An anti-inflammatory soothing traditional Chinese medicine compound ethyl acetate extract, and a preparation method and application thereof

Abstract

This invention discloses an anti-inflammatory and soothing compound herbal extract in ethyl acetate, its preparation method, and its application. The compound herbal formula consists of 7-20 parts by weight of Sophora japonica flower, 7-20 parts by weight of lotus leaf, and 2-6 parts by weight of licorice root. The three herbs work synergistically: Sophora japonica flower acts as the principal herb for cooling the blood, lotus leaf acts as the secondary herb for promoting diuresis and clearing heat, and licorice root acts as the adjuvant herb for detoxification and harmonizing the middle jiao. This combination achieves the effects of cooling the blood without causing stagnation, and removing dampness without causing excessive dryness, thus exerting a synergistic effect of cooling the blood and reducing swelling, clearing heat and removing dampness, and soothing and calming the nerves. This invention extracts the compound herbal formula with 70% ethanol, followed by liquid-liquid extraction with ethyl acetate to obtain an extract rich in flavonoids and alkaloids. Functional experiments show that the anti-inflammatory and soothing effect of this extract is significantly better than that of the unextracted ethanol extract. This invention also prepares the extract into a gel with a refreshing feel and good transdermal penetration, and confirms its efficacy in improving skin inflammation when applied topically, providing a safe and effective solution for the care of sensitive skin.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine compound, its preparation method and application, and more particularly to an anti-inflammatory and soothing traditional Chinese medicine compound ethyl acetate extract, its preparation method and application. Background Technology

[0002] The fast pace of modern life and high-intensity work patterns constantly expose the skin to multiple internal and external stressors. External environmental factors such as ultraviolet radiation, fine particulate matter, and blue light from electronic screens can induce oxidative stress and chronic micro-inflammation in the skin, damaging the integrity of the epidermal barrier. Internally, factors such as mental stress, sleep deprivation, and dietary imbalances can easily lead to abnormal levels of inflammatory mediators such as cortisol, resulting in sensitive and inflammatory reactions such as redness, burning, itching, peeling, and acne. Prolonged mask-wearing for protection creates a warm, humid, and enclosed microenvironment on the face, further exacerbating skin discomfort and significantly increasing the proportion of people with sensitive skin. Against this backdrop, developing products that can safely and effectively inhibit inflammatory responses such as redness, swelling, heat, and pain, and provide immediate soothing and repair, has become an urgent need in the field of skin care.

[0003] Currently available anti-inflammatory and soothing products mostly rely on chemically synthesized active ingredients, such as corticosteroids, antihistamines, or nonsteroidal anti-inflammatory drugs (NSAIDs). While these products are fast-acting, long-term use can easily lead to side effects such as skin atrophy, telangiectasia, pigmentation, and even hormone-dependent dermatitis, raising growing concerns about their safety. Consumers are increasingly favoring natural herbal extracts that are derived from natural sources, are gentle, non-addictive, and have proven efficacy, hoping to find new anti-inflammatory and soothing solutions from traditional Chinese medicine and other natural resources. This points to an important direction for the development of green and safe skin care products.

[0004] Traditional Chinese medicine (TCM) has a unique and systematic understanding of the pathogenesis and treatment of skin inflammation. TCM believes that although skin diseases manifest externally, their root cause often lies internally, with the core pathogenesis revolving around "heat," "toxins," "dampness," and "wind." Based on this, it establishes the treatment principles of clearing heat and cooling the blood, dispelling dampness and detoxifying, and relieving wind and itching. External application of TCM, through multi-component regulation of NF-κB and MAPK signaling pathways, synergistically inhibits the release of pro-inflammatory mediators such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), reduces capillary permeability, and promotes the expression of barrier-related proteins, achieving multi-target anti-inflammatory and repair effects. It also boasts the advantages of safety and gentleness, highly meeting the modern skincare demand for long-lasting soothing effects. However, traditional TCM external dosage forms, such as powders and extracts, generally suffer from significant loss of active ingredients, easy oxidation and deterioration, and poor transdermal absorption; ordinary creams, on the other hand, have a greasy and heavy feel, easily clogging pores and resulting in low user compliance.

[0005] In summary, existing skin anti-inflammatory and soothing products suffer from problems such as significant side effects from chemical ingredients, weak efficacy due to the mixed composition of crude Chinese herbal extracts, and poor skin feel due to their formulations. Summary of the Invention

[0006] Purpose of the invention: The purpose of this invention is to provide an anti-inflammatory and soothing compound ethyl acetate extract of traditional Chinese medicine, its preparation method, and its application.

[0007] Technical solution: The anti-inflammatory and soothing compound ethyl acetate extract of traditional Chinese medicine is prepared from Sophora japonica flower, lotus leaf and licorice.

[0008] The ethyl acetate extract of the traditional Chinese medicine compound is preferably prepared from 7-20 parts by weight of Sophora japonica flowers, 7-20 parts by weight of lotus leaves, and 2-6 parts by weight of licorice. Most preferably, it is prepared from 10 parts by weight of Sophora japonica flowers, 10 parts by weight of lotus leaves, and 3 parts by weight of licorice.

[0009] The traditional Chinese medicine compound formula described in this invention follows the following traditional Chinese medicine theories:

[0010] Traditional Chinese medicine (TCM) has a profound understanding of the pathogenesis of skin inflammation, often basing its theories on "heat," "toxin," "dampness," and "wind." The *Suwen* (Plain Questions) states, "All pain, itching, and sores belong to the heart," meaning the heart governs blood vessels. When pathogenic heat enters the blood, it causes blood heat to stagnate, manifesting externally as redness, swelling, and burning sensations on the skin. It also states, "All dampness and swelling belong to the spleen," meaning that when the spleen fails to function properly, damp heat rises, leading to oily skin, papules, and itching. Therefore, external treatment focuses on clearing heat and cooling the blood, dispelling dampness and reducing swelling, and soothing and dispersing. The TCM compound formula described in this invention is based on this principle and is formulated accordingly. Sophora japonica flowers are slightly cold in nature and bitter in taste. They enter the liver and large intestine meridians, effectively cooling the blood and stopping bleeding, clearing liver heat, and directly clearing heat from the blood. They are particularly suitable for skin redness and burning caused by blood heat. Lotus leaves are neutral in nature and bitter in taste. They are light, clear, and dispersing, clearing summer heat and promoting diuresis, and promoting the upward movement of clear yang. They can expel damp-heat from the skin surface, allowing the pathogens to have an outlet and helping to reduce redness and swelling. Licorice is neutral in nature and sweet in taste. It clears heat and detoxifies, relieves spasms and pain, and can harmonize the other herbs. It not only enhances the effect of cooling the blood and reducing swelling, but also makes the bitter and cold herbs of the whole formula sweet and mild, without harming the skin. The combination of the three herbs, with Sophora japonica flowers cooling the blood as the chief herb, lotus leaves promoting diuresis and clearing heat as the assistant herb, and licorice detoxifying and harmonizing as the adjuvant herb, cools the blood without causing stagnation and removes dampness without causing dryness. Together, they achieve the effects of cooling the blood and reducing swelling, clearing heat and removing dampness, and soothing the skin. The theory is consistent with the inflammatory manifestations of sensitive skin such as redness, swelling, heat, and pain. Modern research has also confirmed that flavonoids such as rutin contained in Sophora japonica flowers can inhibit inflammatory factors and hyaluronidase, and reduce capillary permeability; alkaloids such as lotus leaf alkaloids in lotus leaves have antibacterial and anti-inflammatory activities; glycyrrhizic acid in licorice has steroid-like anti-inflammatory effects, which can significantly relieve redness, swelling and itching. The three drugs work together to exert anti-inflammatory and soothing effects on multiple targets.

[0011] The present invention also provides a method for preparing the ethyl acetate extract of the traditional Chinese medicine compound, wherein the extract is first extracted with ethanol and then liquid-liquid extracted with ethyl acetate.

[0012] This method utilizes modern extraction and separation techniques to enrich flavonoids and alkaloids with anti-inflammatory and soothing activities in traditional Chinese medicine compound extracts, which helps to improve the quality and application value of external Chinese medicine products.

[0013] The preferred preparation method includes the following steps: taking Sophora japonica flowers, lotus leaves and licorice, adding ethanol, heating and refluxing to extract, and obtaining a filtrate; concentrating the obtained filtrate under reduced pressure, and then freeze-drying to obtain a dry extract; suspending the dry extract in water evenly, adding ethyl acetate for extraction, and obtaining an extract; concentrating the obtained extract under reduced pressure, and then freeze-drying to obtain an extract.

[0014] The ethanol is preferably 60-80% ethanol. Most preferably, it is 70% ethanol.

[0015] The preferred preparation method includes the following steps: Sophora japonica flowers, lotus leaves, and licorice root are mixed in a ratio of 10:10:3. Approximately 10 parts by weight of each ingredient are added to 10 times the volume of 70% ethanol. The mixture is heated under reflux and extracted twice, 1 hour each time. The filtrates are combined, concentrated under reduced pressure to recover the solvent, and then freeze-dried to obtain a dry extract. The dry extract is then suspended in an appropriate amount of water and extracted three times with the same volume of ethyl acetate. The extracts are combined, concentrated under reduced pressure to recover the solvent, freeze-dried, and then ground into a fine powder.

[0016] This invention also provides the application of the ethyl acetate extract of the traditional Chinese medicine compound in the preparation of topical formulations with anti-inflammatory and soothing effects. The ethyl acetate extract of the traditional Chinese medicine compound has anti-inflammatory and soothing effects and can be used as a cosmetic ingredient.

[0017] This invention also provides a gel with anti-inflammatory and soothing effects, comprising the ethyl acetate extract of the aforementioned traditional Chinese medicine compound and matrix excipients. The gel has characteristics such as high water content, good spreadability, a refreshing feel, and the ability to form a breathable protective film, making it an ideal carrier for immediate skin soothing.

[0018] The gelling agent is preferably made from the following raw materials in weight percentages: 1-5% ethyl acetate extract of traditional Chinese medicine compound, 0.6-1% carbomer 940, 0.9-1.2% triethanolamine, 0.1-0.14% low molecular weight sodium hyaluronate, 10-14% glycerol, 1-3% azone, and 2-4% Tween 80, with the balance being water. The water is preferably ultrapure water.

[0019] The present invention also provides a method for preparing the gelling agent, comprising the following steps:

[0020] (1) Dissolve low molecular weight sodium hyaluronate in glycerol and mix well to obtain a compound moisturizer; dissolve the ethyl acetate extract of the traditional Chinese medicine compound in Tween 80, then add the compound moisturizer and water, dissolve and then add azone, mix well to form phase A;

[0021] (2) Add Carbomer 940 to the remaining compound humectant, then add water and stir to make Carbomer 940 swell and disperse, then stir evenly to form phase B;

[0022] (3) Add phase B to phase A, stir evenly, then add triethanolamine and stir until gel-like to obtain the gelling agent.

[0023] The preferred method for preparing the gelling agent includes the following steps:

[0024] (1) Dissolve 0.1-0.14% low molecular weight sodium hyaluronate in 10-14% glycerol and mix well to obtain a compound moisturizer (low molecular weight sodium hyaluronate: glycerol = 1: 100). Dissolve 1-5% of the compound Chinese medicine ethyl acetate extract in 2-4% Tween 80 in a beaker, then add appropriate amounts of compound moisturizer and ultrapure water respectively. After dissolving, add 1-3% azone and mix well to form phase A.

[0025] (2) Add 0.6~1% Carbomer 940 to the remaining compound humectant, then add an appropriate amount of ultrapure water, stir to make Carbomer 940 swell and disperse (20~30 min), then stir evenly to form phase B;

[0026] (3) Add phase B to phase A, stir evenly, then add 0.9~1.2% triethanolamine, and stir continuously until gel form is obtained, thus obtaining the gel formulation product.

[0027] The present invention also provides the use of the gel in improving skin inflammation.

[0028] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0029] 1. The ethyl acetate extract of the traditional Chinese medicine compound (YSYZ) has significant anti-inflammatory effects: This invention uses Tg (mpx:EGFP) transgenic zebrafish with neutrophil green fluorescent labeling to objectively evaluate the inhibitory effect of the test sample on the inflammatory response induced by sodium dodecyl sulfate (SLS) in zebrafish. It was found that YSYZ can inhibit the migration of neutrophils to the inflammatory site in a dose-dependent manner, and its effect is better than that of 70% ethanol extract (HHG).

[0030] 2. The ethyl acetate extract of the traditional Chinese medicine compound can inhibit abnormal angiogenesis: Skin redness is often accompanied by chronic inflammation, which stimulates the release of large amounts of pro-angiogenic factors such as vascular endothelial growth factor, further promoting angiogenesis; newly formed blood vessels are usually more permeable, which in turn aggravates inflammation and redness, forming a vicious cycle of "inflammation-angiogenesis-intensified inflammation"; this invention uses Tg (fli1:EGFP) transgenic zebrafish labeled with vascular green fluorescence and found that YSYZ can significantly inhibit the formation of intersegmental vessels (ISVs) in juvenile zebrafish, and the inhibitory effect of YSYZ is better than that of HHG; suggesting that in addition to direct anti-inflammatory effects, YSYZ can also fundamentally alleviate the symptoms of skin redness and flushing by improving abnormal angiogenesis caused by inflammatory stimulation;

[0031] 3. The ethyl acetate extract of the traditional Chinese medicine compound has a nerve-soothing effect: Skin inflammation can cause discomfort such as itching and stinging, leading to unconscious scratching behavior; This invention uses a zebrafish behavioral analysis system to objectively quantify the effect of the test sample on the movement distance of zebrafish juveniles after SLS stimulation, and found that YSYZ can dose-dependently slow down the movement of juvenile fish, and its soothing effect is better than HHG.

[0032] 4. Topical application of the YSYZ gel can improve skin inflammation: This invention uses an imiquimod (IMQ)-induced mouse psoriasis-like dermatitis model to evaluate the effects of topical application of 4% HHG gel and 4% YSYZ gel on inflammatory infiltration and angiogenesis in skin lesions. It was found that the anti-inflammatory and anti-angiogenic effects of 4% YSYZ gel were significantly better than those of 4% HHG gel.

[0033] 5. The YSYZ gel preparation process is stable and controllable: The raw materials used in this invention are all Chinese medicinal herbs that are both food and medicine. The separation technology of "70% ethanol extraction-ethyl acetate enrichment" can effectively enrich active components such as flavonoids and alkaloids. Through multi-factor orthogonal experimental design, using carbomer 940, glycerol and triethanolamine as the factors of consideration, and using appearance, spreadability, viscosity, centrifugal stability and heat and cold resistance as comprehensive scoring indicators, the optimal preparation process of the compound Chinese medicine ethyl acetate extract gel was selected. The prepared gel has a uniform appearance, good spreadability and good stability, which meets the needs of modern consumers, especially people with sensitive skin, for high-quality skin care products and has broad application prospects. Attached Figure Description

[0034] Figure 1 The total ion chromatograms for qualitative analysis of chemical components using UPLC-ESI-Q TRAP-MS / MS are shown in Figure A (70% ethanol extract of a traditional Chinese medicine compound prepared from Sophora japonica, lotus leaf, and licorice (HHG); Figure B (ethyl acetate extract of the 70% ethanol extract of the traditional Chinese medicine compound (YSYZ); positive and negative ion modes).

[0035] Figure 2 To evaluate the effects of HHG and YSYZ on neutrophil migration in a zebrafish inflammation model induced by sodium dodecyl sulfate (SLS) using Tg (mpx:EGFP) transgenic zebrafish (Ctrl group: normal control group; SLS group: SLS-induced zebrafish inflammation model group; HHG group: 50, 100, 200 μg / mL 70% ethanol extract of traditional Chinese medicine compound; YSYZ group: 50, 100, 200 μg / mL ethyl acetate extract of traditional Chinese medicine compound);

[0036] Figure 3 To evaluate the effects of HHG and YSYZ on angiogenesis in zebrafish larvae using Tg(fli1:EGFP) transgenic zebrafish (Ctrl group: normal control group; SLS group: inflammation model group; VRI group: positive control group for vascular endothelial growth factor receptor tyrosine kinase inhibitor II (VRI) inhibiting angiogenesis in zebrafish larvae; HHG group: 50, 100, 200 μg / mL 70% ethanol extract of traditional Chinese medicine compound; YSYZ group: 50, 100, 200 μg / mL ethyl acetate extract of traditional Chinese medicine compound);

[0037] Figure 4 To evaluate the effects of HHG and YSYZ on the movement distance of zebrafish juveniles after SLS stimulation using a zebrafish behavioral analysis system (Ctrl group: normal control group; SLS group: inflammation model group; HHG group: 50, 100, 200 μg / mL 70% ethanol extract of traditional Chinese medicine compound; YSYZ group: 50, 100, 200 μg / mL ethyl acetate extract of traditional Chinese medicine compound; data are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons among multiple groups, and Dunnett's multiple comparisons were used for pairwise comparisons; compared with the Ctrl group, ### P<0.001; compared with the SLS group, ** P<0.01, *** P<0.001);

[0038] Figure 5 To evaluate the effects of topical 4% HHG gel and 4% YSYZ gel on skin lesions using an imiquimod (IMQ)-induced mouse psoriasis-like dermatitis model (A shows the skin lesions on the back of mice in each group (n=8); B shows the HE staining results of the skin lesions on the back of mice in each group (n=6)). Detailed Implementation

[0039] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0040] Example 1

[0041] This embodiment provides an anti-inflammatory and soothing compound ethyl acetate extract of traditional Chinese medicine and its preparation method, and performs qualitative analysis on the chemical composition of the final product (ethyl acetate extract (YSYZ)) and the intermediate product (70% ethanol extract of compound traditional Chinese medicine (HHG)) based on UPLC-ESI-Q TRAP-MS / MS.

[0042] 1. Sample preparation

[0043] According to the ratio of Sophora japonica flower: lotus leaf: licorice root = 10:10:3, take 100 g of Sophora japonica flower, 100 g of lotus leaf, and 30 g of licorice root, add 10 times the amount of 70% ethanol, heat and reflux twice, 1 h each time, combine the two filtrates, concentrate under reduced pressure to recover the solvent, and freeze-dry to obtain a dry extract. The yield is 22.47%. Weigh the dry extract of 70% ethanol equivalent to 1 g of crude drug, dissolve it in 70% methanol, filter through a 0.22 μm microporous membrane to obtain the intermediate product test solution.

[0044] The 70% ethanol extract of the above-mentioned traditional Chinese medicine compound was taken, suspended evenly in an appropriate amount of water, and then extracted three times with the same volume of ethyl acetate. The extracts were combined, concentrated under reduced pressure to recover the solvent, and freeze-dried to obtain the ethyl acetate extract. The yield was 2.11%. Ethyl acetate extract equivalent to 1 g of crude drug was weighed, dissolved in 80% methanol, and filtered through a 0.22 μm microporous membrane to obtain the final product test solution.

[0045] 2. Chromatographic conditions

[0046] (1) Chromatographic column: Agilent SB-C 18 1.8 µm, 2.1 mm × 100 mm; (2) Mobile phase: Phase A is ultrapure water (with 0.1% formic acid added), and Phase B is acetonitrile (with 0.1% formic acid added); (3) Elution gradient: The proportion of Phase B is 5% at 0.00 min, and the proportion of Phase B increases linearly to 95% within 12.00 min and is maintained at 95% for 2 min. The proportion of Phase B decreases to 5% within 14.00-24.00 min; (4) Flow rate 0.35 mL / min, column temperature 40℃, and injection volume 2 μL.

[0047] 3. Mass spectrometry conditions

[0048] Electrospray ionization (ESI) temperature was 500 °C; ion spray voltage (IS) was 5500 V (positive ion mode) / -4500 V (negative ion mode); ion source gas I (GSI), gas II (GSII), and curtain gas (CUR) were set to 50, 60, and 25 psi, respectively, and collision-induced ionization parameters were set to high. QQQ scans used MRM mode with the collision gas (nitrogen) set to medium. DP and CE for each MRM ion pair were optimized through further declustering potential (DP) and collision energy (CE). A specific set of MRM ion pairs was monitored at each epoch based on the eluted components.

[0049] The substance was qualitatively identified based on secondary spectral information. Isotope signals were removed during the analysis. (K content is not specified.) + Ions, Na + Ions, NH4 + The repetitive signals of ions, as well as the repetitive signals of fragment ions that are themselves other substances with larger molecular weights, were analyzed. Metabolite quantification was performed using multiple reaction monitoring (MRM) mode of triple quadrupole mass spectrometry.

[0050] 4. Analysis Results

[0051] like Figure 1 As shown, compared with the 70% ethanol extract (HHG), the ethyl acetate extract (YSYZ) showed a significant enrichment of flavonoids and alkaloids within a retention time of 8–14 min, and these two types of components have good anti-inflammatory and soothing effects.

[0052] Example 2

[0053] This example evaluates the effects of HHG and YSYZ on neutrophil migration in a zebrafish inflammation model induced by sodium dodecyl sulfate (SLS) in Example 1, based on Tg (mpx:EGFP) transgenic zebrafish.

[0054] 1. Effects of HHG and YSYZ on neutrophil migration in a SLS-induced zebrafish inflammation model

[0055] Preparation of test solution: Weigh appropriate amounts of SLS, 70% ethanol extract (HHG), and ethyl acetate extract (YSYZ) used in the experiment, dissolve them in an appropriate amount of DMSO, dilute with zebrafish embryo culture medium, and bring to a suitable volume to obtain the test solution. Before use, take an appropriate amount of the stock solution and dilute it with culture medium to the required concentration.

[0056] Tg (mpx:EGFP) transgenic zebrafish larvae in good condition and developed to 3 days (3 dpf) were selected and placed in 24-well plates, with 10 larvae per well as one group. The Ctrl group was given zebrafish embryo culture medium containing 0.1% DMSO, the SLS group was given 50 μg / mL SLS, the HHG group was given 50 μg / mL SLS and 50, 100, and 200 μg / mL HHG simultaneously, and the YSYZ group was given 50 μg / mL SLS and 50, 100, and 200 μg / mL YSYZ simultaneously. After incubation at 28℃ for 24 h, the zebrafish larvae in each group were washed with embryo culture medium and anesthetized with 0.02% tricaine. The larvae were then placed on biconcave slides, kept in a lateral recumbent position, and their neutrophils were observed using an upright fluorescence microscope. Meanwhile, Image-Pro Plus 6.0 software was used to quantify the recruitment of neutrophils in inflammatory sites of zebrafish juveniles.

[0057] like Figure 2 As shown, compared with the Ctrl group, neutrophils in the SLS group of zebrafish juveniles significantly migrated to the site of inflammation. Meanwhile, the HHG and YSYZ groups reduced the tendency of neutrophils to be recruited to the site of inflammation. Quantitative analysis results (Table 1) further indicate that different concentrations of HHG and YSYZ can significantly inhibit neutrophil migration to the site of inflammation (P<0.001), and the inhibitory effect of the YSYZ group is significantly better than that of the HHG group.

[0058] Table 1. Effects of HHG and YSYZ on neutrophil migration in an SLS-induced zebrafish inflammation model.

[0059] Group The number of neutrophils that migrate to sites of inflammation Ctrl group 1.50±1.05 SLS Group <![CDATA[22.83±2.99 ### ]]> 50 μg / mL HHG group <![CDATA[14.50±2.26 *** <!-- 5 -->]]> 100 μg / mL HHG group <![CDATA[12.17±1.17 *** ]]> 200 μg / mL HHG group <![CDATA[10.83±1.47 *** ]]> 50 μg / mL YSYZ group <![CDATA[11.83±1.17 *** ]]> 100 μg / mL YSYZ group <![CDATA[10.50±1.05 *** ]]> 200 μg / mL YSYZ group <![CDATA[8.00±1.67 *** ]]>

[0060] Note: Data are expressed as mean ± standard deviation (n=6). One-way ANOVA was used for comparisons between multiple groups, and Dunnett's multiple comparisons were used for pairwise comparisons. Compared with the Ctrl group, ### P<0.001; compared with the SLS group, *** P<0.001.

[0061] 2. Effects of HHG and YSYZ on the expression of inflammation-related genes in an SLS-induced zebrafish inflammation model

[0062] The experimental grouping and sample processing procedures were as described above. After incubation at 28℃ for 24 h, total RNA was extracted from zebrafish juveniles in each group using RNA extraction reagent. The total RNA was then reverse transcribed into cDNA using a reverse transcription kit, followed by qPCR experiments. β-actin was used as an internal reference gene to detect the expression of inflammation-related genes TNF-α, IL-1β, IL-6, and IL-8 in zebrafish juveniles.

[0063] As shown in Table 2, compared with the Ctrl group, the expression of inflammation-related genes TNF-α, IL-1β, IL-6, and IL-8 in the SLS group was significantly upregulated (P<0.001), while the expression of inflammation-related genes in zebrafish juveniles was downregulated to varying degrees in the 200 μg / mL HHG group and the 200 μg / mL YSYZ group (P<0.05, P<0.01, P<0.001). The inhibitory effect of the 200 μg / mL YSYZ group was significantly better than that of the 200 μg / mL HHG group. The results indicate that the ethyl acetate extract of this traditional Chinese medicine compound (YSYZ) can significantly alleviate the inflammatory response.

[0064] Table 2 Effects of HHG and YSYZ on the expression of inflammation-related genes in an SLS-induced zebrafish inflammation model

[0065] Group TNF-α relative expression level IL-1β relative expression level IL-6 relative expression level IL-8 relative expression level Ctrl group 1.000±0.218 1.000±0.109 1.000±0.161 1.000±0.319 SLS Group <![CDATA[11.031±1.270 ### ]]> <![CDATA[8.453±0.823 ### ]]> <![CDATA[10.571±1.499 ### ]]> <![CDATA[12.192±1.517 ### ]]> 200 μg / mL HHG group <![CDATA[7.522±1.118 * ]]> <![CDATA[6.729±0.685 * ]]> <![CDATA[7.851±0.769 ** ]]> <![CDATA[8.890±0.761 ** ]]> 200 μg / mL YSYZ group <![CDATA[5.606±0.721 *** ]]> <![CDATA[5.259±0.752 ** ]]> <![CDATA[6.817±0.843 ** ]]> <![CDATA[7.877±1.238 ** ]]>

[0066] Note: Data are expressed as mean ± standard deviation (n=3). One-way ANOVA was used for comparisons among multiple groups, and Dunnett's multiple comparisons were used for pairwise comparisons. Compared with the Ctrl group, ### P<0.001; compared with the SLS group, * P<0.05, ** P<0.01, *** P<0.001.

[0067] Example 3

[0068] This embodiment evaluates the effects of HHG and YSYZ on angiogenesis in zebrafish juveniles based on Tg (fli1:EGFP) transgenic zebrafish.

[0069] 1. Effects of HHG and YSYZ on intersegmental angiogenesis in juvenile zebrafish

[0070] Preparation of test solution: Weigh appropriate amounts of sodium dodecyl sulfate (SLS), vascular endothelial growth factor receptor tyrosine kinase inhibitor II (VRI), 70% ethanol extract (HHG), and ethyl acetate extract (YSYZ). Dissolve in an appropriate amount of DMSO, then dilute with zebrafish embryo culture medium and bring to a suitable volume to obtain the test solution. Before use, take an appropriate amount of the stock solution and dilute with culture medium to the required concentration.

[0071] Zebrafish embryos in good condition and developed to 24 h were selected. The eggshells were removed with 1 mg / mL streptomycin solution, and the embryos were placed in 24-well plates with 10 juveniles per well as one group. The Ctrl group was given zebrafish embryo culture medium containing 0.1% DMSO, the SLS group was given 50 μg / mL SLS, the VRI group was given 50 μg / mL SLS and 500 ng / mL VRI, the HHG group was given 50 μg / mL SLS and 50, 100, and 200 μg / mL HHG, respectively, and the YSYZ group was given 50 μg / mL SLS and 50, 100, and 200 μg / mL YSYZ, respectively. After incubating at 28℃ for 24 h, zebrafish juveniles were placed on biconcave slides and kept in a lateral position. The growth of intersegmental vessels (ISVs) in each group of zebrafish juveniles was observed using an upright fluorescence microscope. The number of intact and missing intersegmental vessels in each group was counted, and the ISV index was calculated. ISV index = number of intact intersegmental vessels × 1 + number of missing intersegmental vessels × 0.5.

[0072] like Figure 3 As shown, intersegmental vessels remained intact in the Ctrl group of zebrafish juveniles, while intersegmental angiogenesis in the SLS group was not significantly affected. Intersegmental angiogenesis in the VRI group was significantly inhibited, as were intersegmental angiogenesis in the HHG and YSYZ groups at different concentrations. The intersegmental vessel index of each group of zebrafish juveniles was calculated by statistically analyzing the number of intact and missing intersegmental vessels (see Table 3). The results indicate that, compared with the Ctrl group, intersegmental angiogenesis in zebrafish juveniles at different concentrations and the YSYZ group significantly inhibited intersegmental angiogenesis (P<0.001). The inhibitory effect of the YSYZ group was significantly better than that of the HHG group.

[0073] Table 3. Effects of different concentrations of HHG and YSYZ on angiogenesis in zebrafish juveniles.

[0074] Group Intersegmental vascular index Ctrl group 26.75±1.214 SLS Group 25.17±1.782 VRI Group <![CDATA[0.83±0.408 *** ]]> 50 μg / mL HHG group <![CDATA[18.83±0.817 *** ]]> 100 μg / mL HHG group <![CDATA[17.33±0.753 *** ]]> 200 μg / mL HHG group <![CDATA[15.50±1.265 *** ]]> 50 μg / mL YSYZ group <![CDATA[14.42±0.736 *** ]]> 100 μg / mL YSYZ group <![CDATA[13.17±1.366 *** ]]> 200 μg / mL YSYZ group <![CDATA[11.17±1.252 *** ]]>

[0075] Note: Data are expressed as mean ± standard deviation (n=6). One-way ANOVA was used for comparisons between multiple groups, and Dunnett's multiple comparisons were used for pairwise comparisons. Compared with the Ctrl group, *** P<0.001.

[0076] 2. Effects of HHG and YSYZ on the expression of genes related to angiogenesis in zebrafish juveniles

[0077] The zebrafish embryo processing and test sample processing procedures were as described above. After incubation at 28℃ for 24 h, total RNA was extracted from each group of zebrafish larvae using RNA extraction reagent. The total RNA was then reverse transcribed into cDNA using a reverse transcription kit, followed by qPCR experiments. β-actin was used as an internal reference gene to detect the expression of angiogenesis-related genes vegfa, flt1, kdr, and kdrl in zebrafish larvae.

[0078] As shown in Table 4, compared with the Ctrl group, there was no significant difference in the expression of angiogenesis-related genes in zebrafish larvae in the SLS group, while the expression of related genes in the VRI positive control group was significantly downregulated (P<0.001). Simultaneously, the expression of angiogenesis-related genes in zebrafish larvae in the 200 μg / mL HHG group and the 200 μg / mL YSYZ group also showed varying degrees of decrease (P<0.05, P<0.01). The inhibitory effect of the 200 μg / mL YSYZ group was significantly better than that of the 200 μg / mL HHG group. These results indicate that the ethyl acetate extract of this traditional Chinese medicine compound (YSYZ) can significantly improve angiogenesis induced by inflammatory stimulation.

[0079] Table 4. Effects of HHG and YSYZ on the expression of genes related to angiogenesis in zebrafish juveniles.

[0080] Group relative expression level of vegfa relative expression level of flt1 relative expression level of KDR kdrl relative expression level Ctrl group 1.000±0.109 1.000±0.094 1.000±0.113 1.000±0.126 SLS Group 0.828±0.131 0.816±0.103 0.811±0.083 0.793±0.076 VRI Group <![CDATA[0.038±0.019 *** ]]> <![CDATA[0.068±0.026 *** ]]> <![CDATA[0.135±0.059 *** ]]> <![CDATA[0.081±0.024 *** ]]> 200 μg / mL HHG group 0.783±0.115 <![CDATA[0.711±0.062 * ]]> <![CDATA[0.743±0.125 * ]]> <![CDATA[0.747±0.086 * ]]> 200 μg / mL YSYZ group <![CDATA[0.616±0.108 ** ]]> <![CDATA[0.582±0.116 ** ]]> <![CDATA[0.723±0.109 * ]]> <![CDATA[0.624±0.115 ** ]]>

[0081] Note: Data are expressed as mean ± standard deviation (n=3). One-way ANOVA was used for comparisons among multiple groups, and Dunnett's multiple comparisons were used for pairwise comparisons. Compared with the Ctrl group, * P<0.05, ** P<0.01, *** P<0.001.

[0082] Example 4

[0083] This embodiment uses a zebrafish behavioral analysis system to evaluate the effects of HHG and YSYZ on the movement distance of zebrafish juveniles after SLS stimulation in Example 1.

[0084] 1. Preparation of test solution

[0085] Weigh appropriate amounts of sodium dodecyl sulfate (SLS), 70% ethanol extract (HHG), and ethyl acetate extract (YSYZ) for the experiment. Dissolve them in an appropriate amount of DMSO, then dilute with zebrafish embryo culture medium and bring the volume to a suitable level to obtain the test sample stock solution. Before use, take an appropriate amount of the stock solution and dilute it with culture medium to the required concentration.

[0086] 2. Experimental grouping and sample treatment

[0087] Zebrafish larvae in good condition, 3 days post-fertilization (3 dpf), were selected and placed in 24-well plates, with 10 larvae per well as one group. The Ctrl group was treated with zebrafish embryo culture medium containing 0.1% DMSO; the SLS group was treated with 50 μg / mL SLS; the HHG group was treated with 50 μg / mL SLS and 50, 100, and 200 μg / mL HHG, respectively; and the YSYZ group was treated with 50 μg / mL SLS and 50, 100, and 200 μg / mL YSYZ, respectively. All were incubated at 28℃ until 4 dpf, pending further experiments.

[0088] 3. Behavioral analysis

[0089] Zebrafish juveniles incubated to 4 dpf were transferred to 48-well plates, one juvenile per well, and placed in a zebrafish behavior analyzer. The experimental temperature was maintained at 28℃. The experiment was conducted using ZebraLab with a duration of 10 min. The movement trajectory and distance of the zebrafish juveniles were recorded using the behavior analyzer.

[0090] The movement trajectories of each group of zebrafish fry are as follows: Figure 4 As shown in the figure, green movement paths represent movement paths with speeds of 2–6 mm / s, while red movement paths represent movement paths with speeds above 6 mm / s. Compared to the Ctrl group, the movement trajectory of the juvenile zebrafish in the SLS group was rapid and the movement distance was significantly increased (P<0.001), indicating that SLS stimulation intensified the movement of the zebrafish juveniles. Compared to the SLS group, the movement trajectory of the juvenile zebrafish in the HHG and YSYZ groups tended to be gentler and the movement distance decreased to varying degrees (P<0.01, P<0.001). Among them, the soothing effect of the YSYZ group was significantly better than that of the HHG group.

[0091] Example 5

[0092] This embodiment uses a multi-factor orthogonal experimental method to select the optimal preparation process for the ethyl acetate extract gel of traditional Chinese medicine compound.

[0093] 1. Prescription composition and preparation method

[0094] A traditional Chinese medicine compound ethyl acetate extract gel, composed of the following components (by weight percentage):

[0095] Carbomer 940 0.9%

[0096] Triethanolamine 1.0%

[0097] Low molecular weight sodium hyaluronate 0.12%

[0098] Glycerol 12.0%

[0099] Azone 2.0%

[0100] Tween 80 2.0%

[0101] Ethyl acetate extract of traditional Chinese medicine compound 4.0%

[0102] Ultrapure water balance

[0103] The preparation method of the above-mentioned gelling agent is as follows:

[0104] (1) Dissolve 0.12% low molecular weight sodium hyaluronate in 12.0% glycerol and mix well to obtain a compound moisturizer (low molecular weight sodium hyaluronate: glycerol = 1: 100). Dissolve 4.0% Chinese herbal compound ethyl acetate extract in 2.0% Tween 80 in a beaker, then add appropriate amounts of compound moisturizer and ultrapure water respectively, dissolve, then add 2.0% azone, mix well to form phase A;

[0105] (2) Add 0.9% Carbomer 940 to the remaining compound humectant, then add an appropriate amount of ultrapure water, stir to make Carbomer 940 swell and disperse (20~30 min), then stir evenly to form phase B;

[0106] (3) Add phase B to phase A, stir evenly, then add 1.0% triethanolamine, and stir continuously until gel-like to obtain gel-type cosmetic.

[0107] (4) Dispense the gel into boxes of 20 g each.

[0108] 2. Gel scoring criteria

[0109] Appearance, spreadability, viscosity, centrifugal stability, and heat and cold resistance are each scored out of 10 points, for a total of 50 points. See Table 5 for specific scoring criteria.

[0110] Table 5. Gel Scoring Criteria

[0111]

[0112] 3. Optimize the best preparation process

[0113] A three-factor, three-level orthogonal experiment was designed with carbomer 940 (factor A), glycerol (factor B), and triethanolamine (factor C) as the factors under investigation (Table 6). The preparation process was studied using the orthogonal experiment with the comprehensive score as the evaluation index (Table 7), and variance analysis was performed using SPSS 19.0 statistical software (Table 8).

[0114] Table 7 shows that the order of influence of each factor on the experimental results is C>A>B, and the optimal preparation process is A2B3C1, i.e., the amount of carbomer 940 is 0.9%, the amount of glycerol is 12.0%, and the amount of triethanolamine is 1.0%. Table 8 shows that each factor has a significant impact on the experimental results. Therefore, this condition was determined as the optimal preparation process for the gel, and gels containing 4% of the 70% ethanol extract of the traditional Chinese medicine compound, gels containing 4% of the ethyl acetate extract of the traditional Chinese medicine compound, and blank gels without 70% ethanol extract or ethyl acetate extract were prepared.

[0115] Table 6. Factor Level Table for Orthogonal Experiments

[0116] level Factor A (%) Factor B (%) Factor C (%) 1 0.6 8.0 1.0 2 0.9 10.0 2.0 3 1.2 12.0 3.0

[0117] Table 7 Orthogonal experimental design schemes and results

[0118] Test number Factor A (%) Factor B (%) Factor C (%) Overall score (points) 1 1 1 1 38.2 2 1 2 2 37.8 3 1 3 3 36.4 4 2 1 2 39.6 5 2 2 3 38.9 6 2 3 1 44.1 7 3 1 3 35.7 8 3 2 1 40.3 9 3 3 2 39.5 <![CDATA[K1]]> 112.4 113.5 122.6 / <![CDATA[K2]]> 122.6 117.0 116.9 / <![CDATA[K3]]> 115.5 120.0 111.0 / <![CDATA[K1']]> 37.47 37.83 40.87 / <![CDATA[K2']]> 40.87 39.00 38.97 / <![CDATA[K3']]> 38.50 40.00 37.00 / R 3.40 2.17 3.87 /

[0119] Table 8 Results of Orthogonal Experiment Variance Analysis

[0120] Source of variance Sum of Squares of Deviations Degrees of freedom variance P Factor A 18.229 2 9.114 0.017 Factor B 7.056 2 3.528 0.042 Factor C 22.429 2 11.214 0.014

[0121] Example 6

[0122] This embodiment evaluates the effects of topical HHG gel and YSYZ gel on inflammatory infiltration and angiogenesis in skin lesions based on an imiquimod (IMQ)-induced mouse psoriasis-like dermatitis model.

[0123] 1. Effects of topical HHG gel and YSYZ gel on imiquimod (IMQ)-induced psoriatic-like skin lesions in mice.

[0124] Imiquimod (IMQ)-induced psoriasis-like dermatitis in mice exhibits inflammatory skin phenotypes such as erythema, scaling, and inflammatory thickening. Therefore, this invention uses the IMQ-induced mouse psoriasis-like dermatitis model to evaluate the anti-inflammatory and anti-angiogenic effects of topical HHG gel and YSYZ gel.

[0125] Experimental grouping and treatment: Forty 8-week-old female BALB / c mice were randomly divided into 5 groups of 8 mice each. ①Ctrl group: normal control group; ②IMQ group: IMQ-induced mouse psoriasis-like dermatitis model group; ③KB group: IMQ-induced model + blank gel matrix test sample group; ④4% HHG gel group: IMQ-induced model + 4% HHG gel test sample group; ⑤4% YSYZ gel group: IMQ-induced model + 4% YSYZ gel test sample group. Except for the Ctrl group, all other groups of mice had 62.5 mg of 5% imiquimod cream applied daily to the hairless (3 days before formal modeling) dorsal skin (2 cm × 3 cm) for 6 consecutive days to establish the mouse psoriasis-like dermatitis model. Except for the IMQ group, each test sample group had 62.5 mg of gel applied 4 hours after IMQ application. Modeling and sample processing began on day 0 and continued until day 5, with daily photography and skin lesion scoring. On day 6, mice in each group were photographed and scored, and then skin tissue was collected from the backs of the mice. One portion was preserved in tissue fixative for HE staining, while the other portion was preserved at -80℃ for ELISA detection.

[0126] like Figure 5 As shown in Figure A, compared with the Ctrl group, mice in the IMQ and KB groups showed severe skin lesions on their backs; compared with the IMQ and KB groups, application of 4% HHG gel and 4% YSYZ gel effectively reduced skin inflammatory phenotypes such as erythema and scaling. HE staining results of skin lesions ( Figure 5 B) indicates that application of 4% HHG gel and 4% YSYZ gel significantly improved epidermal thickening and reduced inflammatory cell infiltration. Among them, the 4% YSYZ gel showed better efficacy than the 4% HHG gel.

[0127] 2. Effects of topical HHG gel and YSYZ gel on the release of inflammatory factors and the expression of angiogenesis-related factors in psoriatic lesions of mice.

[0128] Skin tissue samples stored at -80℃ were collected, subcutaneous fat was removed, and the samples were weighed, minced, and added to pre-cooled PBS at a ratio of tissue weight to PBS volume of 1:9. The homogenate was then centrifuged at 4℃ and 4000 r / min for 15 min, and the supernatant was collected. The protein concentration of each group of samples was determined using the BCA method. According to the ELISA kit instructions, the contents of tumor necrosis factor-α (TNF-α), interleukin-17A (IL-17A), vascular endothelial growth factor A (VEGF-A), and matrix metalloproteinase-9 (MMP-9) in the skin lesions were determined.

[0129] As shown in Table 9, compared with the Ctrl group, the levels of inflammatory factors TNF-α and IL-17A and angiogenesis-related factors VEGF-A and MMP-9 in the skin lesions of mice in the IMQ and KB groups were significantly increased (P<0.001). Compared with the IMQ and KB groups, the application of 4% HHG gel and 4% YSYZ gel reduced the levels of inflammatory factors and angiogenesis-related factors in the skin lesions of mice to varying degrees (P<0.05, P<0.01, P<0.001). Among them, the inhibitory effect of 4% YSYZ gel was significantly better than that of 4% HHG gel.

[0130] Table 9. Effects of topical HHG gel and YSYZ gel on the levels of inflammatory factors and angiogenesis-related factors in psoriatic lesions of mice.

[0131] Group TNF-α (pg / mg prot) IL-17A (pg / mg prot) VEGF-A (pg / mg prot) MMP-9 (ng / mg prot) Ctrl group 56.59±7.93 1.64±0.19 21.07±3.85 7.83±1.83 IMQ Group <![CDATA[101.80±11.92 △△△ ]]> <![CDATA[3.06±0.13 △△△ ]]> <![CDATA[121.20±6.28 △△△ ]]> <![CDATA[21.08±1.70 △△△ ]]> KB Group <![CDATA[108.60±29.35 △△△ ]]> <![CDATA[3.36±0.54 △△△ ]]> <![CDATA[125.10±10.22 △△△ ]]> <![CDATA[22.45±1.59 △△△ ]]> 4% HHG group 84.83±5.60 <![CDATA[2.77±0.27 * ]]> <![CDATA[86.57±4.91 ###*** ]]> <![CDATA[17.77±1.46 ##*** ]]> 4% YSYZ group <![CDATA[67.67±11.72 ##*** ]]> <![CDATA[2.39±0.15 ##*** ]]> <![CDATA[76.40±8.44 ###*** ]]> <![CDATA[16.86±1.16 ###*** ]]>

[0132] Note: Data are expressed as mean ± standard deviation (n=6). One-way ANOVA was used for comparisons between multiple groups, and Sidak's multiple comparisons were used for pairwise comparisons. Compared with the Ctrl group, △△△ P<0.001; compared with the IMQ group, ## P<0.01, ### P<0.001; compared with the KB group, * P<0.05, *** P<0.001.

[0133] This invention addresses the problems of existing anti-inflammatory and soothing skin products, such as significant side effects from chemical components, weak efficacy due to the impurities of crude herbal extracts, and poor skin feel. It successfully develops a compound ethyl acetate extract of traditional Chinese medicine with anti-inflammatory and soothing effects, along with its gel formulation. Firstly, guided by the principles of traditional Chinese medicine formulation, this invention proposes a formula with the following composition: "Sophora japonica flower as the principal ingredient, cooling the blood and directly dispelling dampness; lotus leaf as the assistant ingredient, promoting diuresis and relieving dampness; licorice root as the adjuvant ingredient, harmonizing the middle and detoxifying." This breaks away from the conventional "combination of clearing and tonifying" approach of multiple herbs, achieving a superior synergistic anti-inflammatory and soothing effect with a more concise formula, demonstrating the advanced wisdom of traditional Chinese medicine: "simple formulas with profound effects." Secondly, this invention utilizes a separation technique of "70% ethanol extraction-ethyl acetate enrichment" to obtain an extract enriched with flavonoids and alkaloids as active components. To evaluate its anti-inflammatory and soothing effects, this invention utilizes a zebrafish model animal system with a three-pronged evaluation model of "anti-inflammatory, anti-angiogenesis, and soothing behavior." This further confirms that the ethyl acetate extract (YSYZ) is superior to the unenriched 70% ethanol extract (HHG) in inhibiting neutrophil migration, downregulating the expression of key inflammatory factors such as TNF-α, inhibiting abnormal angiogenesis, and improving chemically induced dyspnea. Furthermore, this invention uses a mouse psoriasis-like dermatitis model to demonstrate that the YSYZ gel is more effective than the HHG gel in improving skin inflammation. This invention prepares the ethyl acetate extract (YSYZ) with anti-inflammatory and soothing effects into a gel that combines transdermal absorption (containing azone) with moisturizing and repairing functions (containing low-molecular-weight sodium hyaluronate and glycerol). This not only solves the drawbacks of traditional ointments such as greasiness and clogged pores, but also meets the needs of modern consumers, especially those with sensitive skin, for high-quality skincare products with its refreshing feel and excellent stability.

[0134] In summary, this invention covers the formulation of traditional Chinese medicine, extraction process, efficacy evaluation and product dosage form. The prepared compound ethyl acetate extract of traditional Chinese medicine and its gel have significant advantages in anti-inflammatory and soothing effects, safety and quality controllability, and have extremely high industrial transformation value and broad application prospects.

Claims

1. An anti-inflammatory and soothing compound ethyl acetate extract of traditional Chinese medicine, characterized in that, It is made from sophora japonica flowers, lotus leaves and licorice.

2. The ethyl acetate extract of the traditional Chinese medicine compound according to claim 1, characterized in that, It is prepared from 7-20 parts by weight of Sophora japonica flowers, 7-20 parts by weight of lotus leaves and 2-6 parts by weight of licorice.

3. A method for preparing the ethyl acetate extract of the traditional Chinese medicine compound according to claim 1 or claim 2, characterized in that, First, extract with ethanol, then perform liquid-liquid extraction with ethyl acetate.

4. The preparation method according to claim 3, characterized in that, The process includes the following steps: taking Sophora japonica flowers, lotus leaves, and licorice, adding ethanol, heating and refluxing to extract, and obtaining a filtrate; concentrating the obtained filtrate under reduced pressure, and then freeze-drying to obtain a dry extract; suspending the dry extract in water until homogeneous, adding ethyl acetate for extraction, and obtaining an extract; concentrating the obtained extract under reduced pressure, and then freeze-drying to obtain an extract.

5. The preparation method according to claim 3 or claim 4, characterized in that, The ethanol is 60-80% ethanol.

6. The use of the ethyl acetate extract of the traditional Chinese medicine compound as described in claim 1 or claim 2 in the preparation of topical preparations with anti-inflammatory and soothing effects.

7. A gel with anti-inflammatory and soothing effects, characterized in that, It includes the ethyl acetate extract of the traditional Chinese medicine compound as described in claim 1 or claim 2 and the matrix excipients.

8. The gelling agent according to claim 7, characterized in that, It is made from the following raw materials in weight percentage: 1-5% compound ethyl acetate extract of traditional Chinese medicine, 0.6-1% carbomer 940, 0.9-1.2% triethanolamine, 0.1-0.14% low molecular weight sodium hyaluronate, 10-14% glycerol, 1-3% azone and 2-4% Tween 80, with the balance being water.

9. A method for preparing the gelling agent according to claim 8, characterized in that, Includes the following steps: (1) Dissolve low molecular weight sodium hyaluronate in glycerol and mix well to obtain a compound moisturizer; dissolve the ethyl acetate extract of the traditional Chinese medicine compound in Tween 80, then add the compound moisturizer and water, dissolve and then add azone, mix well to form phase A; (2) Add Carbomer 940 to the remaining compound humectant, then add water and stir to make Carbomer 940 swell and disperse, then stir evenly to form phase B; (3) Add phase B to phase A, stir evenly, then add triethanolamine and stir until gel-like to obtain the gelling agent.

10. The use of the gel of claim 7 or claim 8 in improving skin inflammation.

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