Anti-inflammatory, soothing, and repairing composition, its preparation method and application

Abstract

This application discloses an anti-inflammatory, soothing, and repairing composition, its preparation method, and its application. By weight, the anti-inflammatory, soothing, and repairing composition provided in this application comprises: 10-30 parts of *Saposhnikovia divaricata* root extract; 10-30 parts of *Butterfly Pepper* extract; and 0.01-0.5 parts of sandalwood extract. Experiments have demonstrated that the compound composition containing *Saposhnikovia divaricata* root extract, *Butterfly Pepper* leaf extract, and sandalwood extract exhibits significantly superior anti-inflammatory, soothing, and repairing efficacy compared to the combined efficacy of using these extracts alone. In the anti-inflammatory, soothing, and repairing composition provided in this application, the *Saposhnikovia divaricata* root extract, *Butterfly Pepper* leaf extract, and sandalwood extract synergistically enhance each other, achieving significant anti-inflammatory, soothing, and repairing effects even at relatively low application concentrations.

Description

Technical Field

[0001] This application belongs to the field of cosmetic technology, and in particular relates to an anti-inflammatory, soothing and repairing composition, its preparation method and application. Background Technology

[0002] In recent years, with environmental changes such as air pollution, seasonal changes, and ultraviolet radiation, coupled with increasing life and work pressures, the number of allergy sufferers has increased dramatically, and skin sensitivity has become a social problem affecting human health. According to incomplete statistics, approximately 30% of the global population has symptoms of sensitive skin, and the number of people worldwide who self-report sensitive skin is increasing, with men accounting for 38% and women for 61%.

[0003] Sensitive skin refers to a hyperreactive state of the skin under physiological or pathological conditions, primarily affecting the face. It manifests as 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 causes of sensitive skin are complex, and its formation mechanism is not yet fully understood. Current research suggests contributing factors include impaired skin barrier function, epidermal inflammation, and abnormalities in the skin's sensory nervous system. These factors often reinforce each other, leading to a vicious cycle of sensitive skin problems that makes it difficult for the skin to return to normal.

[0004] Seasonal changes / alternation (heat / cold stimulation) are often the primary trigger for sensitive skin flare-ups. There is a lack of ingredients and products on the market specifically for sensitive skin caused by seasonal changes, making the development of related ingredients and products for this condition both necessary and in demand. Summary of the Invention

[0005] The purpose of this application is to provide an anti-inflammatory, soothing, and repairing composition, its preparation method, and its application.

[0006] The first aspect of this application provides an anti-inflammatory, soothing, and repairing composition, comprising, by weight:

[0007] 10-30 parts of Saposhnikovia root extract; 10-30 parts of Acer palmatum extract; 0.01-0.5 parts of sandalwood extract.

[0008] Optionally, the anti-inflammatory, soothing, and repairing composition provided in this application comprises, by weight: 20 parts of Saposhnikovia divaricata root extract; 20 parts of Acer palmatum extract; and 0.05 parts of sandalwood extract.

[0009] The second aspect of this application provides a method for preparing the anti-inflammatory, soothing, and repairing composition of the first aspect, comprising the following steps: mixing Saposhnikovia root extract, Acer palmatum extract, Sandalwood extract with maltodextrin and water, and stirring to obtain the anti-inflammatory, soothing, and repairing composition.

[0010] Furthermore, the method for preparing the anti-inflammatory, soothing, and repairing composition provided in this application further includes the following step: spray-drying the anti-inflammatory, soothing, and repairing composition to produce a powder.

[0011] The third aspect of this application provides the use of the anti-inflammatory, soothing, and repairing composition of the first aspect in the preparation of cosmetics.

[0012] Optionally, in the above applications provided in this application, cosmetics include anti-inflammatory cosmetics, soothing cosmetics, barrier repair cosmetics, or after-sun repair cosmetics.

[0013] Optionally, in the above applications provided in this application, cosmetics include cosmetics suitable for sensitive skin.

[0014] Optionally, in the above applications provided in this application, cosmetics include at least one of the following: facial mask liquid, lotion, cream, serum, toner, essence, facial cleanser, liquid foundation, foundation cream, concealer, perfume, lotion, makeup remover, blush, eyeshadow, lipstick, and blush.

[0015] The fourth aspect of this application provides a cosmetic product comprising the anti-inflammatory, soothing, and repairing composition of the first aspect of this invention.

[0016] Optionally, the cosmetic provided in this application contains, by weight percentage: 0.1% to 1% of an anti-inflammatory, soothing, and repairing composition; the remainder being a cosmetic base.

[0017] The anti-inflammatory, soothing, and repairing composition provided in this application comprises a compound of Saposhnikovia divaricata root extract, Amaranthus urticaria leaf extract, and Sandalwood extract. The inventors of this application have discovered that the composition of Saposhnikovia divaricata root extract, Amaranthus urticaria leaf extract, and Sandalwood extract exhibits significantly superior anti-allergic, soothing, and repairing efficacy compared to the sum of the anti-allergic, soothing, and repairing efficacy of using these extracts individually. Therefore, the composition containing Saposhnikovia divaricata root extract, Amaranthus urticaria leaf extract, and Sandalwood extract has a synergistic effect, achieving remarkably significant anti-allergic, soothing, and repairing efficacy even at relatively low application concentrations.

[0018] Specifically, the composition provided in this application, comprising Saposhnikovia root extract, Amaranthus urticaria extract, and Sandalwood extract, significantly inhibits the secretion of LPS-induced inflammatory factors (NO, IL-6) and the mRNA expression of UVB-induced inflammatory factors (IL-6, IL-8), exhibiting a synergistic effect, indicating that this composition has a good inhibitory effect on inflammation caused by different factors. The compound composition of Saposhnikovia root extract, Amaranthus urticaria extract, and Sandalwood extract can inhibit the expression of the cold and heat targets TRPV1, TRPV4, and TRPA1, indicating that the anti-inflammatory, soothing, and repairing composition has a certain alleviating effect on sensitive skin caused by cold and heat. The compound composition of Saposhnikovia root extract, Amaranthus urticaria extract, and Sandalwood extract can promote the mRNA expression levels of FLG, LOR, ZO-1, and CLND3, demonstrating that this composition has a certain repairing effect on the skin barrier. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the embodiments of this application will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The figure shows the experimental results of the effect of different concentrations of Saposhnikovia root extract on cell activity in Example 1;

[0021] Figure 2 The figure shows the experimental results of the effect of different concentrations of *Begonia purpurea* leaf extract on cell activity in Example 1.

[0022] Figure 3 The figure shows the experimental results of the effect of different concentrations of sandalwood extract on cell activity in Example 1;

[0023] Figure 4 The figure shows the experimental results of the effects of Saposhnikovia root extract, Amaranthus purpurea leaf extract, Sandalwood extract, and anti-inflammatory soothing repair composition on LPS-induced NO secretion in Example 2.

[0024] Figure 5 The figure shows the experimental results of the effects of Saposhnikovia root extract, Amaranthus purpurea leaf extract, Sandalwood extract, and anti-inflammatory soothing repair composition on LPS-induced IL-6 secretion in Example 2.

[0025] Figure 6 The figure shows the experimental results of the effects of the root extract of Saposhnikovia divaricata, leaf extract of Amaranthus urinaria, sandalwood extract, and anti-inflammatory soothing and repairing composition on UVB-induced IL-6 secretion in Example 3.

[0026] Figure 7 The figure shows the experimental results of the effects of the root extract of Saposhnikovia divaricata, leaf extract of Amaranthus urinaria, sandalwood extract, and anti-inflammatory soothing and repairing composition on UVB-induced IL-8 secretion in Example 3.

[0027] Figure 8 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 4 on the expression level of TRPA1 mRNA induced by cold stimulation.

[0028] Figure 9 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 5 on the expression level of TRPV1 mRNA induced by heat stimulation.

[0029] Figure 10 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 6 on the expression level of TRPV4 mRNA induced by UVB stimulation.

[0030] Figure 11 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 7 on the expression level of FLG mRNA induced by UVB.

[0031] Figure 12 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 7 on the expression level of LOR mRNA induced by UVB.

[0032] Figure 13 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 8 on the expression level of ZO-1 mRNA induced by UVB.

[0033] Figure 14 The figure shows the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition in Example 8 on the expression level of CLND3 mRNA induced by UVB. Detailed Implementation

[0034] The features and exemplary embodiments of various aspects of this application will be described in detail below. To make the objectives, technical solutions, and advantages of this application clearer, the application will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are intended only to explain this application and not to limit it. For those skilled in the art, this application can be implemented without some of these specific details. The following description of the embodiments is merely to provide a better understanding of this application by illustrating examples.

[0035] It should be noted that, in this document, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising..." does not exclude the presence of additional identical elements in the process, method, article, or apparatus that includes said element.

[0036] It should be noted that the terms "upper", "lower", "front", "rear", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of description and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this patent.

[0037] It should also be noted that, unless otherwise explicitly specified and limited, the terms "set up," "install," "connect," and "link" should be interpreted broadly. For example, they can refer to fixed connections, detachable connections, or integral connections; they can refer to mechanical connections or electrical connections; they can refer to direct connections or indirect connections through an intermediate medium; and they can refer to the internal connection between two components. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.

[0038] Skin is easily affected by external factors such as light, heat, air pollution, climate, and bacterial infection, or by internal factors such as endocrine disorders, which can cause symptoms such as dry skin, allergies, and even aging, causing great distress to people's lives. Skin sensitivity is receiving increasing attention.

[0039] The inventors of this application have noted that TRPV1 can be activated not only by thermal stimulation above 43°C and pH changes, but also by chemical substances such as capsaicin, thereby inducing Ca2+ activation in sensory neurons. 2+Influx of toxins activates TRPV1 on sensory neurons, producing sensations of pain, burning, and itching. This triggers the release of pro-inflammatory mediators such as substance P (SP) and calcitonin gene-related peptide (CGRP), mediating neurogenic inflammation and ultimately leading to various skin diseases and pruritus. TRPV4 is activated at temperatures above 27°C, under mechanical stimulation, hypotonic conditions, and with the presence of arachidonic acid metabolites. It is associated with acute and chronic pruritus, and TRPV4 on keratinocytes also participates in the formation and repair of the skin barrier. Besides transmitting pruritus, TRPV4 can also trigger hyperalgesia, responding to acute UVB exposure. TRPA1 is associated with temperature perception. TRPA1, along with TRPM3 and TRPV1, can transmit pain caused by high temperatures. TRPA1 is also the primary pathway for cold-sensitive pain receptors, participating in inflammation and neuropathic pain associated with cold-induced abnormal pain. Oxidative free radicals generated by cold stimulation can activate TRPA1, causing hyperalgesia.

[0040] Based on this, this application targets TRPA1, TRPV1 and TRPV4, and screens and provides a naturally derived anti-inflammatory, soothing and repairing composition that has a good relieving effect on sensitive skin caused by seasonal changes / changes. It can be used as an active additive in cosmetic formulations to play a preventive and soothing role.

[0041] In a first aspect, embodiments of this application provide an anti-inflammatory, soothing, and repairing composition, comprising, by weight: 10-30 parts of Saposhnikovia divaricata root extract; 10-30 parts of Amaranthus urinaria extract; and 0.01-0.5 parts of sandalwood extract.

[0042] In some embodiments of this application, the anti-inflammatory, soothing, and repairing composition comprises, by weight: 20 parts of Saposhnikovia root extract; 20 parts of Amaranthus urinaria extract; and 0.05 parts of sandalwood extract.

[0043] Saposhnikovia root contains many coumarin and chromogen compounds. In in vitro fat cell hydrolysis experiments, Saposhnikovia root extract promoted fat hydrolysis and can be used in weight loss products; it has an scavenging effect on various free radicals and can be used as an antioxidant; it has a certain inhibitory effect on melanin and can be used in whitening products; it can shrink pores and can be used as an astringent; it also has antibacterial and anti-inflammatory effects.

[0044] *Petasites hybridus*, also known as purple petastrea, is a plant used medicinally in Europe for hundreds of years. Sesquiterpenes are considered its main active ingredients. Modern pharmacological studies have shown that sesquiterpenes such as petasin and S-petasin have antihistamine effects, inhibit leukotriene synthesis, and reduce intracellular calcium ion concentration, thereby achieving anti-inflammatory and antispasmodic effects. Preliminary in vitro pharmacological experiments have shown that sesquiterpene lactones have cytotoxicity, potential anticancer activity, and can inhibit platelet-activating factor, achieving an anticoagulant effect.

[0045] Sandalwood extract possesses antibacterial properties and inhibits methionine enzymes, helping to suppress body odor. While sandalwood extract itself doesn't have strong water-holding capacity, it can synergistically enhance the water-holding capacity of other moisturizers. Sandalwood can activate the olfactory receptor OR2AT4 on the surface of keratinocytes, causing calcium ion influx, which further activates Erk1 / 2 and p38MAPK via the cAMP pathway. Long-term use of sandalwood can promote the proliferation and migration of keratinocytes, thereby promoting epidermal healing. Another 2018 study demonstrated that synthetic sandalwood, the substance used in personal care products to provide the sandalwood scent, binds to an olfactory receptor called IGF-1, which regulates the expression and secretion of human hair growth.

[0046] The inventors of this application have discovered that the compound composition of Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract has significantly superior anti-allergic, soothing, and repairing effects compared to the sum of the anti-allergic, soothing, and repairing effects of using Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract alone. Therefore, the composition of Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract has a significant synergistic effect in anti-allergic, soothing, and repairing effects.

[0047] In some embodiments, the anti-inflammatory, soothing, and repairing composition comprises the following components in parts by weight: 10-30 parts of Saposhnikovia divaricata root extract; 10-30 parts of Acer palmatum extract; and 0.01-0.5 parts of sandalwood extract.

[0048] For example, the mass fractions of the Saposhnikovia root extract can be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 parts or any range of two of the above values.

[0049] For example, the mass fractions of the purple bee jelly extract can be 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, or any range of two of the above values.

[0050] For example, the mass fractions of sandalwood extract can be 0.01 parts, 0.03 parts, 0.05 parts, 0.08 parts, 0.1 parts, 0.12 parts, 0.15 parts, 0.18 parts, 0.2 parts, 0.22 parts, 0.25 parts, 0.28 parts, 0.3 parts, 0.32 parts, 0.35 parts, 0.38 parts, 0.4 parts, 0.42 parts, 0.45 parts, 0.48 parts, 0.5 parts, or any range of two of the above values.

[0051] This application verifies through experiments that the combination of the above-mentioned Saposhnikovia root extract, Amaranthus urinaria leaf extract, and Sandalwood extract can significantly inhibit the secretion of LPS-induced inflammatory factors (NO, IL-6) and significantly inhibit the mRNA expression level of UVB-induced inflammatory factors (IL-6, IL-8), and has a synergistic effect, indicating that the anti-inflammatory, soothing, and repairing composition has a good inhibitory effect on inflammation caused by different factors.

[0052] This application also experimentally verified that the combination of Saposhnikovia root extract, Amaranthus urinaria leaf extract, and Sandalwood extract can inhibit the expression of the cold and heat targets TRPV1, TRPV4, and TRPA1. This indicates that the anti-inflammatory, soothing, and repairing composition has a certain alleviating effect on the occurrence of sensitive skin caused by cold and heat.

[0053] Furthermore, according to the experimental results of this application, the combination of Saposhnikovia root extract, Amaranthus urticaria leaf extract, Sandalwood extract, and cyclodextrin not only has a good anti-inflammatory and soothing effect, but also a good barrier repair effect. The combination of Saposhnikovia root extract, Amaranthus urticaria leaf extract, Sandalwood extract, and cyclodextrin can promote the mRNA expression levels of FLG, LOR, ZO-1, and CLND3, proving that the combination has a certain repair effect on the skin barrier.

[0054] Secondly, embodiments of this application provide a method for preparing the anti-inflammatory, soothing, and repairing composition of the first aspect, comprising the following steps: mixing Saposhnikovia root extract, Acer palmatum extract, Sandalwood extract with maltodextrin and water, stirring, to obtain the anti-inflammatory, soothing, and repairing composition.

[0055] In some embodiments of this application, the preparation method of the anti-inflammatory soothing and repairing composition further includes the following step: spray drying the anti-inflammatory soothing and repairing composition to make it into a powder.

[0056] Thirdly, embodiments of this application provide the application of the anti-inflammatory, soothing, and repairing composition of the first aspect in the preparation of cosmetics.

[0057] In some embodiments of this application, cosmetics include anti-inflammatory cosmetics, soothing cosmetics, barrier repair cosmetics, or after-sun repair cosmetics.

[0058] In some embodiments of this application, cosmetics include cosmetics suitable for sensitive skin.

[0059] In some embodiments of this application, cosmetics include at least one of the following: facial mask liquid, lotion, cream, serum, toner, essence, facial cleanser, liquid foundation, foundation cream, concealer, perfume, lotion, makeup remover, blush, eyeshadow, lipstick, and blush.

[0060] The fourth aspect of this application provides a cosmetic composition comprising the anti-inflammatory, soothing, and repairing composition of the first aspect of this invention.

[0061] In some embodiments of this application, the cosmetic composition provided comprises, by weight percentage: 0.1% to 1% anti-inflammatory, soothing, and repairing composition; the remainder being a cosmetic base. Optionally, the cosmetic base includes: 4% glycerin, 2% butylene glycol, 0.5% sodium polyacrylate, 0.05% disodium EDTA, 0.2% potassium cetyl phosphate, 3.2% behenyl alcohol, 1% stearic acid, 3.5% polyglycerol-3 distearate (and) glyceryl stearate citrate, 6% caprylic / capric triglyceride, 0.3% tocopheryl acetate, 2% dicarbonate, 0.5% hydrogenated polydecene, 3% 1,2-pentanediol, 0.3% 1,2-hexanediol, and the remainder being water. The resulting cream has a uniform texture, a delicate feel, is easily absorbed, and has a light sandalwood fragrance, providing a soothing and refreshing sensation.

[0062] This invention proposes the application of a composition containing Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, Santalum album extract, and cyclodextrin in the preparation of soothing and repairing cosmetics, particularly for cosmetics targeting sensitive skin caused by seasonal or temperature changes. The compositions of this invention can be formulated into various dosage forms such as creams, sprays, and powders, and used as active ingredients in cosmetics. The resulting cosmetics can take various forms, including creams, emulsions, masks, toners, lotions, ointments, oils, gels, aerosols, solutions, or film-forming agents.

[0063] Example

[0064] The following embodiments describe the disclosure of this application in more detail. These embodiments are merely illustrative, as various modifications and variations will be apparent to those skilled in the art within the scope of the disclosure of this application. Unless otherwise stated, all parts, percentages, and ratios reported in the following embodiments are based on weight, and all reagents used in the embodiments are commercially available or synthesized by conventional methods and can be used directly without further processing, and the instruments used in the embodiments are commercially available.

[0065] Saposhnikovia divaricata root extract: purchased from Xi'an Xinhe Biotechnology Co., Ltd.;

[0066] PETASITES HYBRIDUS leaf extract: purchased from Xi'an Xinhe Biotechnology Co., Ltd.;

[0067] Sandalwood extract: purchased from Shanghai Yijia Cosmetics Co., Ltd.

[0068] Example 1: Effects of Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, and Santalum album extract on HaCaT cell activity:

[0069] Immortalized human epidermal cells HaCaT cells were used at a rate of 1×10 5 Cells were seeded at a density of 10 μL per well in 96-well plates. After cell adhesion, the original cell culture medium was replaced with 100 μL of DMEM medium containing active ingredients. The concentrations of Saposhnikovia root extract and Amaranthus purpurea leaf extract in the experimental groups were (31.25 ppm, 62.5 ppm, 125 ppm, 250 ppm, 500 ppm, 1000 ppm), respectively; the concentrations of Sandalwood extract in the experimental groups were (0.0078%, 0.015625%, 0.03125%, 0.0625%, 0.125%, 0.25%), respectively. After 24 h, CCK-8 was added to detect cell proliferation activity. 10 μL of CCK-8 solution was added to each well, and the medium was incubated at 37°C for 1 h until it turned orange. The specific incubation time varied depending on the cell type. The absorbance was measured at 450 nm; a higher absorbance indicated greater cell proliferation activity, and vice versa.

[0070] The experimental results are as follows Figures 1 to 3 As shown, Figure 1 A schematic diagram illustrating the effect of different concentrations of Saposhnikovia root extract on cell activity; Figure 2 A schematic diagram illustrating the effect of different concentrations of *Begonia purpurea* leaf extract on cell activity; Figure 3A schematic diagram illustrating the effects of different concentrations of sandalwood extract on cell viability (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). Figures 1 to 3 It can be seen that, compared with the control group, when the concentration of Saposhnikovia root extract is less than or equal to 1000 ppm, the concentration of Acer palmatum leaf extract is less than or equal to 500 ppm, and the concentration of sandalwood extract is less than or equal to 0.0625%, none of them have a toxic effect on the activity of HaCaT cells. Therefore, in subsequent experiments, the concentration of Saposhnikovia root extract and Acer palmatum leaf extract was selected at 500 ppm for experimental research.

[0071] Example 2: Effects of Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, Sandalwood extract, and an anti-inflammatory, soothing, and repairing composition on LPS-induced inflammatory factor secretion:

[0072] RAW264.7 cells were seeded into well plates at a specific concentration and incubated overnight at 37°C in a 5% CO2 incubator. Separate control groups were established, including a blank control group, a model group, a positive control group, a group containing Saposhnikovia root extract, a group containing Amaranthus urinaria leaf extract, a group containing Santalum album extract, and a group containing an anti-inflammatory, soothing, and repairing compound. Drug administration was initiated when the cell deposition rate in the well plates reached 50-60%. The blank control group received culture medium without LPS inducer, the LPS model group received culture medium containing LPS inducer, the positive control group received culture medium containing LPS inducer and the positive control dexamethasone, and the sample groups received culture medium containing LPS inducer and the concentrations of samples shown in Table 1.

[0073] Table 1. Effects of LPS-induced inflammatory factor secretion on experimental sample groups

[0074]

[0075] After drug administration, the well plate was placed in an incubator (37℃, 5% CO2) for culture; cell morphology was observed and photographed under a microscope; after cell incubation, the cell supernatant was collected and the contents of inflammatory factors NO and IL-6 were measured using the corresponding ELISA reagents.

[0076] Figure 4 The experimental results show the effects of Saposhnikovia root extract, Amaranthus purpurea leaf extract, Sandalwood extract, and an anti-inflammatory, soothing, and repairing composition on LPS-induced NO secretion. Figure 5 Figure 1 shows the experimental results (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001) of the effects of Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, Santalum album extract, and an anti-inflammatory, soothing, and repairing combination on LPS-induced IL-6 secretion. Figure 4 and Figure 5It was found that the anti-inflammatory and soothing repair composition containing Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract significantly inhibited the secretion of NO and IL-6 by LPS-induced macrophages, and was significantly superior to the combined effects of the individual extracts of Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract. The anti-inflammatory and soothing repair composition containing Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract achieved significant anti-inflammatory efficacy at a low dosage.

[0077] Example 3: Effects of Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, Sandalwood extract, and an anti-inflammatory, soothing, and repairing composition on UVB-induced secretion of inflammatory factors:

[0078] Cell culture: Immortalized human keratinocytes were cultured in a medium containing 10% fetal bovine serum and 1% penicillin-dextrose antibody (1×10⁻⁶). 5 Cells were cultured in DMEM medium containing 100 mg / L penicillin and 100 mg / L streptomycin at 37°C and 5% CO2 until confluence reached 85-95%. Logarithmic growth phase cells were digested with 0.05% trypsin, and the digestion was terminated with DMEM medium containing 10% serum. Cells were counted using a cell counting chamber, and appropriate cells were seeded into 6-well plates and incubated at 37°C and 5% CO2 until confluence reached 60-70%. The original culture medium in the 6-well plates was replaced with serum-free medium, and after starvation for two hours, the root extract of *Saposhnikovia divaricata*, leaf extract of *Acer palmatum*, sandalwood extract, and the anti-inflammatory, soothing, and repairing composition of this invention were added. After treatment for 4 hours, the medium was replaced with PBS, and the solution was added at 30 mJ / cm³. 2 Keratinocytes were irradiated with UVB and cultured for 24 hours. Trizol was then added, and the cells were collected. A blank control group and a model group (treated with 30 mJ / cm²) were established. 2 UVB irradiation of keratinocytes (without sample added).

[0079] Table 2. Effects of UVB-induced secretion of inflammatory factors on experimental sample groups.

[0080]

[0081] RNA Extraction: Cell Collection (on Ice): Wash cells cultured in 6-well plates twice with pre-chilled sterile PBS. Add 500 μl of TRIzol to each well and gently pipette to collect cell lysates into 1.5 ml EP tubes. Centrifuge at 12000 g for 10 min in a pre-chilled 4°C high-speed centrifuge. Transfer the supernatant to a new RNase-free 1.5 ml EP tube and discard the tissue pellet. Add 200 μl of chloroform to the supernatant and vortex 15 times to thoroughly mix. Keep the tube cap closed and incubate at room temperature for 5 min to allow for natural phase separation. Centrifuge at 12000 g for 15 min in a 4°C high-speed centrifuge. Carefully aspirate the upper aqueous phase into a new RNase-free 1.5 ml EP tube, taking special care not to touch the central pellet to avoid protein contamination. Add 500 μl of isopropanol to the supernatant and gently vortex. Keep the tube cap closed and incubate at room temperature for 5 min to allow for complete mRNA extraction. Centrifuge at 12000g for 10 min at 4℃, discard the supernatant, and add 75% ethanol solution prepared with DEPC water. Gently invert to wash the mRNA precipitate. Centrifuge at 7500g for 5 min at 4℃, discard the supernatant, and wash once more with 75% ethanol solution. Centrifuge at 7500g for 5 min at 4℃, discard the supernatant, and aspirate as much residual liquid as possible. Let stand at room temperature with the cap open for 30-60 min to allow residual ethanol to evaporate. Add an appropriate amount of DEPC water to dissolve the mRNA, determine the concentration, and perform reverse transcription according to the reverse transcription kit instructions.

[0082] Reverse transcription PCR: 20 μl system: 10× buffer 2 μl, 2.5 mM dNTP 2 μl, DNA polymerase (Taq) 0.5 μl, primers 2 μl, template (cDNA obtained from reverse transcription diluted 10-fold as template) 2-10 μl, ultrapure water to bring the system to 20 μl. Reaction conditions: Pre-denaturation 94℃ for 5 min; 30 cycles: denaturation 94℃ for 30 s, annealing 60℃ for 30 s, extension 72℃ for 30 s; final extension 72℃ for 10 min.

[0083] Quantitative Real-Time PCR: System (20 μl): 10 μl of 2×SYBR, 2-5 μl of template (cDNA obtained from reverse transcription diluted 10-fold as template), 1-2 μl of primers, and ultrapure water to bring the system to 20 μl. Reaction conditions: Pre-denaturation 94℃ for 5 min; 40 cycles: denaturation 94℃ for 30 s, annealing 60℃ for 30 s, extension 72℃ for 30 s (real-time fluorescence imaging); melting curve 94℃ for 30 s, 60℃ for 30 s, 72℃ for 1 s (real-time fluorescence imaging during the heating process).

[0084] Figure 6 Figure 1 shows the experimental results of the effects of Saposhnikovia divaricata root extract, Amaranthus purpurea leaf extract, Santalum album extract, and an anti-inflammatory, soothing, and repairing composition on UVB-induced IL-6 secretion. Figure 7Figure 1 shows the experimental results (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001) of the effects of Saposhnikovia divaricata root extract, Amaranthus urinaria leaf extract, Santalum album extract, and an anti-inflammatory, soothing, and repairing composition on UVB-induced IL-8 secretion. Figure 6 and Figure 7 It was found that the anti-inflammatory and soothing repair composition containing Saposhnikovia root extract, Amaranthus praecox leaf extract, and Sandalwood extract significantly inhibited the mRNA expression of IL-6 and IL-8 in UVB-induced keratinocytes (HaCaT), and was superior to the combined effect of each extract alone. Furthermore, the anti-inflammatory and soothing repair composition containing these extracts achieved significant anti-inflammatory efficacy at relatively low dosages.

[0085] Example 4: Effect of anti-inflammatory, soothing, and repairing composition on TRPA1 expression induced by cold stimulation:

[0086] The keratinocyte culture procedure involves in vitro culture of keratinocytes on cell culture plates under constant temperature and humidity conditions. When the cell fusion rate reaches 70-80%, the treated cells are digested, then uniformly counted by pipetting, and the cell density is adjusted to 1-2 mL per well. The cells are then inoculated into 6-well plates for further culture. The sample addition procedure involves diluting and adding the analyte when the cell fusion rate reaches 70-80%. Serum-free medium is used as the diluent, and the composition is added to the cells. After incubation for 4 hours, 100 μL of room temperature PBS is added to the control group, while PBS at 4 degrees Celsius is added to the model group and sample group. Treatment lasts for 30 minutes to obtain the sample group and cold-stimulated model group, respectively. After the cold stimulation treatment, the PBS is removed, and fresh serum-free culture medium is added to the wells. The cells are then cultured for another 24 hours. After 24 hours, cells are collected, RNA is extracted, and the expression level of TRPA1 mRNA in HaCaT is detected using qPCR. Figure 8 The experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of TRPA1 mRNA induced by cold stimulation are shown in the figure (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). As shown in the figure, incubation in a PBS ice bath at 4°C significantly activated the expression level of TRPA1 mRNA in HaCaT cells. Treatment with the composition significantly reduced the increase in TRPA1 mRNA levels induced by cold stimulation, demonstrating that the composition has a significant inhibitory effect on the expression level of TRPA1 mRNA induced by cold.

[0087] Example 5: Experiment on the effect of anti-inflammatory, soothing, and repairing composition on TRPV1 mRNA expression levels induced by heat stimulation:

[0088] The keratinocyte culture procedure involves culturing keratinocytes in vitro on cell culture plates under constant temperature and humidity conditions. When the cell fusion rate reaches 70-80%, the treated cells are digested, then uniformly counted by pipetting, and the cell density is adjusted to 1-2 mL per well. The cells are then added to a 6-well plate for further culture. The sample delivery procedure involves diluting and adding the analyte when the cell fusion rate reaches 70-80%. Serum-free medium is used as the diluent, and the mixture is added to the cells. The cells are then incubated at 37°C for 24 hours. After incubation, the cells are collected, RNA is extracted, and the expression level of TRPV1 mRNA in HaCaT is detected using qPCR. Figure 9 The experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of TRPV1 mRNA induced by heat stimulation are shown in the figure (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). As can be seen from the figure, treatment with the composition significantly reduced the increase in TRPV1 mRNA levels, demonstrating that the composition has a significant inhibitory effect on the expression of TRPV1 mRNA.

[0089] Example 6: Effect of anti-inflammatory, soothing, and repairing composition on TRPV4 mRNA expression levels induced by UVB stimulation

[0090] The keratinocyte culture procedure involves in vitro culture of keratinocytes on cell culture plates under constant temperature and humidity conditions. When the cell fusion rate reaches 70-80%, the treated cells are digested, then uniformly counted by pipetting, and the cell density is adjusted to 1-2 mL per well. The cells are then inoculated into 6-well plates for culture. The sample delivery procedure involves diluting and adding the analyte when the cell fusion rate reaches 70-80%. Serum-free medium is used as the diluent, and the composition is added to the cells. After incubation for 4 hours, PBS is replaced. The control group is covered with aluminum foil, while the sample and model groups are irradiated with UVB at 30 mJ / cm². After irradiation, the PBS is removed, and fresh serum-free culture medium is added to the wells. Culture is continued for 24 hours. After 24 hours, cells are collected, RNA is extracted, and the expression level of TRPV4 mRNA in HaCaT is detected using qPCR.

[0091] Figure 10The experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of TRPV4 mRNA induced by UVB stimulation are shown in the figure (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). As shown in the figure, UVB irradiation can significantly activate the expression level of TRPV4 mRNA in HaCaT cells. Treatment with the composition significantly reduces the increase in TRPV4 mRNA level induced by UVB, demonstrating that the composition has a significant inhibitory effect on the expression level of TRPV4 mRNA induced by UVB.

[0092] Example 7: Effect of anti-inflammatory, soothing, and repairing composition on gene expression levels of skin barrier-related proteins induced by UVB.

[0093] The keratinocyte culture procedure involves in vitro culture of keratinocytes on cell culture plates under constant temperature and humidity conditions. When the cell fusion rate reaches 70-80%, the treated cells are digested, then uniformly counted by pipetting, and the cell density is adjusted to 1-2 mL per well. The cells are then inoculated into 6-well plates for culture. The sample delivery procedure involves diluting and adding the analyte when the cell fusion rate reaches 70-80%. Serum-free medium is used as the diluent, and the composition is added to the cells. After incubation for 4 hours, PBS is replaced. The control group is covered with aluminum foil, while the sample and model groups are irradiated with UVB at 30 mJ / cm². After irradiation, the PBS is removed, and fresh serum-free culture medium is added to the wells. Culture is continued for 24 hours. After 24 hours, cells are collected, RNA is extracted, and the expression levels of FLG and LOR mRNA in HaCaT are detected using qPCR. Figure 11 Figure showing the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of FLG mRNA induced by UVB.

[0094] Figure 12 The experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on LOR mRNA expression levels induced by UVB are shown in the figure (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). As shown in the figure, UVB irradiation significantly inhibited the expression of FLG and LOR mRNA levels in HaCaT cells. Treatment with the composition significantly promoted the increase of FLG and LOR mRNA levels, demonstrating that the composition has a protective effect against UVB-induced skin barrier damage and can repair the skin barrier.

[0095] Example 8: Effect of the composition on UVB-induced expression levels of tight junction-related genes

[0096] Keratinocyte culture steps: Keratinocytes are cultured in vitro on cell culture plates under constant temperature and humidity conditions. When the cell fusion rate reaches 70-80%, the treated cells are digested, then uniformly counted by pipetting, and the cell density is adjusted to 1-2 mL per well. The cells are then inoculated into 6-well plates for culture. Sample delivery steps: When the cell fusion rate reaches 70-80%, the analyte is diluted and added. Serum-free medium is used as the diluent. The composition is added to the cells, and after incubation for 4 hours, the solution is replaced with PBS. The control group is covered with aluminum foil, while the sample group and model group are treated with 30 mJ / cm² PBS. 2 The cells were irradiated with UVB. After irradiation, PBS was removed, and fresh serum-free culture medium was added to the wells. The cells were then cultured for another 24 hours. After 24 hours, the cells were collected, RNA was extracted, and the expression levels of FLG and LOR mRNA in HaCaT cells were detected by qPCR. Figure 13 Figure showing the experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of ZO-1 mRNA induced by UVB; Figure 14 The experimental results of the effect of the anti-inflammatory, soothing, and repairing composition on the expression level of CLND3 mRNA induced by UVB are shown in the figure (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). As shown in the figure, UVB irradiation significantly inhibited the expression levels of ZO-1 and CLND3 mRNA in HaCaT cells. Treatment with the composition significantly promoted the increase of ZO-1 and CLND3 mRNA levels, demonstrating that the composition has a certain protective effect against UVB-induced skin barrier damage and can repair the skin barrier.

[0097] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and not to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application, and they should all be covered within the scope of the claims and specification of this application. In particular, as long as there is no structural conflict, the various technical features mentioned in the embodiments can be combined in any way. This application is not limited to the specific embodiments disclosed herein, but includes all technical solutions falling within the scope of the claims.

Claims

1. An anti-inflammatory, soothing, and repairing composition, characterized in that, By weight, it contains: 20 parts of Saposhnikovia root extract; 20 portions of purple bee jelly extract; Sandalwood extract 0.05 parts.

2. The method for preparing the anti-inflammatory, soothing, and repairing composition according to claim 1, characterized in that, The process includes the following steps: mixing the Saposhnikovia root extract, the Amaranthus urinaria extract, the sandalwood extract with maltodextrin and water, and stirring to obtain the anti-inflammatory, soothing, and repairing composition.

3. The method for preparing the anti-inflammatory, soothing, and repairing composition according to claim 2, characterized in that, It also includes the following step: spray drying the anti-inflammatory, soothing, and repairing composition to form a powder.

4. The use of the anti-inflammatory, soothing, and repairing composition according to claim 1 in the preparation of cosmetics.

5. The application according to claim 4, characterized in that, The cosmetics mentioned include anti-inflammatory cosmetics, soothing cosmetics, barrier repair cosmetics, or after-sun repair cosmetics.

6. The application according to claim 4, characterized in that, The cosmetics mentioned include those suitable for sensitive skin.

7. The application according to claim 4, characterized in that, The cosmetics include at least one of lotions and creams.

8. The application according to claim 4, characterized in that, The cosmetics include at least one of the following: facial mask liquid, serum, toner, essence, facial cleanser, liquid foundation, foundation cream, concealer, perfume, lotion, makeup remover, blush, eyeshadow, lipstick, and blush.

9. A cosmetic product, characterized in that, Includes the anti-inflammatory, soothing, and repairing composition as described in claim 1.

10. The cosmetic product according to claim 9, characterized in that, In terms of mass percentage, it includes: The anti-inflammatory, soothing, and repairing composition: 0.1% to 1%; The remainder is cosmetic base.

Images