External skin anti-inflammatory and anti-aging plant composition as well as preparation method and application thereof

Abstract

The invention relates to the field of daily chemicals, and discloses an external skin anti-inflammatory and anti-aging plant composition as well as a preparation method and application thereof. The plant composition comprises alcohol extracts of the following raw materials in parts by weight: 40-60 parts of schizonepeta spike, 20-40 parts of Chinese lobelia, 20-40 parts of ligusticum wallichii, 10-30 parts of mint and 10-30 parts of bighead atractylodes rhizome. The schizonepeta spike, the Chinese lobelia, the ligusticum wallichii, the mint and the bighead atractylodes rhizome are compounded for the first time, and the obtained mixed alcohol extract has remarkable anti-inflammatory and anti-aging effects after being externally applied to skin and can pass a biological safety test, so that the mixed alcohol extract can be used as a skin care ingredient, and is particularly used for preventing and relieving skin inflammation and resisting aging.

Description

Technical Field

[0001] The invention relates to the field of daily chemicals, and in particular to an external skin anti-inflammatory and anti-aging plant composition and a preparation method and application thereof. Background Art

[0002] The skin not only protects the body from external damage and maintains the homeostasis of body functions, but also serves as a bridge between the body and the external environment. It is in direct contact with the external environment and is more susceptible to the influence of the external environment, leading to aging. The aging of the skin, like the aging of other organs, is characterized by degenerative changes in structure and function, manifested as wrinkles, dry skin, reduced barrier integrity and thinning of the epidermis. In addition, external environmental factors such as seasonal changes, air pollution, air humidity, and ultraviolet intensity can also cause the skin to become sensitive. When these problems arise, people will perform skin care on their skin. However, due to excessive and inappropriate skin care, irritating chemicals in daily cosmetics, hair dyes and other products can also cause skin sensitivity, resulting in weakened skin resistance and enhanced skin immune response.

[0003] Inflammation refers to a series of protective responses that occur when the body's tissues are damaged by a variety of factors, such as infection or non-infection, through direct or indirect immune mechanisms. Clinical manifestations include redness, swelling, heat, pain, and itching. During the aging process of the human body, external factors cause an imbalance in the body's immune system, leading to overexpression of proinflammatory factors such as TNF-α and IL-6. Long-term stimulation of inflammatory factors can lead to a low-concentration chronic inflammatory aging state in the body, thereby increasing aging diseases.

[0004] As the largest organ and outermost protective barrier of the human body, the skin is the most direct manifestation of aging. At present, there are also related studies focusing on the relationship between inflammation and skin aging. Based on this, Franceschi first proposed the concept of "inflammatory aging (inflammaging)" in 2000. Inflammation is not only an important part of the immune system, but also has become a pathway associated with aging and aging-related diseases. It is the key to the onset of many aging-related diseases. Therefore, solving the problem of inflammation is one of the important means to delay aging.

[0005] At present, existing research at home and abroad mainly focuses on the relationship between inflammation and aging diseases, but there are relatively few studies on inflammation related to skin aging. It is worth noting that inhibiting inflammation is an effective target. It is of great significance to cut into the skin anti-aging strategy from the anti-inflammatory perspective and provide new ideas for skin rejuvenation. Therefore, there is a strong demand for plant compositions with clear effects, safety and no adverse reactions for inflammatory aging. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides an external skin anti-inflammatory and anti-aging plant composition and its preparation method and application. The present invention is the first to compound Nepeta spicata, Lobelia, Chuanxiong, Mentha and Atractylodes macrocephala, and the obtained mixed alcohol extract has significant anti-inflammatory and anti-aging effects after external application to the skin, and is safe and low in irritation, and can be used as a skin care ingredient, especially for preventing and relieving skin inflammation and anti-aging.

[0007] The specific technical scheme of the present invention is: In a first aspect, the present invention provides an external skin anti-inflammatory and anti-aging plant composition, comprising alcohol extracts of the following raw materials in parts by weight: 40-60 parts of Schizonepeta spicate, 20-40 parts of Lobelia, 20-40 parts of Ligusticum chuanxiong, 10-30 parts of mint and 10-30 parts of Atractylodes macrocephala.

[0008] In the early stage, the present invention attempted to compound a large number of different types of anti-inflammatory plant extracts. It was found that not all anti-inflammatory plant extracts can produce synergistic effects after compounding and have significant anti-inflammatory and anti-aging effects on the skin. The compatibility between different plant extracts in the system after compounding is extremely critical. If the compatibility is not good, there may even be antagonism between different plant extracts in the system, that is, the phenomenon of "1+1<1" occurs. On the other hand, during the research process, although the present invention also found that the synergistic effects of several compounded plant compositions are also very significant, the safety of most combinations in the test is relatively large in biological irritation, so they are not suitable as raw materials for skin care compositions.

[0009] Finally, the present invention screened the above formula. The present invention firstly prepared an anti-inflammatory and anti-aging plant composition by compounding the above five plant extracts and then extracting them with alcohol. Through network pharmacology research and experimental tests, we found that the compatibility between the above five specific plant extracts is excellent. After compounding them in a reasonable proportion, each plant extract can produce unexpected and significant synergistic effects in anti-inflammatory and anti-aging through more biological processes, pathways, and targets in addition to exerting their own known effects, and the combination can pass the biosafety test. In summary, the present invention shows through network pharmacology research that the above five components can work together to exert anti-inflammatory effects through more biological processes, pathways, and targets. In order to prove that the above five plant extracts are extremely compatible in skin anti-inflammatory and anti-aging, the present invention conducted a series of verification experiments: trying to replace any one of the above five plant extracts with other plant extracts with similar functions, and it was found that the compounding effect was significantly inferior to this specific combination of the present invention. Therefore, the five plant extracts in the formula of the present invention are indispensable and irreplaceable.

[0010] In addition, the present invention finds that the ratio between the above five components is also crucial to their synergistic effect. That is, not all combinations of the above components in any ratio can produce significant synergistic effects. Finally, the present invention finds that under the above preferred ratio, the overall effect of the obtained composition is the best.

[0011] Preferably, the topical skin anti-inflammatory and anti-aging botanical composition comprises the following raw materials in parts by weight: 45-55 parts of Nepeta spicate, 25-35 parts of Lobelia, 25-35 parts of Ligusticum chuanxiong, 15-25 parts of Menthus and 15-25 parts of Atractylodes macrocephala.

[0012] Most preferably, the topical skin anti-inflammatory and anti-aging botanical composition comprises the following raw materials in parts by weight: 50 parts of Nepeta spicate, 30 parts of Lobelia, 30 parts of Ligusticum chuanxiong, 20 parts of Mentha and 20 parts of Atractylodes macrocephala.

[0013] In a second aspect, the present invention provides a method for preparing the above-mentioned external skin anti-inflammatory and anti-aging plant composition, which specifically comprises the following steps: crushing the raw materials and mixing them, adding ethanol aqueous solution, soaking, and ultrasonic extraction; filtering and concentrating the obtained extract, and vacuum freeze-drying to obtain the external skin anti-inflammatory and anti-aging plant composition.

[0014] In the prior art, it is common to extract each plant separately and then mix them to make a composition. The present invention found that the composition obtained by mixing the above-mentioned 5 plants of the present invention and then extracting them at one time has better effect on skin anti-inflammatory and anti-aging. The reason may be that the main chemical components of Schizonepeta spicate are flavonoids, organic acids, terpenes, etc.; the main chemical components of Chuanxiong are; the main chemical components of Lobelia are flavonoids, alkaloids, etc.; the main chemical components of Chuanxiong are volatile oils, organic acids, alkaloids, etc.; the main chemical components of mint are volatile oils, flavonoids, terpenes, etc.; the main chemical components of Atractylodes macrocephala are volatile oils, polysaccharides, lactones, etc. In the mixed extraction process, the organic acid components promote the dissolution of components such as alkaloids by forming soluble complexes or increasing solubility in the solvent, and the polysaccharide components wrap the low-solubility flavonoid components in the nanophase by forming micelles to achieve the effect of increasing solubility. Compared with extracting them separately and then mixing them, the five plant raw materials can produce similar solubility in the mixed extraction process, and the components can promote the dissolution of each other, thus making the extract more effective.

[0015] Preferably, the concentration of the ethanol aqueous solution is 50-70wt%, and the weight is 20-40 times the total amount of the raw materials.

[0016] Preferably, the soaking time is 20-40 minutes.

[0017] Preferably, the ultrasonic extraction temperature is 50-70°C, the power is 300-500W, and the time is 0.5-2h. In a third aspect, the present invention provides the use of the above topical anti-inflammatory and anti-aging plant composition as an active ingredient in the preparation of topical anti-inflammatory and anti-aging cosmetics.

[0018] Preferably, the topical anti-inflammatory and anti-aging cosmetic is a cream, lotion, paste, ointment, mask, gel, lotion or essence.

[0019] Preferably, the external anti-inflammatory and anti-aging cosmetic further comprises an excipient.

[0020] Compared with the prior art, the beneficial effects of the present invention are: (1) The present invention is the first to prepare an anti-inflammatory and anti-aging plant composition by compounding five plant extracts, namely, Schizonepeta spicata, Lobelia chinensis, Ligusticum chuanxiong, Mentha arvense and Atractylodes macrocephala, and then extracting them through alcohol. It is found that the above five specific plant extracts have excellent compatibility with each other. After compounding them in a reasonable proportion, in addition to exerting their own known effects, each plant extract can also produce unexpected and significant synergistic effects in terms of anti-inflammatory and anti-aging.

[0021] (2) The present invention mixes the five plants and then extracts them at one time, and the obtained composition has better effects on skin anti-inflammatory and anti-aging.

[0022] (3) The plant composition of the present invention is made of pure natural raw materials, is safe, has low irritation, and has no side effects. It can be used as a skin care ingredient, especially for preventing and relieving skin inflammation and anti-aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a Venn diagram of the intersection of component targets and inflammatory targets drawn using Venny2.1.

[0024] Figure 2 This is a protein interaction map of the plant composition of the present invention on inflammatory targets.

[0025] Figure 3 The results of the chicken embryo chorioallantoic membrane test in the embodiment of the present invention are shown in the figure (the left picture is when the sample is just added, and the right picture is 5 minutes after the solution is added). DETAILED DESCRIPTION

[0026] The present invention will be further described below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, several changes and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0027] Overall embodiment Firstly, the present invention provides an external skin anti-inflammatory and anti-aging plant composition, comprising alcohol extracts of the following raw materials in parts by weight: 40-60 parts of schizonepeta spicate, 20-40 parts of lobelia, 20-40 parts of chuanxiong, 10-30 parts of mint and 10-30 parts of atractylodes.

[0028] In some preferred implementation cases, the topical skin anti-inflammatory and anti-aging plant composition comprises the following raw materials in parts by weight: 45-55 parts of Nepeta spicate, 25-35 parts of Lobelia, 25-35 parts of Ligusticum chuanxiong, 15-25 parts of mint and 15-25 parts of Atractylodes macrocephala.

[0029] In some more preferred implementation cases, the topical skin anti-inflammatory and anti-aging plant composition comprises the following raw materials in parts by weight: 50 parts of Nepeta spicate, 30 parts of Lobelia, 30 parts of Ligusticum chuanxiong, 20 parts of mint and 20 parts of Atractylodes macrocephala.

[0030] In a second aspect, the present invention provides a method for preparing an external skin anti-inflammatory and anti-aging plant composition, which specifically comprises the following steps: crushing the raw materials and mixing them, adding ethanol aqueous solution, soaking, and ultrasonic extraction; filtering and concentrating the obtained extract, and vacuum freeze-drying to obtain an external skin anti-inflammatory and anti-aging plant composition.

[0031] In some preferred implementation cases, the raw material is crushed and sieved through a 10-100 mesh sieve. The concentration of the ethanol aqueous solution is 50-70wt%, and the weight is 20-40 times the total amount of the raw material. The soaking time is 20-40min.

[0032] In some preferred implementation cases, the ultrasonic extraction temperature is 50-70°C, the power is 300-500W, and the time is 0.5-2h. In a third aspect, the present invention provides the use of the above-mentioned topical anti-inflammatory and anti-aging plant composition as an active ingredient in the preparation of topical anti-inflammatory and anti-aging cosmetics.

[0033] In some specific implementation cases, the topical anti-inflammatory and anti-aging cosmetic is a cream, lotion, paste, ointment, mask, gel, lotion or essence.

[0034] In some specific implementation cases, the topical anti-inflammatory and anti-aging cosmetic further includes an excipient. Specific embodiments (I) Network pharmacology to explore the synergistic effects of plant combinations (a) Screening of anti-inflammatory targets of plant compositions and individual components The chemical component information was obtained through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, http: / / tcmspw.com / tcmsp.php), using "Jessiebow spicate" (JJS), "Bileanthera" (BBL), "Chuanxiong" (CX), "Menthus" (BH), and "Atractylodes" (BZ) as keywords, and "oral bioavailability ≥ 30%" and "drug similarity ≥ 0.18" as screening conditions. The chemical component information was input into the Swisstarget (https: / / www.swisstargetrediction.ch) database for target prediction to obtain the action targets of each component. The Genecards (https: / / www.genecards.org) database was searched with "inflammation" as the keyword, and "Score value ≥ 10" was used as the screening condition to obtain inflammation-related targets. Through the "Venny2.1" online software, the targets of the plant composition, "Schizonepeta", "Lobelia", "Chuanxiong", "Mentha" and "Atractylodes" were intersected with the inflammatory targets respectively, and the resulting intersection was the anti-inflammatory target.

[0036] (b) Construction of PPI (protein interaction network) The anti-inflammatory targets of the plant combination, "Nepeta spicata", "Lobelia", "Ligusticum chuanxiong", "Mentha" and "Atractylodes macrocephala" were imported into the STRING (https: / / string-db.org / ) database to construct the PPI network. The protein interaction diagram of the plant combination on the inflammatory target is shown in Figure 2 The obtained data files were imported into Cytoscape software for topological analysis, and target genes with values ​​greater than the median of Degree were screened as potential core target genes.

[0037] (c) GO functional annotation and KEGG enrichment analysis The anti-inflammatory targets of the plant composite, “Schizonepeta spicate”, “Lobelia chinensis”, “Ligusticum chuanxiong”, “Mentha arvense” and “Atractylodes macrocephala” were respectively imported into the Metascape (https: / / metascape.org / ) database, and the species was set as “Homo sapiens” for GO gene ontology functional annotation analysis and KEGG pathway enrichment analysis.

[0038] Through the above research, there are 36 active ingredients that meet the requirements in the plant composition, including 8 from Nepeta spicate, 17 from Lobelia, 7 from Chuanxiong, 7 from Mentha, and 4 from Atractylodes. Among them, components A1 and A2 are common components of Nepeta spicate, Lobelia, and Mentha; B is a common component of Nepeta spicate, Chuanxiong, and Mentha; C1 and C2 are common components of Lobelia and Mentha; D is a common component of Nepeta spicate and Lobelia.

[0039] After deduplication of targets in each database, there were 205 targets for Nepeta tenuifolia, 281 for Lobelia chinensis, 323 for Ligusticum chuanxiong, 198 for Mentha arvensis, and 168 for Atractylodes macrocephala. At the same time, 152 targets related to inflammation were obtained after deduplication of each database. The intersection Venn diagram of component targets and inflammatory targets was drawn using Venny2.1, and the overlapping parts were potential anti-inflammatory targets, such as Figure 1 As shown in the figure, there were 27 targets in total for the intersection between plant compositions and inflammation, including 15 in Schizonepeta spicata, 18 in Lobelia chinensis, 17 in Chuanxiong, 16 in Mentha arvense, and 9 in Atractylodes macrocephala.

[0040] The intersection targets of the active ingredients of the plant composition and inflammation were uploaded to the Striing database to construct the PPI interaction network data file. The data file was imported into the Cytoscape3.7.1 software and calculated using its built-in tool NetworkAnalyzer to obtain 27 nodes, 203 edges, and a median degree value of 15. Nodes with a degree value ≥ 15 were taken as core target genes, and a total of 13 core targets were screened out from the plant composition, including 8 from Nepeta spicata, 10 from Lobelia, 6 from Chuanxiong, 9 from Mentha, and 3 from Atractylodes.

[0041] GO enrichment analysis mainly includes three parts: biological process, cell composition and molecular function. The targets of plant composites in biological processes mainly involve the regulation of inflammatory response, positive regulation of cell migration, positive regulation of cytokine production, inflammatory response and response to lipopolysaccharide; in cell composition, they mainly involve vesicle lumen, plasma membrane outer side, membrane rafts, extracellular matrix, endocytic vesicles, etc.; in molecular function, they mainly involve cytokine receptor binding, phosphatase binding, protease binding, protein kinase activity, etc. The targets of Nepeta spicata in biological processes mainly involve the regulation of smooth muscle cell proliferation, the regulation of inflammatory response, the cell response to lipids, the regulation of epithelial cell migration, etc.; in cell composition, they mainly involve vesicle lumen, endocytic vesicles, cytoplasmic perinuclear region, receptor complex, etc.; in molecular function, they mainly design heme binding, protein serine / threonine kinase activity, phosphatase binding, protein domain specific binding, etc. The targets of Lobelia chinensis in biological processes mainly involve the regulation of inflammatory response, the regulation of smooth muscle cell proliferation, the positive regulation of phosphorus metabolism, the regulation of epithelial cell migration, etc. In terms of cell composition, it mainly involves the vesicle lumen, endocytic vesicles, membrane rafts, cytoplasmic perinuclear region, etc. In terms of molecular function, it mainly involves cytokine receptor binding, protein kinase activity, protease binding, heme binding, etc. The targets of Chuanxiong in biological processes mainly involve inflammatory response, the regulation of inflammatory response, the positive regulation of cytokine production, leukocyte migration, etc. In terms of cell composition, it mainly involves membrane rafts, vesicle lumen, membrane sides, cytoplasmic perinuclear region, etc. In terms of molecular function, it mainly involves phosphatase binding, DNA binding transcript binding, carboxylic acid binding, cytokine receptor binding, etc. The targets of mint in biological processes mainly involve the regulation of inflammatory response, positive regulation of phosphorus metabolism, regulation of endopeptidase activity, regulation of epithelial cell migration, etc. In terms of cell composition, it mainly involves vesicle lumen, endocytic vesicles, membrane rafts, and the outer side of the plasma membrane, etc. In terms of molecular function, it mainly involves kinase regulatory factor activity, cytokine receptor activity, protease binding, heme binding, etc. The targets of Atractylodes macrocephala in biological processes mainly involve the regulation of inflammatory response, carboxylic acid transport, cell response to non-biological stimuli, and regulation of cytokine production involved in inflammatory response, etc. In terms of cell composition, it mainly involves the granular cavity rich in ficolin-1, and in terms of molecular function, it mainly involves oxidoreductase activity.

[0042] Through KEGG signaling pathway analysis, it can be found that the targets of the plant composition mainly involve Kaposi's sarcoma-associated herpes virus infection, AGE-RAGE signaling pathway in diabetic complications, cancer pathways, PI3K-Akt signaling pathway, inflammatory bowel disease, AMPK signaling pathway, Apelin signaling pathway, etc. The targets of Nepeta spicata mainly involve cancer pathways, toxoplasmosis, HIF-1 signaling pathway, MicroRNA in cancer, and AMPK signaling pathway, etc. The targets of Lobelia mainly involve cancer pathways, toxoplasmosis, Fc εRI signaling pathway, HIF-1 signaling pathway, AMPK signaling pathway, etc. The targets of Chuanxiong mainly involve Kaposi's sarcoma-associated herpes virus infection, chemokine signaling pathway, cancer pathways, PI3K-Akt signaling pathway, etc. The targets of mint mainly involve cancer pathways, AMPK signaling pathway, neutrophil extracellular trap formation, etc. The targets of Atractylodes macrocephala mainly involve IL-17 signaling pathways, cancer pathways, pinocytosis, and MicroRNA in cancer.

[0043] In summary, network pharmacology shows that the various components in the plant composition can pass through more biological processes, pathways, and targets when used together than when used alone. Different components can synergistically act on the same target to enhance efficacy, or they can act on different targets and pathways to expand the efficacy, thereby playing a synergistic role.

[0044] (II) Effects of different extraction methods on the efficacy of plant compositions Example 1 (Each raw material is first extracted separately and then mixed) A topical skin anti-inflammatory and anti-aging plant composition, the preparation method of which is as follows: 40 parts by weight of schizonepeta spicate, 20 parts by weight of lobelia, 20 parts by weight of chuanxiong, 10 parts by weight of mint, and 10 parts by weight of atractylodes macrocephala are crushed respectively, and passed through a 40-mesh sieve respectively, and each is added with 30 times the amount of 60% ethanol solution and soaked for 0.5 hours, and then ultrasonically extracted at 60°C with a power of 400W for 1 hour to obtain extracts of schizonepeta spicate, lobelia, chuanxiong, mint, and atractylodes macrocephala. The extracts of each raw material are filtered, concentrated, and vacuum freeze-dried to obtain powders of each raw material extract. The powders of each raw material extract are evenly mixed to obtain a topical skin anti-inflammatory and anti-aging plant composition.

[0045] Embodiment 2 (raw materials are first mixed and then extracted) A topical skin anti-inflammatory and anti-aging plant composition, the preparation method of which is as follows: 40 parts by weight of schizonepeta spicate, 20 parts by weight of lobelia, 20 parts by weight of chuanxiong, 10 parts by weight of mint, and 10 parts by weight of atractylodes macrocephala are crushed, passed through a 40-mesh sieve, mixed, added with 30 times the amount of 60% ethanol solution, soaked for 0.5 hours, and extracted at 60°C with an ultrasonic power of 400W for 1 hour. The extract is filtered, concentrated, and vacuum freeze-dried to obtain the topical skin anti-inflammatory and anti-aging plant composition.

[0046] Data comparison (a) DPPH free radical scavenging assay To evaluate the antioxidant efficacy of the botanical composition, a DPPH free radical scavenging assay was performed.

[0047] Experimental method: Dissolve each sample in anhydrous ethanol to prepare a 10 mg / mL sample solution. Take 1 mL of sample solution and mix with an equal volume of 2*10 -4 mol / L DPPH ethanol solution, mix thoroughly, react at room temperature in the dark for 0.5h, and measure the absorbance at 517nm using an ultraviolet spectrophotometer. Calculate the DPPH free radical scavenging ability according to the following formula: DPPH free radical scavenging rate (%) = [(B + CA) / B] * 100 Wherein, A is the absorbance measured when 1 mL of sample solution is mixed with an equal volume of DPPH ethanol solution, B is the absorbance measured when 1 mL of anhydrous ethanol is mixed with an equal volume of DPPH ethanol solution, and C is the absorbance measured when 1 mL of sample solution is mixed with an equal volume of anhydrous ethanol.

[0048] (b) Collagenase inhibition test To evaluate the anti-aging efficacy of the botanical composition, a collagenase inhibition test was performed.

[0049] Experimental method: Use Tricine buffer solution to prepare 1.1U / mL collagenase solution and 1mmol / L FALGPA solution. Use pure water to prepare the plant composition into a 1mg / mL sample test solution, then add 50μL sample solution, 20μL collagenase solution and 60μL Tricine buffer solution to mix evenly, incubate at 37℃ for 20min, then add 50μL FALGPA solution, shake well, react at 37℃ for 30min, and use an enzyme reader to measure the absorbance at 335nm. The collagenase inhibition rate is calculated according to the following formula: Collagenase inhibition rate (%) = [1-(CD) / (AB)]*100 Wherein: A is the absorbance without adding the sample to be tested and adding the substrate (FALGPA); B is the absorbance without adding the sample to be tested and adding the substrate; C is the absorbance with adding the sample to be tested and adding the substrate; D is the absorbance with adding the sample to be tested and not adding the substrate.

[0050] (c) Anti-inflammatory efficacy evaluation - inflammatory factor detection To evaluate the anti-inflammatory activity of the plant composition, lipopolysaccharide was used to induce RAW264.7 mouse mononuclear macrophages to simulate the inflammation model, and the effect of the plant composition on the content of inflammatory factors TNF-α and IL-6 was detected by ELISA.

[0051] Experimental method: The plant composition was prepared into a solution with a concentration of 100 μg / mL using pure water. RAW264.7 cells in the logarithmic growth phase were taken and cultured at 3×10 5 / well inoculated in a 6-well plate. Set up a normal group (Control group), a model group (Model group) and a 100μg / mL sample concentration administration group. After 24 hours of culture, the culture medium was discarded. The normal group added culture medium, the model group added culture medium containing 1μg / mL lipopolysaccharide, and the drug administration group added culture medium containing 1μg / mL lipopolysaccharide and 100μg / mL two concentrations of samples. After continuing to culture for 24 hours, the supernatant was collected, and 3 replicates were set up for each group. According to the instructions of the Elisa kit, the content of inflammatory factors TNF-α and IL-6 was detected.

[0052] The experimental data are shown in Table 1: Table 1 sample DPPH free radical scavenging rate / % Collagenase inhibition rate / % TNF-α inhibition rate / % IL-6 inhibition rate / % Example 1 72.60 29.36 49.40 49.95 Example 2 83.36 31.74 58.02 51.63 Both Example 1 and Example 2 can effectively remove DPPH free radicals and inhibit the activity of collagenase. In addition, the plant composition can also effectively inhibit the expression of inflammatory factors TNF-α and IL-6. However, through detailed comparison (as shown in Table 1), Example 2 performs significantly better than Example 1 in these effects. Further analysis shows that the plant composition of the present invention adopts a mixed extraction method, which can play a synergistic role and greatly enhance the anti-inflammatory and anti-aging effects of the plant composition. This synergistic effect makes the plant composition more advantageous in improving skin health.

[0053] (II) Effects of different proportions of components on the efficacy of plant compositions Table 2 Weigh the raw materials of each component according to the proportion in Table 2, crush Nepeta spicate, Lobelia, Chuanxiong, Mentha, and Atractylodes macrocephala, pass through a 40-mesh sieve and mix, add 30 times the amount of 60% ethanol solution and soak for 0.5 h, then extract at 60°C with an ultrasonic power of 400 W for 1 h. Filter, concentrate, and vacuum freeze-dry to obtain a plant composition extract powder.

[0054] The plant compositions of each example in Table 2 were subjected to DPPH free radical scavenging test, collagenase inhibition test and inflammatory factor detection. The experimental results are shown in Table 3: Table 3 The experimental results of Table 3 show that the plant composition (embodiments 2-26) provided by the present invention can effectively remove DPPH free radicals, inhibit collagenase activity, and reduce the expression of inflammatory factors TNF-α and IL-6. And it can be found that when the formula of the plant composition is 50-70 parts of schizonepeta spicate, 30-50 parts of lobelia, 30-50 parts of chuanxiong, 20-35 parts of peppermint, and 20-35 parts of atractylodes macrocephala, each index performs well. However, there is no significant difference (P>0.05) in efficacy between embodiment 14, embodiment 20 and embodiment 26. Considering the principles of environmental protection and resource conservation, preferred embodiment 14 is used as the best plant composition formula.

[0055] (III) Effect of substitution of different active ingredients on the efficacy of plant compositions Comparative Example 1: It is basically the same as Example 14, except that Nepeta tenuifolia is used in the plant composition instead of Nepeta tenuifolia spike, and the dosage ratio and preparation process of the other ingredients remain the same.

[0056] Comparative Example 2 is basically the same as Example 14, except that Platycodon grandiflorum is used in the plant composition instead of Lobelia chinensis, and the dosage ratio and preparation process of the other ingredients remain the same.

[0057] Comparative Example 3: is basically the same as Example 14, except that turmeric is used instead of Chuanxiong in the plant composition, and the dosage ratio and preparation process of the other ingredients remain the same.

[0058] Comparative Example 4 is basically the same as Example 14, except that Perilla leaf is used instead of mint in the plant composition, and the dosage ratio and preparation process of the other ingredients remain the same.

[0059] Comparative Example 5: It is basically the same as Example 14, except that Atractylodes macrocephala is used instead of Atractylodes macrocephala in the plant composition, and the dosage ratio and preparation process of the other ingredients remain consistent.

[0060] The above comparative examples were subjected to DPPH free radical scavenging test, collagenase inhibition test and inflammatory factor detection. The experimental results are shown in Table 4: Table 4 sample DPPH free radical scavenging rate / % Collagenase inhibition rate / % TNF-α inhibition rate / % IL-6 inhibition rate / % Embodiment 14 93.19 40.58 80.33 79.07 Comparative Example 1 90.25 38.7 71.50 70.12 Comparative Example 2 87.99 37.12 65.43 46.69 Comparative Example 3 77.69 28.56 52.53 42.88 Comparative Example 4 84.58 29.64 67.79 53.38 Comparative Example 5 88.42 31.52 69.64 64.12 The experimental results of Table 4 show that Example 14 not only performs well in scavenging DPPH free radicals, inhibiting collagenase activity, and reducing the expression of inflammatory factors TNF-α and IL-6, but also its effect is significantly better than Comparative Examples 1-5. Further, we can find that when the 5 kinds of raw materials (Schizonepeta spicate, Lobelia, Chuanxiong, Mentha, Atractylodes macrocephala) of the plant composition in Example 14 are replaced with other raw materials with similar efficacy activities, the efficacy of the plant composition is significantly reduced. This phenomenon shows that the plant composition selected by the present invention achieves a significant synergistic effect through the interaction of these specific ingredients, and this synergistic effect greatly enhances the anti-inflammatory and anti-aging effects of the plant composition. If any component in the combination is replaced with other substances with similar functions, this synergistic effect will be destroyed.

[0061] (III) Cosmetic eye irritation test - Chicken embryo allantoic membrane test In order to evaluate the safety of the plant composition, the chicken embryo chorioallantoic membrane test was used to explore its irritation. The specific method refers to the group standard "T / SHRH 011-2018 Cosmetic Eye Irritation Test Chicken Embryo Chorioallantoic Membrane Test".

[0062] Test substance: the plant composition obtained in Example 14 at a concentration of 10 mg / mL.

[0063] Experimental method: Each sample requires 6 chicken embryos to be tested in parallel. Purchase 0-day-old chicken embryos, place them in an incubator at 37°C and 60% relative humidity for 10 days, then check and discard the unfertilized and inactive chicken embryos. Select chicken embryos with well-developed blood vessels, use tweezers to peel off the eggshell in the air chamber area, add 0.9% sodium chloride solution to fully moisten it, absorb the sodium chloride solution, and gently peel off the eggshell membrane to expose the chorioallantoic membrane. Use a pipette to add 0.3mL of the sample to be tested with a concentration of 10mg / mL to the chorioallantoic membrane, and immediately observe the reaction of the chorioallantoic membrane for 5 minutes. The experimental results are as follows: Figure 3 And as shown in Table 5.

[0064] Table 5 sample Irritation rating Stimulus evaluation Embodiment 14 0 No stimulation Result analysis: Figure 3 As shown in Table 5, the chicken embryo chorioallantoic membrane test shows that the plant composition is non-irritating, which indicates that the plant composition provided by the present invention is safe and has no adverse reaction to human skin.

[0065] (V) Human patch test In order to evaluate the safety of the plant composition, a patch test was conducted to explore its irritation by referring to the repetitive open smear test method in the 2015 "Technical Specifications for Safety of Cosmetics".

[0066] Test substance: the plant composition obtained in Example 14 at a concentration of 10 mg / mL.

[0067] Subjects: 15 females and 15 males, a total of 30 people. Age: 20 to 50 years old. Health status: The subjects had healthy skin and no history of skin allergies.

[0068] Experimental method: Pipette 20μL of the test substance and evenly drop it onto the filter paper attached to the patch tester. After placing the patch tester, apply it to the curved side of the volunteer's forearm. Use the palm of your hand to gently press it to evenly apply it to the skin. The patch tester will remain on the skin for 24 hours. Remove the patch tester for 30 minutes, and observe the skin reaction after the indentation disappears. Observe once 24 hours and 48 hours after the patch test. The evaluation criteria are as follows: normal people are negative (-); ± (0 level), suspicious, itchy or mild redness and swelling; + (1 level), weakly positive, simple erythema, itchy; ++ (2 level), moderately positive, redness and swelling, papules; +++ (3 level), strongly positive, significant redness and swelling, papules, small blisters; ++++ (4 level), extremely strong positive, significant redness and swelling, blisters, necrosis. The results are analyzed according to the reaction classification as shown in Table 6.

[0069] Table 6 sample Number of subjects Test time / h Level 0 (Example) Level 1 (Example) Level 2 (Example) Level 3 (Example) Level 4 (Example) Embodiment 14 30 24 30 0 0 0 0 Result analysis: From the experimental results in the above table, the skin of 30 subjects was negative, which shows that the plant compositions provided by the present invention are safe and have no adverse reactions to human skin.

[0070] The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0071] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. A topical skin anti-inflammatory and anti-aging plant composition, characterized in that The invention comprises alcohol extracts of the following raw materials in parts by weight: 40-60 parts of schizonepeta spicate, 20-40 parts of lobelia, 20-40 parts of chuanxiong, 10-30 parts of mint and 10-30 parts of atractylodes macrocephala.

2. The topical skin anti-inflammatory and anti-aging plant composition according to claim 1, characterized in that The invention comprises alcohol extracts of the following raw materials in parts by weight: 45-55 parts of schizonepeta spicate, 25-35 parts of lobelia, 25-35 parts of chuanxiong, 15-25 parts of mint and 15-25 parts of atractylodes macrocephala.

3. The topical skin anti-inflammatory and anti-aging plant composition according to claim 2, characterized in that The invention comprises alcohol extracts of the following raw materials in parts by weight: 50 parts of schizonepeta spicate, 30 parts of lobelia, 30 parts of chuanxiong, 20 parts of mint and 20 parts of atractylodes macrocephala.

4. A method for preparing the external skin anti-inflammatory and anti-aging plant composition according to any one of claims 1 to 3, characterized in that include: The raw materials are crushed and mixed, and ethanol aqueous solution is added, immersed, and ultrasonically extracted; the obtained extract is filtered, concentrated, and dried to obtain an external skin anti-inflammatory and anti-aging plant composition.

5. The preparation method according to claim 4, characterized in that: The concentration of the ethanol aqueous solution is 50-70wt%, and the weight is 20-40 times the total amount of the raw materials.

6. The preparation method according to claim 4, characterized in that: The soaking time is 20-40 minutes.

7. The preparation method according to claim 4, characterized in that: The temperature of the ultrasonic extraction is 50-70°C, the power is 300-500W, and the time is 0.5-2h.

8. Use of the topical anti-inflammatory and anti-aging plant composition for skin according to any one of claims 1 to 3 or the topical anti-inflammatory and anti-aging plant composition obtained by the preparation method according to any one of claims 4 to 7 as an active ingredient in the preparation of topical anti-inflammatory and anti-aging cosmetics.

9. The use according to claim 8, characterized in that: The external anti-inflammatory and anti-aging cosmetics are creams, lotions, pastes, ointments, facial masks, gels, lotions or essences.

10. The use according to claim 8 or 9, characterized in that: The external anti-inflammatory and anti-aging cosmetic further comprises an excipient.

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