Soothing composition for people with sensitive skin as well as preparation method and application of soothing composition

Abstract

The invention relates to the technical field of relieving and repairing, in particular to a relieving composition for people with sensitive skin as well as a preparation method and application of the relieving composition. The soothing composition comprises a radix gentianae extract, hydrolyzed ginsenoside and a pine pollen extract. Full-link control is realized based on mechanism barrier-nerve blood vessel-immune inflammation formed on the back of sensitive skin, specific components are screened out for compounding, the synergistic effect of all the components is improved, the composition has excellent capabilities of improving the barrier, resisting inflammation and reducing neural immune response, the relieving effect is remarkable, and the composition has a good application prospect. And a relatively strong effect can be achieved by a relatively low addition amount.

Description

Technical Field

[0001] This invention relates to the field of soothing and repair technology, and in particular to a soothing composition for people with sensitive skin, its preparation method and application. Background Technology

[0002] Sensitive skin (SS) is a clinical manifestation of hyperresponsiveness of the skin under physiological or pathological conditions, primarily affecting the face. Its clinical manifestations include subjective symptoms such as burning, stinging, itching, and tightness when stimulated by physical, chemical, or psychological factors, with or without objective signs such as erythema, scaling, and telangiectasia. The International Forum on the Study of Itch (IFSI) defines SS as: unpleasant sensations (such as burning, pain, itching, stinging, etc.) produced by the skin upon exposure to external stimuli; the affected area may be normal or red.

[0003] The etiology and pathogenesis of sensitive skin are complex and diverse, mainly involving impaired skin barrier function, dysfunction of skin sensory nerves, skin microbiome dysbiosis, increased vascular reactivity, transcriptomics, and other aspects, often composed of multiple mechanisms working together. Furthermore, sensitive skin may be a skin condition or a manifestation of skin and extra-skin diseases.

[0004] In recent years, more and more people have been troubled by sensitive skin. Although there are many soothing and repairing products available, these products have relatively simple functions and are prone to relapse after discontinuation. The main reason is that the underlying causes of sensitive skin are complex. Previous studies have focused on one or two aspects of soothing effects, such as inhibiting the expression of inflammatory factors or reducing the neuroimmune response.

[0005] Chinese Patent CN117503664A discloses a soothing and repairing composition, which is a nano-lipid encapsulation made of phase A and phase B. Phase A is the soothing and repairing component, which includes Bletilla striata root extract, ginseng root extract, ceramide NP, gentian root extract, Centella asiatica leaf extract, and tetrahydromethylpyrimidine carboxylic acid, and specifies the weight percentage content of each raw material component. This invention's soothing and repairing composition uses ceramide NP and tetrahydromethylpyrimidine carboxylic acid, combined with a scientifically formulated blend of multiple plant extracts. The product is mild and safe, and will not cause allergies or irritation to users, making it safe to use. This product has a significant soothing and repairing effect, providing excellent relief and repair during daily use or when allergies, redness, itching, etc., and can effectively repair the skin's stratum corneum barrier and improve skin tolerance.

[0006] Chinese Patent CN118267416A discloses a composition with anti-inflammatory, redness-relieving, and soothing effects, wherein the composition contains component a and component b, wherein component a is a compound gentian extract comprising extracts of gentian, fangchi, and centella asiatica, and component b comprises hydrolyzed ginsenosides. The invention also relates to the use of the composition in the preparation of pharmaceuticals or cosmetics, and further relates to pharmaceuticals or cosmetics containing the composition.

[0007] As research progresses, it is generally accepted that the cause of sensitive skin is a complex process involving the skin barrier, neurovascular pathways, and immune inflammation. However, there is currently very little research on simultaneously improving these three pathways. Therefore, there is an urgent need to develop a soothing composition that controls the entire process to meet the soothing needs of people with sensitive skin. Summary of the Invention

[0008] In view of this, the purpose of this invention is to provide a soothing composition for people with sensitive skin, its preparation method and application. Based on the underlying mechanism of sensitive skin formation, it achieves full-link control of the barrier-neurovascular-immune inflammation, selects specific ingredients for compounding, improves the synergistic effect between the components, and has excellent ability to improve the barrier, anti-inflammation and reduce neuroimmune response. It has significant soothing effect and can exert strong efficacy with a low addition amount.

[0009] To achieve the above-mentioned objectives, the technical solution of this invention is as follows: On one hand, the present invention provides a soothing composition for people with sensitive skin, comprising gentian root extract, hydrolyzed ginsenosides and pine pollen extract.

[0010] Gentian root extract, hydrolyzed ginsenosides, and pine pollen extract can be purchased directly from commercially available products, or extracted using conventional extraction methods in the field (such as water extraction, alcohol extraction, other conventional solvent extraction, reflux extraction, ultrasonic extraction, etc., one or a combination thereof). They can also be further purified by combining conventional purification methods in the field (such as filtration, macroporous resin adsorption, alcohol precipitation, extraction, concentration, drying, etc., one or a combination thereof) to increase the content of active ingredients.

[0011] Preferably, the gentian root extract contains gentiopicroside ≥1200μg / mL; the hydrolyzed ginsenosides contain total saponins ≥40000ppm and rare ginsenoside CK ≥4000ppm; and the pine pollen extract has a purity of 20%-40%.

[0012] Preferably, in the soothing composition, the mass ratio of gentian root extract, hydrolyzed ginsenosides and pine pollen extract is 0.5-1.9:0.4-1.5:0.1-1.

[0013] As an example, the mass ratio of gentian root extract, hydrolyzed ginsenosides, and pine pollen extract can be selected from 0.5-1:0.4-1:0.1-0.5, 1-1.9:1-1.5:0.5-1, 0.5-1.5:1-1.5:0.1-0.5, 0.5-1:1-1.5:0.1-0.5, 1-1.9:0.4-1:0.5-1, etc., and more specifically, it can also be selected from 0.5:1.5:0.1, 1.9:0.4:1, 0.5:0.4:1, 1:1:0.5, etc.

[0014] More preferably, the mass ratio of the gentian root extract, hydrolyzed ginsenosides, and pine pollen extract is 0.5-1.6:0.4-1.5:0.5-1. More preferably, the mass ratio of the gentian root extract, hydrolyzed ginsenosides, and pine pollen extract is 1.4-1.6:0.4-1:0.5-1.

[0015] Most preferably, the mass ratio of the gentian root extract, hydrolyzed ginsenosides, and pine pollen extract is 1.5-1.6:0.4-1:0.5-1.

[0016] As an example, the mass ratio of the gentian root extract, hydrolyzed ginsenosides, and pine pollen extract can be selected from 1.5:1:0.5, 1.6:0.4:1, etc.

[0017] In another aspect, the present invention provides a method for preparing the above-mentioned soothing composition, comprising the step of mixing gentian root extract, hydrolyzed ginsenosides, and pine pollen extract.

[0018] The mixing can be performed using conventional techniques in the art, and can be either homogeneous (uniformly mixed) or non-homogeneous. To make the mixing more uniform, at least one of the conventional methods for promoting homogeneity, such as stirring, shaking, ultrasound, heating, and vibration, can be added.

[0019] For example, vortex oscillation can be used to promote uniform mixing, such as selecting 1000-10000 r / min and vortex oscillation for 1-20 min.

[0020] In another aspect, the present invention provides the use of the above-mentioned soothing composition in the preparation of soothing products.

[0021] Preferably, the soothing product is a cosmetic or topical medicine that has the effects of improving the skin barrier, anti-inflammation, and reducing neuroimmune response.

[0022] The cosmetics mentioned include, but are not limited to, serums, moisturizers, face creams, lotions, face masks, body lotions, facial cleansers, shower gels, sunscreens, sunscreen sprays, shampoos, conditioners, hair masks, etc.

[0023] The external medicines include, but are not limited to, tinctures, creams, ointments, sprays, and plasters.

[0024] More preferably, the soothing product is a cosmetic product that has the effects of improving the skin barrier, anti-inflammation, and reducing neuroimmune response.

[0025] As a specific example of the present invention, the cosmetic is a face mask.

[0026] In another aspect, the present invention provides a soothing cosmetic comprising the above-mentioned soothing composition.

[0027] Preferably, in the soothing cosmetic, the mass content of the soothing composition is 0.1%-5%, more preferably 1%.

[0028] As an example, the mass ratio can be 0.1%-0.5%, 0.5%-1%, 0.5%-1.5%, 1%-1.5%, 0.8%-1.2%, 0.9%-1.1%, 1%-2%, 2%-3%, 3%-4%, 4%-5%, etc., and further examples include 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, etc.

[0029] Preferably, the cosmetic further includes one or more of water, moisturizer, thickener, chelating agent, preservative, and pH adjuster.

[0030] Preferably, the moisturizer may be one or more of the following: glycerin, polyethylene glycol, 1,3-butanediol, propylene glycol, jojoba wax PEG-120 ester, sodium hyaluronate, bis-PEG-18 methyl ether dimethylsilane, ceramide NP, panthenol, oat (AVENASATIVA) β-glucan, glyceryl polyether-26, or allantoin. The thickener may be one or more of the following: carbomer, xanthan gum, carrageenan, sodium alginate, sclerotinia gum, sodium carbomer, hydroxypropyl methylcellulose, hydroxyethyl cellulose, sodium polyacrylate, acrylate / C10-30 alkanol acrylate crosspolymer, sodium polyacrylate, acrylamide dimethyl taurate ammonium / VP copolymer, hydroxyethyl acrylate / acryloyl dimethyl taurate copolymer, or carrageenan.

[0031] The chelating agent may be one or more of EDTA and its salts, tetrasodium glutamate diacetate, or sodium phytate.

[0032] The preservative may be one or more of phenoxyethanol, methylparaben, propylparaben, p-hydroxyacetophenone, capryloyl hydroxamic acid, ethylhexylglycerin, 1,2-hexanediol, 1,2-pentanediol, octyl glycol, chlorphenesin, or DMDM ​​hydantoin.

[0033] The pH adjuster may be one or more of citric acid, sodium citrate, lactic acid, sodium lactate, sodium hydroxide, potassium hydroxide, triethanolamine, arginine, or tromethamine.

[0034] More preferably, the cosmetic is a gel mask.

[0035] Finally, the present invention provides a method for preparing the above-mentioned gel mask, comprising the following steps: S1: Sprinkle the thickener evenly onto the surface of the water until it is completely wetted; S2: Add the remaining raw material of phase A and heat up; S3: Cool down, add phase B raw material, and dissolve; S4: Add C-phase raw materials; S5: Add the D phase raw material, cool down, and discharge to obtain the gel mask.

[0036] Phase A includes thickeners, humectants, and chelating agents; Phase B includes pH adjusters; Phase C includes water, humectants, and preservatives; and Phase D includes the above-mentioned soothing composition.

[0037] Preferably, in step S1, the water includes, but is not limited to, deionized water, purified water, distilled water, ultrapure water, mineral water, drinking water, etc. As a specific example of the present invention, deionized water is used.

[0038] Preferably, in step S2, the temperature for heating is 70-90°C, and more preferably 80-85°C.

[0039] Preferably, in step S3, the cooling temperature is 40-50°C, and more preferably 45°C.

[0040] Preferably, in step S4, the C phase is premixed at 40-50°C, more preferably at 45°C. Preferably, in step S4, the cooling temperature is 30-40°C, and more preferably 33°C.

[0041] The beneficial effects of this invention are as follows: (1) This invention provides a soothing composition for people with sensitive skin, its preparation method and application. Based on the formation mechanism behind sensitive skin, the barrier-neurovascular-immune inflammation is controlled in the whole chain. Specific components are screened and compounded to improve the synergistic effect between the components. It has excellent barrier-improving, anti-inflammatory and neuroimmune response-reducing abilities, and has significant soothing effects. A low amount of addition can exert a strong effect.

[0042] (2) The gentian, ginseng and pine pollen of the present invention are all plant raw materials of natural origin, which have the advantages of mildness and safety.

[0043] (3) The composition of the present invention is water-soluble and can be widely used in various products, such as facial cleanser, toner, serum, lotion, cream, shampoo, and bath products. It is stable, does not easily precipitate, and can exert strong effects with a low addition amount.

[0044] (4) The composition of the present invention is designed for people with sensitive skin and is safer and gentler. Attached Figure Description

[0045] Figure 1 Comparative Example 7 shows a typical relief diagram (the dotted line represents the quantitative area). Figure 2 This is a typical diagram of the relief effect in Example 6 (the dotted line represents the quantitative area). Detailed Implementation

[0046] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0047] The abbreviations of the terms used in this invention are as follows: iNOS: Inducible nitric oxide synthase, abbreviated as NO in this invention. Excessive NO production is closely related to inflammatory responses and is an important inflammatory mediator. Under normal circumstances, endothelial-derived NO has an inhibitory effect on inflammatory responses, while under pathological conditions, the synthesis of large amounts of NO by inducible NO synthase leads to adverse cellular reactions, exacerbates inflammatory responses, and participates in the development of various diseases.

[0048] TRPV1: Capsaicin receptor; activation of the capsaicin receptor causes intracellular calcium... 2+ Increased sensitivity can lead to neurogenic inflammation of sensitive skin, resulting in symptoms such as vasodilation, edema, papules, itching, and burning.

[0049] HIS: Histamine. Mast cells are filled with large secretory granules containing heparin, histamine, eosinophil chemotactic factors, etc. When stimulated by allergens, they release large amounts of substances from the granules, such as histamine, via exocytosis, causing an allergic inflammatory response.

[0050] FLG: Filamentin, an important factor involved in the skin barrier, whose main function is to participate in the differentiation of epidermal cells and the formation of the skin barrier. ZO-1: Tight junction protein, a key core protein of tight junction structure, plays a crucial role in maintaining cell polarity and barrier function.

[0051] In the following examples, the gentian root extract, a yellow to light brown liquid with a gentiopicrin content ≥1200 μg / ml, was purchased from Shanghai Jiayu Biotechnology Co., Ltd., catalog number JY1004-2.5. The preparation process was carried out according to Example 3 in CN 107137261 B.

[0052] Hydrolyzed ginsenosides, light yellow to yellowish-brown liquid, with a total saponin content ≥40000ppm and a rare ginsenoside CK content ≥4000ppm, purchased from Sanyi Technology (Guangzhou) Co., Ltd., product number SUNIGA24E01-3.

[0053] Pine pollen extract, a light yellow to yellowish-brown slightly viscous clear liquid, was purchased from Yantai New Era Health Industry Daily Chemical Co., Ltd., product number 20250415. Preparation process: A certain amount of non-crystallized pine pollen was taken, and 30 times its weight of 30% ethanol was added. The mixture was refluxed at 85℃ for 3 hours, filtered, and concentrated by rotary evaporation to 2.5 times the feed volume. Four times the weight of 95% ethanol was added, and the mixture was allowed to precipitate overnight. After filtration, the mixture was rotary evaporated to 2.5 times the feed volume to obtain the pine pollen concentrate. Polyols and preservatives were added to prepare a pine pollen extract with a purity of 20%-40%.

[0054] Centella asiatica extract, a pale yellow or brownish-yellow homogeneous liquid, with a total glycoside content of ≥75mg / 100ml, was purchased from Shandong Mingren Furida Pharmaceutical Co., Ltd., product number 250601.

[0055] Kava pepper extract, a clear yellow to brownish-yellow liquid with capsaicin content of 600-1200 ppm, was purchased from Konami (Guangzhou) Biotechnology Co., Ltd., product number 20240910.

[0056] Examples 1-5, Comparative Examples 1-6 A soothing composition for sensitive skin is shown in Table 1. The raw materials are prepared by weight as follows: the components are mixed according to the following mass ratio, and the mixture is vortexed at 3000 r / min for 3-5 min until it is uniformly mixed to obtain the composition.

[0057] Table 1. Component list of Examples 1-5 and Comparative Examples 1-6

[0058] Note: "-" indicates no addition.

[0059] Comparative Example 7 The difference from Example 2 is that the pine pollen extract was replaced with Centella asiatica extract, otherwise the same.

[0060] Example 6, Comparative Example 8 A soothing face mask for sensitive skin, prepared as follows: S1: Sprinkle raw material No. 2 evenly onto the surface of deionized water until it is completely wetted.

[0061] S2: Add the remaining raw materials of phase A, stir until completely dispersed, and heat to 80-85℃.

[0062] S3: Cool to 45℃, add phase B, and stir until completely dissolved.

[0063] S4: Add the C phase after it has been mixed evenly. The C phase needs to be premixed at 45°C.

[0064] S5: Add phase D, stir well, cool to 33℃, and discharge to obtain a soothing gel mask.

[0065] The raw material composition is shown in Table 2: Table 2. Soothing Facial Masks of Example 6 and Comparative Example 7

[0066] Note: "-" indicates no addition.

[0067] Effect verification 1. Cell safety concentration determination The compositions described in this invention are all compositions prepared in Examples 1-5 and Comparative Examples 1-7.

[0068] The method for determining safe cell concentrations is as follows: Log-phase cells (human immortalized keratinocytes (HaCaT), mouse mast cell tumor cells (P815), and mouse mononuclear macrophage leukemia cells (Raw264.7)) were seeded in 96-well plates and cultured for 24 h. Cells were then divided into a normal control group (cells and culture medium) and a sample group (cells, culture medium, and the composition). The old culture medium was discarded, and the cells were washed once with PBS. Culture medium was added to the control group, and the sample solution (composition + culture medium) was added to the sample group. The cells were incubated at 37°C with 5% CO2 solution. The old culture medium was discarded, and the cells were washed twice with PBS. 110 μL of culture medium containing 10% v / v CCK-8 solution was added to each well. A blank control group was also included, with only 110 μL of CCK-8 solution added to each well. After incubation at 37°C for 2 h, the absorbance (OD) at 450 nm was measured using a microplate reader, and cell viability was calculated. The highest concentration of the composition with a cell viability greater than 95% was considered the safe cell concentration for that sample.

[0069] Human immortalized keratinocytes (HaCaT) were purchased from Hunan Fenghui Biotechnology Co., Ltd.; mouse mast cell tumor cells (P815) were purchased from Beina Chuanglian Biotechnology Co., Ltd.; and mouse mononuclear macrophage leukemia cells (Raw264.7) were purchased from the National Cell Resource Bank.

[0070] Table 3. Maximum safe concentration for cells

[0071] The final concentrations of the compositions tested in Examples 1-3 below are the corresponding concentrations in Table 3.

[0072] 2. Barrier Repair Capacity Evaluation Experimental Principle: FLG is an epidermal protein derived from the hydrolysis of profigrin giant precursor protein and is an important component of the skin barrier. ZO-1, by regulating the assembly of Claudin protein, helps form a band-like sealing structure around the top of the cell, selectively controlling the passive transport of water-soluble substances, ions, and molecules in the paracellular pathway, thus constituting a second physical barrier. Using HaCaT cells, the intracellular ZO-1 and FLG levels were measured after sample addition to evaluate the sample's effect on the barrier.

[0073] Experimental Methods: HaCaT cells in logarithmic growth phase (Hunan Fenghui Biotechnology Co., Ltd.) were collected and seeded into 24-well plates and cultured for 24 h. A normal control group (culture medium) and groups with different concentrations of samples (culture medium + different concentrations of samples) were set up, with two replicates for each group. After 24 h of culture, the cell supernatant was discarded, and the cells were gently washed twice with PBS. Cell lysis buffer (Beyotime Biotechnology Research Institute, catalog number: P0013) was added, and the cells were incubated at 4℃ for lysis. After complete lysis, cells from each well were collected into 1.5 mL centrifuge tubes, centrifuged for 5 min, and the supernatant was collected. The content of relevant proteins was detected using the ZO-1 ELISA kit (Wuhan Aidikang Biotechnology Co., Ltd., catalog number AD22587) and the FLG ELISA kit (Wuhan Aidikang Biotechnology Co., Ltd., catalog number AD22658). The protein content was determined according to the manufacturer's instructions.

[0074] Increase rate % = (Content in sample group - Content in normal group) / Content in normal group × 100%.

[0075] The results are as follows: Table 4. Evaluation of Barrier Repair Capacity

[0076] Note: Statistical analysis was performed between groups. " / " indicates no effect; "*" represents a significant difference from the normal group, ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05. The higher the expression rate of FLG and ZO-1, the stronger the repair effect on epidermal barrier function.

[0077] 3. Evaluation of the soothing effect of the neural pathway-capsaicin Experimental Principle: TRPV1 (transient receptor potential vanilloid-1) is a ligand-gated non-selective cation channel found in class C sensory afferent fibers and keratinocytes. It can be activated by various factors, including chemical substances (capsaicin), noxious heat stimulation, and acidification. Once activated, TRPV1 receptors release monovalent and divalent cations (primarily Ca2+). 2+ The substance enters the cell, triggers an action potential, and transmits this information to the higher central nervous system, producing a burning sensation. Using HaCaT for ultraviolet light damage, the anti-irritant efficacy of the sample was evaluated by detecting the intracellular calcium ion fluorescence intensity after the sample was applied to the cells.

[0078] Experimental methods: HaCaT cells in logarithmic growth phase (Hunan Fenghui Biotechnology Co., Ltd.) were collected and seeded into 96-well plates at a density of 2 × 10⁴ cells per well, and cultured for 24 h. Cells were divided into a control group (C), a model group (M), and a sample group. Cells were washed once with PBS, and except for the control group, they were exposed to ultraviolet light (UVB, 14.5 mJ / cm²). 2 Irradiate for 10 min ( / min), discard PBS, add 100 μL of complete culture medium to the control group, and add Ca-containing medium to the model group. 2+ (0.2 mg / mL) culture medium, sample groups were supplemented with Ca 2+ Culture medium and samples of different concentrations were used. After culturing for 24 hours, the cells were washed once with PBS, and except for the control group, they were irradiated with UV light for 10 min, washed twice with Hank's buffer, incubated with a calcium ion probe (Fluo-3) at 37℃ for 30 min, washed three times with Hank's buffer, and incubated at 37℃ for 30 min. The fluorescence intensity of calcium ions and cell viability were detected by a microplate reader. The calcium ion influx inhibition rate was calculated. The intracellular calcium ion detection kit was purchased from Beijing Bio-Labs Technology Co., Ltd. (Catalog No.: HR0943).

[0079] Inhibition rate % = (Calcium ion fluorescence intensity of model group - Calcium ion fluorescence intensity of sample group) / Calcium ion fluorescence intensity of model group × 100%.

[0080] The results are as follows: Table 5. Evaluation of the soothing effect of capsaicin on neural pathways.

[0081] Note: Statistical analysis was performed between groups. "*" indicates a significant difference from the model group: ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05. A higher calcium ion influx inhibition rate indicates a stronger inhibitory effect on neuroimmunity.

[0082] 4. Evaluation of anti-inflammatory pathway efficacy 4.1 NO Anti-inflammatory Experiment Experimental principle: LPS induces macrophages (RAW264.7 cells) to secrete large amounts of NO. Under acidic conditions, NO undergoes a diazo reaction with diazonium salt sulfonamide to generate a diazo compound. The latter further couples with naphthylvinyldiamine. The concentration of the product generated by this reaction has a linear relationship with the NO concentration, and has a maximum absorption peak at 540 nm.

[0083] Experimental Methods: Log-phase RAW264.7 cells (purchased from the National Experimental Cell Resource Sharing Platform) were seeded into 24-well plates and cultured for 24 h. The supernatant was carefully discarded. Wells were set up as normal control wells, LPS-induced model wells (culture medium + 1 μg / mL LPS), and sample wells containing the composition (culture medium + 1 μg / mL + composition). The corresponding solutions were added to each well, and the cells were cultured for 24 h. Following the instructions of the nitric oxide kit (purchased from Beyotime Biotechnology Co., Ltd., catalog number: S00215), 50 μL of culture supernatant was taken from each well, followed by 50 μL of reagent 1 and then 50 μL of reagent 2. OD540 was measured, and the NO concentration in each well was calculated based on the standard curve.

[0084] Formula for calculating the percentage decrease in NO: NO reduction percentage % = (NO concentration in model wells - NO concentration in sample wells) / NO concentration in model wells × 100%.

[0085] 4.2 HIS Anti-inflammatory Experiment Experimental Principle: Compound 48 / 80 trihydrochloride (C48 / 80) is a mixture of the condensation products of N-methyl-p-methoxyphenethylamine and formaldehyde. It is used as a classic mast cell activator for IgE-independent G proteins, as well as a histamine releaser and mast cell degranulation agent. C48 / 80 was used to induce degranulation in mouse mast cell tumor cells (P815 cells). The histamine (HIS) content in the cell culture supernatant was measured to evaluate the inhibitory effect of the sample on mast cell degranulation.

[0086] Experimental Methods: P815 cells in logarithmic growth phase were selected, washed three times with PBS, digested with trypsin, collected, and counted. 2 × 10⁵ cells were seeded per well in 48-well plates and cultured for 24 h. After adding the sample and culturing for 2 h, C48 / 80 (30 μg / mL) was added for stimulation for 1 h. The supernatant was collected, and the HIS content was detected according to the histamine detection kit (purchased from Shanghai Zhuocai Biotechnology Co., Ltd., catalog number: ZC-55834-J). The formula for calculating the HIS inhibition rate of the sample is as follows: Inhibition rate % = (Model group content - Sample group content) / Model group content × 100% The results are as follows: Table 6. Anti-inflammatory evaluation results

[0087] Note: Statistical analysis was performed between groups. "*" indicates a significant difference from the model group: ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05. In the cellular inflammation model, the higher the percentage reduction relative to the model group, the stronger the anti-inflammatory ability.

[0088] Conclusion: Compared with the comparative example, Examples 1-5 significantly increased the expression of FLG and ZO-1 barrier-related proteins, thereby repairing the skin barrier function. Compared with the comparative example, Examples 1-5 significantly inhibited the influx of calcium ions induced by capsaicin, thereby reducing the neuroimmune response. Compared with the comparative example, Examples 1-5 significantly inhibited the release of NO and HIS, thereby reducing inflammation. In summary, Examples 1-5 achieved synergistic effects in three aspects: barrier repair, neuroimmune response, and immune inflammation, greatly improving the soothing efficacy. Among them, Examples 2 (the mass ratio of gentian root extract, hydrolyzed ginsenosides, and pine pollen extract was 1.5:1:0.5) and Examples 3 (the mass ratio of gentian root extract, hydrolyzed ginsenosides, and pine pollen extract was 1.6:0.4:1) showed the best overall performance. Therefore, Example 2 will be applied to the development of soothing mask products to verify the soothing efficacy of the finished product.

[0089] 5. Reviews of Soothing Face Masks 5.1 Evaluation of the soothing effect of the soothing mask Experimental Principle: Sodium lauryl sulfate (SLS) may trigger a stimulating response in the organism. The stimulant enters the zebrafish, inducing an inflammatory response, causing neutrophils to initiate an immune response, migrate to the skin epidermis, and aggregate. The change in the number of skin neutrophils before and after treatment with the transgenic green fluorescent neutrophil strain zebrafish (MPX) was used to detect whether the sample had a soothing effect.

[0090] Experimental methods: Zebrafish were randomly selected into 6-well plates, 15 fish per well; a zebrafish skin inflammation model was established by water-soluble administration of SLS; samples were administered water-solublely, and a normal control group and a model control group were set up, with a volume of 3 ml per well; the samples were incubated at 28℃ in the dark for 18 h; 10 zebrafish from each experimental group were randomly selected and photographed under a fluorescence microscope, and data were analyzed and collected using advanced image processing software. The number of neutrophils (N) in the zebrafish skin was analyzed, and the soothing effect of the sample was calculated according to the formula to determine whether it has a soothing effect.

[0091] The zebrafish efficacy test results for Comparative Example 8 and Example 6 are shown below. Figure 1 , Figure 2 .

[0092] Soothing efficacy % = (Model control group N - Sample group N) / (Model control group N - Normal control group N) × 100% The results are shown in Table 7: Table 7. Evaluation Results of Soothing Mask Efficacy

[0093] The data in Table 7 show that the efficacy of Example 6 is significantly better than that of Comparative Example 8, indicating that 1% of the soothing and anti-allergy combination (Example 2) can significantly reduce the number of neutrophils in zebrafish, and the effect is significantly better than that of kava pepper extract, a soothing and anti-allergy ingredient in the industry.

[0094] 5.2 Safety Evaluation of He Yan Soothing Facial Mask Safety tests were conducted on the soothing mask of Example 6. The specific test methods are as follows: Number of participants: 30, aged 24-48, who met the selection criteria for subjects in human skin patch tests as specified in the "Cosmetic Safety Technical Specifications" (2015 edition).

[0095] Test Method: Using qualified patch testing equipment, a closed patch test method was employed. Approximately 0.020-0.025 ml of the soothing facial mask was placed in the patch testing device, and medical tape was used to apply it to the flexor side of the subject's forearm. The device was gently pressed with the palm of the hand to ensure even application to the skin. After 24 hours, the patch testing device was removed. Skin reactions were observed at 0.5, 24, and 48 hours after removal. The results were recorded according to the skin reaction grading standards in the "Cosmetic Safety Technical Specifications" (2015 edition). Table 8 shows the grading standards for adverse skin reactions.

[0096] Table 8. Grading Criteria for Adverse Skin Reactions

[0097] The results of the human skin occlusive patch test showed that 0 out of 30 people had a positive reaction. According to the "Cosmetic Safety Technical Specifications" (2015 edition), the test substance will not cause adverse skin reactions in this batch of subjects.

[0098] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A soothing composition for sensitive skin population, characterized in that, It includes gentian root extract, hydrolyzed ginsenosides, and pine pollen extract.

2. The soothing composition of claim 1, wherein, The gentian root extract contains gentiopicroside ≥1200μg / mL; the hydrolyzed ginsenosides contain total saponins ≥40000ppm and rare ginsenoside CK ≥4000ppm; the pine pollen extract has a purity of 20%-40%.

3. The soothing composition of claim 1, wherein, In the soothing composition, the mass ratio of gentian root extract, hydrolyzed ginsenosides, and pine pollen extract is 0.5-1.9:0.4-1.5:0.1-1, preferably 0.5-1.6:0.4-1.5:0.5-1, more preferably 1.4-1.6:0.4-1:0.5-1, and most preferably 1.5-1.6:0.4-1:0.5-1.

4. Process for the preparation of the soothing composition according to any one of claims 1 to 3, characterized in that, include: The step involves mixing gentian root extract, hydrolyzed ginsenosides, and pine pollen extract.

5. The use of the soothing composition according to any one of claims 1-3 in the preparation of soothing products.

6. Use according to claim 5, characterized in that, The soothing product is a cosmetic or topical medicine that has the effects of improving the skin barrier, reducing inflammation, and reducing neuroimmune response.

7. A soothing cosmetic product, characterized by, The soothing composition includes any one of claims 1-3, preferably, the mass content of the soothing composition in the soothing cosmetic is 0.1%-5%, most preferably 1%.

8. The soothing cosmetic product of claim 7, wherein, It also includes one or more of the following: water, humectant, thickener, chelating agent, preservative, and pH adjuster; Preferably, the moisturizer is selected from one or more of glycerin, polyethylene glycol, 1,3-butanediol, propylene glycol, jojoba wax PEG-120 ester, sodium hyaluronate, bis-PEG-18 methyl ether dimethylsilane, ceramide NP, panthenol, oat (AVENA SATIVA) β-glucan, glyceryl polyether-26, or allantoin. The thickener is selected from one or more of the following: carbomer, xanthan gum, carrageenan, sodium alginate, sclerotinia gum, sodium carbomer, hydroxypropyl methylcellulose, hydroxyethyl cellulose, sodium polyacrylate, acrylate / C10-30 alkanol acrylate crosspolymer, sodium polyacrylate, acrylamide dimethyl taurate ammonium / VP copolymer, hydroxyethyl acrylate / acryloyl dimethyl taurate copolymer, or carrageenan. The chelating agent is selected from one or more of EDTA and its salts, tetrasodium glutamate diacetate, or sodium phytate. The preservative is selected from one or more of phenoxyethanol, methylparaben, propylparaben, p-hydroxyacetophenone, capryloyl hydroxamic acid, ethylhexylglycerin, 1,2-hexanediol, 1,2-pentanediol, octyl glycol, chlorphenesin, or DMDM ​​hydantoin. The pH adjuster is selected from one or more of citric acid, sodium citrate, lactic acid, sodium lactate, sodium hydroxide, potassium hydroxide, triethanolamine, arginine, or tromethamine.

9. The soothing cosmetic product of claim 7, wherein, The cosmetic product is a gel mask.

10. The method of preparing a soothing cosmetic product according to claim 9, characterized in that, Including the following steps: S1: Sprinkle the thickener evenly onto the surface of the water until it is completely wetted; S2: Add the remaining raw material of phase A and heat up; S3: Cool down, add phase B raw material, and dissolve; S4: Add C-phase raw materials; S5: Add phase D raw material, cool, and discharge to obtain the gel mask. A phase includes thickening agent, humectant and chelating agent; B phase includes pH regulator; C phase includes water, humectant, preservative; D phase includes the above soothing combination, Preferably, in step S2, the temperature of the heating is 70-90℃; Preferably, in step S3, the temperature of the cooling is 40-50℃; Preferably, in step S4, the C phase is pre-mixed at 40-50℃; Preferably, in step S4, the temperature of the cooling is 30-40℃.

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