 - 40):(5 - 10):(5 - 10); and / or, the weight ratio of chloroform and ethanol is (2 - 3):1; and / or, the mass - volume ratio of the mixture of soy lecithin, cholesterol, phytol, and 1,2 - dioleoyl - 3 - trimethylammonium chloride alanine and solution A is (80 - 120)mg:4mL; and / or, the mass concentration of solution B is 1 - 5mg / mL; and / or, the weight ratio of phytol and dipotassium glycyrrhizinate is 5:(4 - 25).

7. The repair and soothing composition according to claim 6, wherein The temperature of the rotary evaporation is 45 - 60°C, the rotation speed of the rotary evaporation is 100 - 200rpm, and the time of the rotary evaporation is 30 - 50min; and / or, the temperature of the ultrasonic hydration is 50 - 60°C, the time of the ultrasonic hydration is 10 - 30min, and the power of the ultrasonic hydration is 100 - 200w; and / or, the pressure of the homogenization is 10000 - 30000psi.

8. Use of a composition for repair and soothing according to any one of claims 1 - 7 in the preparation of cosmetics.

9. A cosmetic, characterized in that, The cosmetics comprise a composition for repair and soothing according to any one of claims 1 - 7.

10. The cosmetic according to claim 9, characterized in that, The cosmetics comprise any one of lotion, emulsion, cream, facial mask, essence, essential oil, spray.

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