Botanical composition for reducing skin damage

Abstract

The present invention relates to a plant composition for reducing skin damage, and belongs to the technical field of anti-inflammatory compositions. In the preparation process of the plant composition of the present invention, on the one hand, the alcohol precipitation and aging step removes a large number of macromolecular substances such as macromolecular polysaccharides, and on the other hand, the polyamide resin is used for adsorption in the resin adsorption step to remove flavonoids, and the obtained plant composition is stable in nature, not easy to be stratified or discolored, and is easy to store and apply to skin care products; (2) the polyamide resin is used for adsorption in the resin adsorption step to remove anthraquinone, so the obtained plant composition will not cause irritation to skin that has already suffered from inflammation, and avoids damaging the skin; (3) the D101 macroporous resin is used in the resin adsorption step to remove inorganic salts, small molecular polysaccharides and remaining impurities, and the influence of small molecular polysaccharides on the stability of the plant composition is removed. The obtained plant composition enhances the effect of inhibiting inflammation and is suitable for external skin care.

Description

Technical Field

[0001] The invention belongs to the technical field of compositions for inhibiting inflammation, and in particular, relates to a plant composition for reducing skin damage. Background Art

[0002] The skin is in direct contact with the external environment and has the functions of protection, excretion, regulating body temperature and sensing external stimuli. It is the largest organ in the human body. In daily life, the skin is constantly exposed to adverse factors such as mechanical damage, ultraviolet radiation, chemical stimulation, etc. When adverse factors break the balance of the skin, the skin will produce inflammation, causing barrier damage, skin damage and aging. Summary of the invention

[0003] The object of the present invention is to provide a plant composition for reducing skin damage, which can effectively inhibit inflammation, has stable extracted components and does not irritate the skin.

[0004] The purpose of the present invention can be achieved by the following technical solutions:

[0005] A plant composition for reducing skin damage, which is prepared from the following raw materials in parts by weight: 1 to 3 parts of dendrobium, 4 to 6 parts of swertia japonica, and 1 to 3 parts of sour orange;

[0006] The dendrobium includes plants of the genus Dendrobium in the family Orchidaceae, such as Dendrobium nobile, Dendrobium slender-stemmed, and Dendrobium striata, which contain dendrobium alkaloids;

[0007] The Swertia includes Swertia genus of Gentianaceae, such as Swertia serrata containing iridoid glycosides, Swertia japonica, Swertia india, Swertia serrata from eastern Sichuan, and Swertia serrata from green leaves;

[0008] The sour orange is fresh sour orange fruit (Citrus×aurantium (Lour.) Engl.) or Citrus aurantium L.;

[0009] The plant composition is prepared by a method comprising the following steps:

[0010] Extraction: extract the raw material with ethanol solution, filter and take the filtrate as the liquid to be treated;

[0011] Alcohol precipitation and aging: using ethanol to precipitate and age the liquid to be treated to obtain a first treated liquid;

[0012] Resin adsorption: The first treatment liquid is adsorbed with polyamide resin, and the eluate is obtained after elution with water, and then the eluate is adsorbed with D101 macroporous resin. After adsorption, the D101 macroporous resin is eluted with water and ethanol solution in sequence to obtain the second treatment liquid;

[0013] Removing microorganisms: The second treatment liquid is treated to remove microorganisms to obtain a plant composition containing total alkaloids of Dendrobium officinale, total iridoid glycosides of Swertia japonica and total alkaloids of Citrus aurantium.

[0014] As a preferred embodiment of the present invention, the volume concentration of the ethanol solution in the extraction step is 60-80%.

[0015] As a preferred embodiment of the present invention, the mass of the ethanol solution in the extraction step is 10 to 30 times that of the raw material.

[0016] As a preferred embodiment of the present invention, the mass of the ethanol in the alcohol precipitation and aging step is 1 to 3 times that of the liquid to be treated.

[0017] As a preferred embodiment of the present invention, the alcohol precipitation aging is specifically to add ethanol to the liquid to be treated, and age it at 4° C. for 12 to 48 hours to obtain the first treated liquid.

[0018] As a preferred embodiment of the present invention, the mass of the polyamide resin in the resin adsorption step is not less than 10 times that of the raw material.

[0019] As a preferred embodiment of the present invention, the mass of the D101 macroporous resin in the resin adsorption step is not less than 20 times that of the raw material.

[0020] As a preferred embodiment of the present invention, the resin adsorption step of eluting the D101 macroporous resin with water and ethanol solution in sequence is specifically to first elute the D101 macroporous resin with water until it is colorless, and then elute it with 60-70% by volume ethanol solution to obtain the second treatment solution.

[0021] As a preferred embodiment of the present invention, the second treatment liquid obtained by resin adsorption is subjected to the following operation: the second treatment liquid is concentrated to 20wt% of the total raw medicinal materials, 40-70wt% of butanediol and pure water are added to prepare an extract with a total raw medicinal material ratio of 100%, and the extract is stirred evenly.

[0022] As a preferred embodiment of the present invention, the microorganism removal treatment in the microorganism removal step includes filtration with a 0.45 μm membrane and / or sterilization.

[0023] Beneficial effects of the present invention:

[0024] In the preparation process of the plant composition of the present invention: (1) on the one hand, the alcohol precipitation and aging step removes a large amount of macromolecular substances such as macromolecular polysaccharides, and the polyamide resin is used for adsorption in the resin adsorption step to remove a part of the macromolecular polysaccharides, and basically no polysaccharide macromolecules will settle; on the other hand, the polyamide resin is used for adsorption in the resin adsorption step to remove flavonoids. After removing the macromolecular polysaccharides and flavonoids, the obtained plant composition is stable in nature, not easy to stratify or change color, and is easy to store and apply to skin care products; (2) the polyamide resin is used for adsorption in the resin adsorption step to remove anthraquinone, so the obtained plant composition will not cause irritation to the skin that has already suffered from inflammation, thereby avoiding skin damage; (3) the D101 macroporous resin is used for adsorption in the resin adsorption step to remove inorganic salts, small molecular polysaccharides and residual impurities, thereby removing the influence of small molecular polysaccharides on the stability of the plant composition. The dendrobium, swertia japonica and sour orange raw materials are prepared through the above steps to obtain a plant composition containing three components: total alkaloids of dendrobium, total cyclopentane glycosides of swertia japonica and total alkaloids of sour orange. The three components in the plant composition synergistically enhance the effect of inhibiting inflammation, and are suitable for external skin care. DETAILED DESCRIPTION

[0025] The present invention is further described in detail below in conjunction with specific implementation methods.

[0026] Example 1

[0027] A plant composition for reducing skin damage, the preparation method is as follows:

[0028] ① Extraction: extract the raw materials of Dendrobium officinale, Swertia japonica and Citrus australis in a mass ratio of 4:10:4 with an ethanol solution of 20 times the mass of the raw materials (the volume concentration of ethanol is 75%) at 75°C under stirring for 2 hours, release the liquid, filter, and concentrate to 1 times the amount of the raw medicine to obtain the liquid to be treated;

[0029] ② Alcohol precipitation aging: add 3 times the amount of crude drug in ethanol solution (ethanol volume concentration is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24h to obtain the first treated liquid;

[0030] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0031] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0032] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0033] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0034] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0035] Example 2

[0036] A plant composition for reducing skin damage, the preparation method is as follows:

[0037] ① Extraction: extract the dendrobium, swertia japonica and sour orange in a mass ratio of 2:6:2 with an ethanol solution 30 times the mass of the raw materials (the volume concentration of ethanol is 80%) at 60°C with stirring for 2 hours, release the liquid, filter, and concentrate to 1 times the amount of the raw medicine to obtain the liquid to be treated;

[0038] ② Alcohol precipitation aging: Add ethanol solution (volume concentration of ethanol is 95%) twice the amount of raw drug to the liquid to be treated, stir while adding, and age at 4°C for 48h to obtain the first treated liquid.

[0039] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0040] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0041] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0042] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 40wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0043] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0044] Example 3

[0045] A plant composition for reducing skin damage, the preparation method is as follows:

[0046] ① Extraction: extract the dendrobium, swertia japonica and sour orange in a mass ratio of 3:4:3 with an ethanol solution 10 times the mass of the raw materials (the volume concentration of ethanol is 75%) at 70°C with stirring for 2 hours, release the liquid, filter, and concentrate to 1 times the amount of the raw medicine to obtain the liquid to be treated;

[0047] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 12 hours to obtain the first treated liquid.

[0048] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0049] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0050] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 70% (volume fraction) ethanol solution to obtain the second treatment solution;

[0051] ⑥ Concentrate the second treated liquid to 20wt% of the total raw medicinal materials, add 70wt% of butanediol to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stir well, filter through a 0.45μm membrane, sterilize, and fill to obtain a plant composition for reducing skin damage.

[0052] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0053] Comparative Example 1 A plant composition for reducing skin damage

[0054] Compared with Example 1, the raw material is replaced by Dendrobium officinale, and the rest is the same as Example 1. The preparation method is as follows:

[0055] ① Extraction: extract the raw material Dendrobium with ethanol solution of 20 times the mass of the raw material (the volume concentration of ethanol is 75%) at 75°C for 2 hours with stirring, release the liquid, filter, and concentrate to 1 times the amount of the raw drug to obtain the liquid to be treated;

[0056] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0057] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0058] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0059] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0060] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0061] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0062] Comparative Example 2 A plant composition for reducing skin damage

[0063] Compared with Example 1, the raw material is replaced by Swertia japonica, and the rest is the same as Example 1. The preparation method is as follows:

[0064] ① Extraction: Use ethanol solution (volume concentration of ethanol is 75%) with 20 times the mass of the raw material to extract the raw material Swertia japonica at 75℃ for 2h, release the liquid, filter and concentrate to 1 times the amount of the raw drug to obtain the liquid to be treated;

[0065] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0066] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0067] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0068] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0069] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0070] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0071] Comparative Example 3 A plant composition for reducing skin damage

[0072] Compared with Example 1, the raw material is replaced by lime, and the rest is the same as Example 1. The preparation method is as follows:

[0073] ① Extraction: Use ethanol solution of 20 times the mass of the raw material (the volume concentration of ethanol is 75%) to extract the raw material lime at 75℃ for 2h, release the liquid, filter, and concentrate to 1 times the amount of the raw drug to obtain the liquid to be treated;

[0074] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0075] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0076] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0077] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0078] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0079] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0080] Comparative Example 4: A plant composition for reducing skin damage

[0081] Compared with Example 1, the raw materials are replaced by Dendrobium and Swertia japonica in a mass ratio of 2:5, and the rest are the same as Example 1. The preparation method is as follows:

[0082] ① Extraction: extract the Dendrobium and Swertia japonica in a mass ratio of 2:5 with an ethanol solution 20 times the mass of the raw materials (the volume concentration of ethanol is 75%) at 75°C for 2 hours with stirring, release the feed liquid, filter it, and concentrate it to 1 times the amount of the raw medicine to obtain the liquid to be treated;

[0083] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0084] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0085] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0086] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0087] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0088] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0089] Comparative Example 5: A plant composition for reducing skin damage

[0090] Compared with Example 1, the raw materials are replaced by Swertia japonica and lime in a mass ratio of 5:2, and the rest are the same as Example 1. The preparation method is as follows:

[0091] ① Extraction: Swertia japonica and sour orange in a mass ratio of 5:2 are extracted with an ethanol solution of 20 times the mass of the raw materials (the volume concentration of ethanol is 75%) at 75°C for 2 hours by stirring, the feed liquid is discharged, filtered, and concentrated to 1 times the amount of the raw drug to obtain the liquid to be treated;

[0092] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0093] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0094] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0095] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then load D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution;

[0096] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0097] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0098] Comparative Example 6 A plant composition for reducing skin damage

[0099] Compared with Example 1, the raw materials are replaced by dendrobium and orange in a mass ratio of 1:1, and the rest are the same as Example 1. The preparation method is as follows:

[0100] ① Extraction: extract the dendrobium and lime in a mass ratio of 1:1 with an ethanol solution of 20 times the mass of the raw materials (the volume concentration of ethanol is 75%) at 75°C for 2 hours with stirring, release the liquid, filter, and concentrate to 1 times the amount of the raw medicine to obtain the liquid to be treated;

[0101] ② Alcohol precipitation aging: Add 3 times the amount of raw drug in ethanol solution (the volume concentration of ethanol is 95%) to the liquid to be treated, stir while adding, and age at 4°C for 24 hours to obtain the first treated liquid.

[0102] ③ After spreading soil on the plate and frame filter, use it to filter the first treatment liquid;

[0103] ④ Concentrate the filtered first treatment liquid to 1 times the amount of crude drug, then load the polyamide resin (10 times the total amount of crude drug) and adsorb for 12 hours;

[0104] ⑤ Elute the resin with 2 column volumes of water to obtain the eluent, concentrate the eluent to 1 times the total amount of crude drug, and then add D101 macroporous resin (20 times the total amount of crude drug to load the column). The adsorbed D101 macroporous resin is first eluted with water until it is colorless, and then eluted with 2 column volumes of 60% (volume fraction) ethanol solution to obtain the second treatment solution.

[0105] ⑥ The second treated liquid is concentrated to 20wt% of the total raw medicinal materials, and 55wt% of butanediol is added to prepare an extract with a total raw medicinal material ratio of 100% (the insufficient part is supplemented with pure water), stirred evenly, filtered through a 0.45μm membrane, sterilized, and filled to obtain a plant composition for reducing skin damage.

[0106] The plant composition was detected to contain total alkaloids from Dendrobium officinale, total iridoid glycosides from Swertia japonica and total alkaloids from Citrus aurantium.

[0107] The plant compositions prepared in Examples 1 to 3 and Comparative Examples 1 to 6 were tested as follows:

[0108] Keratinocytes (provided by Guangdong Boxi Biotechnology Co., Ltd.) were stimulated with capsaicin (CAP), and the anti-inflammatory efficacy of the test samples was evaluated by detecting the changes in the content of capsaicin receptor (TRPV1). Keratinocytes were irradiated with UVB, and the anti-inflammatory efficacy of the test samples was evaluated by detecting the changes in the contents of inflammatory factors (IL-1α, TNF-α) and inflammatory mediators (PGE2).

[0109] Experimental Example 1 TRPV1 inhibition experiment

[0110] Cell inoculation: After the cells are revived, when the plating rate reaches about 60%, inoculate the cells into a 96-well plate and place them in a CO 2 Incubator (37°C, 5% CO 2 ) and incubate overnight.

[0111] Solution preparation: Prepare the working solution of the test substance according to the test plan (see Table 1).

[0112] Drug administration: According to the test grouping, set up a blank control group, a negative control group, a positive control group and a sample group. When the cell plating rate in the 24-well plate reaches 40% to 60%, group drug administration and stimulation are carried out, and each group has 3 replicate wells. Add 1 mL of culture medium to each well of the blank control group, add 1 mL of culture medium containing CAP to each well of the negative control group, add 1 mL of culture medium containing CAP and trans-4-tert-butylcyclohexanol to each well of the positive control group, and add 1 mL of culture medium containing CAP and the corresponding concentration of the sample to be tested to each well of the sample group. After drug administration, place the 24-well plate in a CO 2 Incubator (37°C, 5% CO 2 ) for 24 h.

[0113] TRPV1 detection: fix the cells with 4% paraformaldehyde for 30 minutes, then perform immunofluorescence detection of capsaicin receptor TRPV1, observe and take photos under a microscope, collect and analyze the images.

[0114] Inhibition rate calculation:

[0115] Results Statistical analysis: GraphPad Prism was used for plotting, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis among the groups, and the results are shown in Table 2. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0116] Table 1 Test plan

[0117]

[0118] Table 2 Results of inflammatory mediator (TRPV1 protein) content detection

[0119]

[0120] Note: Integrated optical density (IOD) / cell number, its value reflects the content of TRPV1. When the t-test method is used for statistical analysis, compared with the BC group, the significance is indicated by #, P-value < 0.05 is indicated by #, and P-value < 0.01 is indicated by ##; compared with the NC group, the significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0121] From the above results we can see that:

[0122] Compared with the blank control BC group, the TRPV1 protein content in the negative control NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0123] Compared with the negative control NC group, the TRPV1 protein content in the positive control PC group was significantly decreased, indicating that the positive control test was effective.

[0124] Compared with the negative control NC group, the TRPV1 protein content of the samples of Examples 1 to 3 decreased significantly.

[0125] Comparing Examples 1 to 3 with Comparative Examples 1 to 3, the inhibition rates of Examples 1 to 3 are significantly better than those of Comparative Examples 1 to 3, indicating that the plant composition is better than a single component in antagonizing TRPV1 protein, and the single use of sour orange in Comparative Example 3 has no antagonistic effect on TRPV1 protein. The inhibitory effect of Comparative Example 4 is significantly reduced compared with Examples 1 to 3, which shows that the antagonistic effect of Dendrobium officinale, Swertia japonica and sour orange on TRPV1 protein in the plant composition is better than the simple addition of Dendrobium officinale and Swertia japonica.

[0126] Test Examples 2-4 IL-1α, PGE2, TNF-α inhibition experiments

[0127] Cell inoculation: After the cells are revived, when the plating rate reaches about 60%, inoculate the cells into a 6-well plate and incubate at 4 °C for 24 h.2 Incubator (37°C, 5% CO 2 ) and incubate overnight.

[0128] Solution preparation: Prepare the working solution of the test substance according to the test plan (see Table 3).

[0129] Drug administration: According to the test, the cells were divided into blank control group, negative control group, positive control group and sample group. When the cell plating rate in the 6-well plate reached 30% to 50%, the cells were divided into groups for drug administration and stimulation, with 3 replicate wells in each group. 2 mL of culture medium was added to each well of the blank control group and the negative control group, 2 mL of culture medium containing dexamethasone was added to each well of the positive control group, and 2 mL of culture medium containing the sample to be tested at the corresponding concentration was added to each well of the sample group. After drug administration, the 24-well plate was placed in a CO 2 Incubator (37°C, 5% CO 2 ) for 24 h.

[0130] UVB irradiation: After washing the cells with PBS, all groups except the blank control group were irradiated with UVB at a dose of 300 mJ / cm 2 .

[0131] Post-incubation: After washing the cells with PBS, place the 6-well plate in a CO 2 Incubator (37°C, 5% CO 2 Incubate for 24h.

[0132] Collect cells: Collect the cell culture supernatant in a centrifuge tube and store it in a -80°C refrigerator.

[0133] ELISA test: Perform the test and analysis according to the operating instructions of the ELISA test kit.

[0134] Inhibition rate calculation:

[0135] Results Statistical analysis: GraphPad Prism was used for plotting, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis among the groups, and the results are shown in Tables 4 to 6. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0136] Table 3 Test plan

[0137]

[0138]

[0139] Table 4 Inflammatory mediator (IL-1α) content test results

[0140]

[0141]

[0142] Note: When the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P-value < 0.05 was indicated by #, and P-value < 0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0143] Compared with the blank control BC group, the IL-1α content in the negative control NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0144] Compared with the negative control NC group, the IL-1α content in the positive control PC (dexamethasone) group decreased significantly, indicating that the positive control in this test was effective.

[0145] Compared with the negative control NC group, the IL-1α content of Examples 1 to 3, Comparative Example 2, Comparative Example 3 and Comparative Example 5 groups decreased significantly.

[0146] Comparing Examples 1 to 3 with Comparative Examples 1 to 3, the inhibition rates of Examples 1 to 3 are significantly better than those of Comparative Examples 1 to 3, indicating that the plant composition is better than a single component in antagonizing IL-1α, and the single use of Dendrobium in Comparative Example 1 has no antagonistic effect on IL-1α. The inhibitory effect of Comparative Example 5 is significantly lower than that of Examples 1 to 3, which shows that the antagonistic effect of Dendrobium, Swertia japonica and Citrus aurantium on IL-1α in the plant composition is better than the simple addition of Swertia japonica and Citrus aurantium.

[0147] Table 5 Inflammatory mediator (PGE2) content test results

[0148] Experimental Grouping Average concentration (pg / mL) SD P-value Inhibition rate Blank control 179.30 10.55 / / Negative control 324.20 3.66 0.000## / Positive Control 254.89 30.37 0.017* / Example 1 270.84 17.58 0.007* 16.5％ Example 2 276.15 19.26 0.013* 14.8％ Example 3 280.47 18.16 0.015* 13.5％ Comparative Example 1 291.18 16.64 0.028* 10.2％ Comparative Example 2 469.26 32.66 0.002** / Comparative Example 3 402.18 34.91 0.018* /

[0149] Note: When the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P-value < 0.05 was indicated by #, and P-value < 0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0150] Compared with the blank control BC group, the PGE2 content in the negative control NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0151] Compared with the negative control NC group, the PGE2 content in the positive control PC (dexamethasone) group decreased significantly, indicating that the positive control in this test was effective.

[0152] Compared with the negative control NC group, the PGE2 content of Examples 1 to 3 and Comparative Example 1 group decreased significantly.

[0153] Comparing Examples 1 to 3 with Comparative Examples 1 to 3, the inhibition rates of Examples 1 to 3 are significantly better than those of Comparative Examples 1 to 3, indicating that the plant composition is better than a single component in the antagonistic effect on PGE2 protein, and the single use of deerskin in Comparative Example 2 and the single use of lime in Comparative Example 3 have no antagonistic effect on PGE2 protein.

[0154] Table 6 Inflammatory mediator (TNF-α) content detection results

[0155] Experimental Grouping Average concentration (pg / mL) SD P-value Inhibition rate Blank control 5.69 0.53 / / Negative control 8.41 0.67 0.005## / Positive Control 5.99 0.81 0.016* / Example 1 4.53 0.51 0.001** 46.1％ Example 2 4.68 0.43 0.001** 44.4％ Example 3 4.64 0.39 0.001** 44.8％ Comparative Example 1 5.68 0.27 0.003** 32.5％ Comparative Example 2 9.42 1.01 0.223 / Comparative Example 3 6.52 0.19 0.009** 22.5％ Comparative Example 6 6.21 0.25 0.006** 26.2％

[0156] Note: When the t-test method was used for statistical analysis, compared with the BC group, the significance was indicated by #, P-value < 0.05 was indicated by #, and P-value < 0.01 was indicated by ##; compared with the NC group, the significance was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0157] Compared with the blank control BC group, the TNF-α content in the negative control NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0158] Compared with the negative control NC group, the TNF-α content in the positive control PC (dexamethasone) group decreased significantly, indicating that the positive control in this test was effective.

[0159] Compared with the negative control NC group, the TNF-α content in Examples 1 to 3, Comparative Example 1, Comparative Example 3 and Comparative Example 6 groups decreased significantly.

[0160] Comparing Examples 1 to 3 and Comparative Examples 1 to 3, the inhibition rates of Examples 1 to 3 are significantly better than those of Comparative Examples 1 to 3, indicating that the plant composition is better than a single component in antagonizing TNF-α, and the use of Swertia japonica alone in Comparative Example 2 has no antagonistic effect on TNF-α. The inhibitory effect of Comparative Example 6 is significantly reduced compared to Examples 1 to 3, which shows that the antagonistic effect of Dendrobium officinale, Swertia japonica and Citrus aurantium on TNF-α in the plant composition is better than the simple addition of Dendrobium officinale and Citrus aurantium.

[0161] IL-1, TNF-α, PGE2 and TRPV1 are cytokines, inflammatory mediators and receptors closely related to skin damage, and play an important role in local skin inflammation and immune response. This program can not only reduce the inflammatory response by inhibiting the above targets when skin damage has occurred, but also inhibit the itching, redness, swelling, pain, burning sensation and other skin inflammatory responses to external stimuli for normal skin.

[0162] IL-1 exists in the form of IL-1α and IL-1β. In the epidermal cells and endothelial cells of human skin, the expression of IL-1α is very high under normal conditions. After the IL-1α gene is translated, it first exists in the cell in the form of pro-IL-1α, which is the precursor protein of IL-1α. Pro-IL-1α can enter the cell nucleus to regulate the transcription of other genes, and can also move to the cell membrane surface through glycosylation to bind to the IL-1R1 receptor to release signals, causing related downstream reactions. When the human body is stimulated and stressed, the expression of IL-1α increases, upregulating the NF-kB pathway and promoting the production of many pro-inflammatory factors such as IL-6 and IL-8. Unlike IL-1α, the expression of IL-1β needs to be guided by the immune system such as macrophages, and is not constantly expressed in healthy individuals. After the IL-1β gene is translated, it also exists in its precursor form (pro-IL-1β) until the signal released by the immune system is activated by the NLRP3 inflammasome. Since IL-1α and IL-1β release strong pro-inflammatory signals and cause imbalance in the skin ecology, many skin diseases are associated with increased expression of IL-1α and IL-1β.

[0163] Endogenous TNF is a fat-soluble glycoprotein oligomer that exists in the skin. TNF plays an important role in the inflammatory response of keratinocytes. TNF-α can induce the expression of mediators such as interleukin-1 (IL-1) and intercellular adhesion molecule-1 (ICAM-1), causing the aggregation of inflammatory cells.

[0164] TRPV1 receptors, as nociceptors of the skin, can be activated by acid, high temperature and certain chemicals, causing the release of neurogenic inflammatory factors such as calcitonin gene-related peptide CGRP and SP, inducing skin keratinocytes, IL-1, IL-8, MCP-1 and other cytokines and neuronal nitric oxide synthase, NO, inducing local capillary vasodilation, lymphocyte and neutrophil infiltration. Histamine is an important signal substance in the inflammatory signaling pathway. Once the relevant inflammatory factors activate mast cells, they produce histamine, which acts on histamine receptors, dilates capillaries, and allows immune cells to quickly reach the site of inflammation.

[0165] PG (prostaglandin) is a vasodilator synthesized by cyclooxygenase 1 (COX-1) and its isozyme COX-2. PG can directly act on nociceptors to produce pain and make tissues more sensitive to other mediators that produce inflammation and pain. It is present in the damaged skin that is healing. The damaged skin regulates the production of PGE2 by upregulating COX-2 protein content through ROS / ERK, resulting in local redness, heat, swelling and pain. PGE2 can also cause inflammation and itching of the skin exposed to sunlight, and antihistamines can partially alleviate the skin itching caused by it. PGE2 itself does not cause itching, but it can reduce the itching threshold and enhance the itching caused by histamine.

[0166] In daily life, the skin is constantly exposed to the invasion of adverse factors, such as mechanical damage, ultraviolet rays, irritating chemicals, etc. When the balance is broken, inflammation causes barrier damage, skin damage, and aging. Therefore, we expect that by experimenting and screening the raw material embodiments 1 to 3 that can inhibit IL-1, TNF-α, PGE2 and TRPV1 targets, the skin damage caused by external factors can be reduced.

[0167] The above is only an implementation method of the invention, and does not limit the scope of patent protection. Those skilled in the art can make non-substantial changes or substitutions based on the invention, which still fall within the scope of patent protection.

Claims

1. A plant composition for reducing skin damage, characterized in that: The plant composition is prepared from the following raw materials in parts by weight: 1 to 3 parts of dendrobium, 4 to 6 parts of swertia japonica, and 1 to 3 parts of sour orange; the plant composition is prepared by a method comprising the following steps: Extraction: extracting the raw material with an ethanol solution, wherein the volume concentration of the ethanol solution is 60-80%, filtering and taking the filtrate as the liquid to be treated; Alcohol precipitation aging: add ethanol to the liquid to be treated, age it at 4°C for 12 to 48 hours, and obtain a first treated liquid, wherein the mass of the ethanol is 1 to 3 times that of the liquid to be treated; Resin adsorption: The first treatment liquid is adsorbed with a polyamide resin, and the eluate is obtained after elution with water, and then the eluate is adsorbed with a D101 macroporous resin. After adsorption, the D101 macroporous resin is firstly eluted with water until it is colorless, and then eluted with a 60-70% by volume ethanol solution to obtain a second treatment liquid; the second treatment liquid is concentrated to 20wt% of the total crude drug, and 40-70wt% of butanediol and pure water are added to prepare an extract with a total crude drug ratio of 100%, and the extract is stirred evenly; Removing microorganisms: The second treatment liquid is treated to remove microorganisms to obtain a plant composition containing total alkaloids of Dendrobium officinale, total iridoid glycosides of Swertia japonica and total alkaloids of Citrus aurantium.

2. The plant composition according to claim 1, characterized in that The mass of the ethanol solution in the extraction step is 10 to 30 times that of the raw material.

3. The plant composition according to claim 1, characterized in that The mass of the polyamide resin in the resin adsorption step is not less than 10 times that of the raw material.

4. The plant composition according to claim 1, characterized in that The mass of the D101 macroporous resin in the resin adsorption step is not less than 20 times that of the raw material.

5. The plant composition according to claim 1, characterized in that The microorganism removal treatment in the microorganism removal step includes filtration with a 0.45 μm membrane and / or sterilization.

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