Soothing composition, use and skin care product

Abstract

The present invention belongs to the technical field of skin care, and discloses a soothing composition, use and skin care product. The soothing composition comprises rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract. The rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract are obtained by composite extraction of rye seeds, yellow gentian root, aubergine fruit and marshmallow root. The mass ratio of the rye seeds, yellow gentian root, aubergine fruit and marshmallow root is 1~8:1~8:0.1~3:0.1~3. The present invention adopts a compound of rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract. The compound of rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract synergistically inhibits pro-inflammatory factors, synergistically enhances the skin soothing effect, and synergistically enhances the antioxidant effect of the composition.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care, and in particular to a soothing composition, use thereof and a skin care product. Background Art

[0002] Exposing the skin to the sun can easily cause skin damage, the main reason being the direct and indirect damage of ultraviolet radiation to the skin, and the main ultraviolet rays that can cause skin damage are UVA and UVB; UVA has strong penetrating power and can reach the dermis, destroying collagen and elastic fibers, causing skin sagging and wrinkles (photoaging), and will produce reactive oxygen species (ROS), triggering oxidative stress and damaging cell DNA, lipids and proteins; UVB has high energy and mainly damages the epidermis, directly destroying DNA, causing sunburn (erythema, blisters, peeling), and activating inflammatory responses, releasing pro-inflammatory factors (such as IL-6, TNF-α), causing redness, swelling and pain.

[0003] Therefore, the skin needs timely after-sun care to avoid further damage to the skin and to avoid abnormal cell function or apoptosis. Currently, many technologies have been deeply researched and applied for skin sun protection.

[0004] For example, prior art 1: Chinese patent 200910069813.5 discloses the use of Schisandra chinensis extract in the preparation of sunscreen skin care products. Its specification conducts in vitro research on the protection of Schisandra chinensis extract against ultraviolet radiation damage, thereby clearly showing that Schisandra chinensis extract can still effectively protect skin cells under ultraviolet radiation, thereby improving the cell survival rate of the skin; prior art 1 verifies through specific implementation methods that Schisandra chinensis extract can effectively protect skin cells and avoid the problem of excessive cell damage after sun exposure leading to a significant decrease in cell survival rate.

[0005] Just like prior art 2: Chinese patent application 202411269889.3 discloses the use of Trigonella ternate extract in the preparation of external preparations for skin after-sun repair. Trigonella ternate extract can scavenge free radicals, inhibit lipid peroxidation, inhibit the production of ROS, down-regulate the expression of AGEs, and inhibit the inflammatory response induced by ultraviolet rays, reduce the expression of NF-κB and HMGB1 genes upregulated by ultraviolet rays, inhibit the secretion of inflammatory factor TNF-α, and at the same time reduce the expression of melanin-promoting factors α-MSH and β-catenin genes upregulated by ultraviolet rays, inhibit the production of melanin and the transport of melanosomes, that is, significantly reduce the symptoms of skin after-sun damage by improving multiple signal pathways such as skin oxidative damage, inflammatory response, pigmentation, etc. caused by ultraviolet rays.

[0006] Both prior art 1 and prior art 2 have demonstrated that some natural plant extracts can be used for after-sun skin repair, and cosmetics used for after-sun care can produce good results; however, since the dosage of a single component needs to be limited in the application of cosmetics, in order to further improve the after-sun soothing effect of cosmetics and achieve the effect of reducing ultraviolet damage, it is necessary to conduct in-depth research in the direction of compounding multiple natural plant extracts. This is a direction that neither prior art 1 nor prior art 2 has studied. Prior art 1 and prior art 2 only verify the skin sun protection effect of a single natural plant extract. Summary of the Invention

[0007] One of the purposes of the present invention is to provide a soothing composition to solve the problem that the soothing effect of a single component after sun exposure is limited and the soothing effect of the skin after sun exposure needs to be further improved. By compounding multiple components, a synergistically improved soothing effect of the composition after sun exposure is achieved, which can effectively reduce the content of inflammatory factors and bring excellent antioxidant effects.

[0008] Another object of the present invention is to provide a use of a soothing composition, which can be used to prepare cosmetics to provide the cosmetics with a certain effect of soothing after sun exposure and reducing ultraviolet damage.

[0009] At the same time, the present invention also provides a skin care product with excellent soothing effect after sun exposure.

[0010] To achieve the above-mentioned purpose, the present invention provides a soothing composition, comprising rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract, wherein the rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract are obtained by composite extraction of rye seeds, yellow gentian root, aubergine fruit and marshmallow root, and the mass ratio of the rye seeds, yellow gentian root, aubergine fruit and marshmallow root is 1~8:1~8:0.1~3:0.1~3.

[0011] Rye seed extract is rich in ferulic acid and natural resorcinol, both potent antioxidants. Ultraviolet (UV) radiation induces the production of reactive oxygen species (ROS) in the skin, leading to oxidative stress and cellular damage. Ferulic acid scavenges hydrogen peroxide and superoxide free radicals, reducing lipid peroxidation and protecting cell membrane integrity. Resorcinol inhibits tyrosinase activity, reducing melanin production and indirectly reducing post-sun pigmentation.

[0012] In addition, the β-glucan and polyphenols in rye seed extract have anti-inflammatory effects, which can inhibit the release of inflammatory factors (such as TNF-α and IL-6) and reduce skin inflammatory responses.

[0013] The active ingredients in Gentiana scabra root extract have anti-inflammatory activity, which can inhibit the release of inflammatory mediators and the activation of inflammatory cells, thereby reducing the inflammatory response of the skin; by inhibiting the activation of inflammatory signaling pathways such as nuclear factor-κB (NF-κB), it reduces the production of inflammatory factors such as interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α), thereby relieving the redness and pain of the skin after sun exposure and promoting skin repair; at the same time, Gentiana scabra root extract helps restore the skin's elasticity and firmness by promoting the synthesis of collagen, accelerating the repair and regeneration of damaged skin after sun exposure.

[0014] Passion fruit extract is rich in powerful antioxidant ingredients such as vitamin C, flavonoids, SOD enzyme, etc., which can neutralize the excess free radicals produced in the skin after ultraviolet radiation and reduce oxidative stress damage; and the potassium contained in the passion fruit extract helps regulate the skin microenvironment and relieve post-sun sensitivity.

[0015] Some ingredients in marshmallow root extract, such as mucin and flavonoids, have anti-inflammatory properties; flavonoids can inhibit the release of inflammatory mediators, such as interleukins and prostaglandins, which are produced in large quantities after the skin is exposed to ultraviolet rays, triggering an inflammatory response; by inhibiting their release and activity, marshmallow root extract can reduce inflammatory symptoms such as redness and pain of the skin, and help the skin return to normal; mucin can form a breathable protective film on the skin surface to prevent excessive evaporation of moisture, while also absorbing moisture in the air to replenish the skin and keep the skin moist; good skin moisture status helps the metabolism and self-repair of skin cells, and promotes the recovery of skin after sun exposure.

[0016] The synergistic mechanism of the present invention may be due to the following reasons: by compounding rye seed extract, gentiana root extract, passion fruit extract and marshmallow root extract, the skin condition is synergistically regulated from the aspects of anti-inflammation, anti-oxidation, regulating skin microenvironment, and maintaining skin moisture, thereby effectively reducing the content of various inflammatory factors in the skin after sun exposure, and synergistically improving the antioxidant effect of the composition, so that the composition exhibits excellent soothing effect after sun exposure.

[0017] Preferably, the composite extraction of the rye seed extract, the gentian root extract, the passion fruit extract and the marshmallow root extract comprises the following steps:

[0018] Step 1: Add rye seeds, yellow gentian root, aubergine fruit, and marshmallow root to a 50% ethanol solution, extract at 50-60°C for 3-4 hours, and centrifuge to obtain an extract, wherein the weight of the 50% ethanol solution is 1-20 times the total weight of the rye seeds, yellow gentian root, aubergine fruit, and marshmallow root;

[0019] Step 2: The extract is transferred to a falling film concentrator, and the filtrate is vacuum concentrated at a concentration temperature of 55-65°C and a concentration pressure of -0.08-0.1 MPa to zero ethanol content to obtain an alcohol-free concentrate;

[0020] Step 3: Add butanediol and deionized water to the alcohol-free concentrate, mix and stir evenly, then add the mixture to an ultrafiltration device loaded with an ultrafiltration membrane with a molecular weight cutoff of 150 to 500 kDa, and perform ultrafiltration at an ultrafiltration pressure of 0.1 to 0.3 MPa to obtain an ultrafiltrate, wherein the amount of butanediol added is 10 to 70% of the total weight, and the amount of deionized water added is 0 to 85% of the total weight.

[0021] Furthermore, the composite extraction of rye seed extract, gentiana root extract, passion fruit extract and marshmallow root extract also includes step 4: adding a preservative to the ultrafiltrate and sterilizing it.

[0022] Preferably, the preservative is at least one of p-hydroxyacetophenone, 1,2-hexanediol, caprylhydroxamic acid, pentylene glycol, ethylhexylglycerin, caprylyl glycol, phenoxyethanol, and sodium benzoate.

[0023] The invention also discloses use of the soothing composition in preparing cosmetics.

[0024] Preferably, the dosage forms of the cosmetics include lotions, creams, emulsions, essences, liquid foundations and liquid masks.

[0025] The present invention also discloses a skin care product containing 0.1 to 25 wt% of the soothing composition.

[0026] Beneficial effects

[0027] Compared with the prior art, the present invention has at least the following advantages:

[0028] The present invention adopts a compound of rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract. Not only does the compound of rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract synergistically inhibit pro-inflammatory factors, synergistically enhance the soothing effect of skin after sun exposure, but also synergistically enhance the antioxidant effect of the composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The present invention will be further described below with reference to the accompanying drawings and embodiments;

[0030] Figure 1 This is a diagram of a chick embryo before using the soothing composition of Example 4 of the present invention;

[0031] Figure 2 This is a picture of a chick embryo after using the soothing composition of Example 4 of the present invention. DETAILED DESCRIPTION

[0032] The present invention will be further described below in conjunction with the embodiments, but this does not constitute any limitation to the present invention. Any limited modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0033] In order to explain the technical content of the present invention in detail, further description will be given below in conjunction with the embodiments.

[0034] In the following examples and comparative examples, the rye seeds were purchased from a grain store in the market; the gentian root was purchased from a Chinese medicine store, the egg fruit was purchased from a Chinese medicine store, and the marshmallow root was purchased from Bozhou Yuanshengtang Pharmaceutical Co., Ltd.

[0035] It should be noted that the technical solution of the present application can be implemented through other commercially available products and is not limited to the raw materials obtained from the above sources.

[0036] In the following examples and comparative examples, unless otherwise specified, parts are parts by weight and % is percentage by weight.

[0037] Example 1

[0038] A soothing composition is prepared by the following steps:

[0039] Step 1: Add 3 parts of rye seeds, 5 parts of yellow gentian roots, 1 part of passion fruit, and 0.1 parts of marshmallow roots to a 50% ethanol solution with a concentration of 10 times the total weight of rye seeds, yellow gentian roots, passion fruit, and marshmallow roots, extract at a temperature of 55±2°C for 3 hours, and centrifuge to obtain an extract;

[0040] Step 2: The extract is transferred to a falling film concentrator, and the filtrate is vacuum concentrated at a concentration temperature of 60° C. and a concentration pressure of -0.08 MPa to zero ethanol content to obtain an alcohol-free concentrate;

[0041] Step 3: adding butanediol and deionized water to the alcohol-free concentrate, stirring the mixture evenly, and then adding the mixture to an ultrafiltration device loaded with an ultrafiltration membrane with a molecular weight cutoff of 300 kDa, and ultrafiltration was performed at an ultrafiltration pressure of 0.2 MPa to obtain an ultrafiltrate, wherein the amount of butanediol added was 30% of the total weight, and the amount of deionized water added was 20% of the total weight.

[0042] Example 2

[0043] It is generally the same as Example 1, except that: in step 1, the portions of rye seeds, gentian roots, egg fruit, and marshmallow roots are 8 portions, 1 portion, 0.5 portion, and 3 portions, respectively.

[0044] Example 3

[0045] It is generally the same as Example 1, except that: in step 1, the portions of rye seeds, gentian roots, egg fruit, and marshmallow roots are 1.5 portions, 8 portions, 3 portions, and 0.5 portions, respectively.

[0046] Example 4

[0047] It is generally the same as Example 1, except that: in step 1, the amounts of rye seeds, gentian roots, egg fruit, and marshmallow roots are 4 parts, 4 parts, 1 part, and 1 part, respectively.

[0048] Comparative Example 1

[0049] It is generally the same as Example 4, except that step 1 is changed to: add 4.45 parts of rye seeds, 4.45 parts of yellow gentian roots, and 1.1 parts of aubergine to an ethanol solution with a concentration of 50% that is 10 times the total weight of rye seeds, yellow gentian roots, and aubergine, extract at a temperature of 55±2°C for 3 hours, and centrifuge to obtain an extract.

[0050] Comparative Example 2

[0051] The method is substantially the same as Example 4, except that step 1 is changed to: adding 4.45 parts of rye seeds, 4.45 parts of yellow gentian roots, and 1.1 parts of marshmallow roots to a 50% ethanol solution having a concentration of 10 times the total weight of rye seeds, yellow gentian roots, and marshmallow roots, extracting at a temperature of 55±2° C. for 3 h, and centrifuging to obtain an extract.

[0052] Comparative Example 3

[0053] It is generally the same as Example 4, except that step 1 is changed to: add 7.2 parts of yellow gentian root, 1.4 parts of aubergine, and 1.4 parts of marshmallow root to 50% ethanol solution with a concentration of 10 times the total weight of yellow gentian root, aubergine, and marshmallow root, extract at a temperature of 55±2°C for 3h, and centrifuge to obtain an extract.

[0054] Comparative Example 4

[0055] It is generally the same as Example 4, except that step 1 is changed to: add 7.2 parts of rye seeds, 1.4 parts of aubergine, and 1.4 parts of marshmallow roots to an ethanol solution with a concentration of 50% that is 10 times the total weight of rye seeds, aubergine, and marshmallow roots, extract at a temperature of 55±2°C for 3 hours, and centrifuge to obtain an extract.

[0056] Efficacy testing

[0057] 1. UVB-induced inflammatory factor IL-6 content in human keratinocytes

[0058] 1. Experimental Purpose and Principle

[0059] The skin is the immune organ most vulnerable to damage after UVB exposure. UVB can directly or indirectly induce the release of cytokines from skin cells, leading to skin immunosuppression. Keratinocytes are UVB-sensitive target cells that, after UVB exposure, secrete a variety of cytokines, such as IL-6, which participate in various inflammatory responses.

[0060] 2 Test indicators

[0061] IL-6 relative content judgment standard: IL-6 relative content is lower than that of negative control and has a large difference with negative control.

[0062] 3 Experimental materials and methods

[0063] 3.1 Instruments: RT-6100 enzyme-labeled analyzer; ultraviolet lamp (302 nm).

[0064] 3.2 Cells used in the experiment: keratinocytes.

[0065] 3.3 Test methods

[0066] (1) Treatment of test samples

[0067] Sample group: The sample was diluted with pure water to a 4% solution, then filtered with a 0.22 μm filter, and the filtrate was collected as the sample mother solution; the sample mother solution was then further diluted with pure water to a sample concentration of 0.5%;

[0068] Negative control: basal medium.

[0069] (2) Test operation steps

[0070] IL-6 content determination: cells were plated in a 96-well plate and cultured for 24 hours. Culture medium containing the above sample at a concentration of 0.5% was added to the sample wells, culture medium was added to the negative control wells, and culture medium was added to the normal control wells. The cells were incubated for 24 hours. The supernatant was discarded, and the above cells were rinsed three times with PBS. The normal control group was completely wrapped with tin foil and placed in a dark place. The cells in the other irradiation groups were placed under a UVB lamp with an irradiation dose of 90mJ / cm 2 After completion, the PBS solution was discarded and the culture medium was added respectively. After incubation for 4 hours, the supernatant was aspirated and the IL-6 content in the cell culture supernatant was determined using an ELISA kit to obtain the effect of the test sample on the IL-6 content of keratinocytes.

[0071] (3) Calculation formula

[0072] .

[0073] The compositions of Examples 1-4 and Comparative Examples 1-4 were tested according to the above method. The results are shown in Table 1.

[0074] Table 1 Test results of relative IL-6 content of the compositions of Examples 1-4 and Comparative Examples 1-4

[0075] Sample name IL-6 relative content (%) Example 1 88.35 Example 2 87.49 Example 3 86.78 Example 4 85.22 Comparative Example 1 94.20 Comparative Example 2 96.38 Comparative Example 3 98.75 Comparative Example 4 98.86

[0076] According to the results in Table 1, we can see that:

[0077] According to the data of Examples 1-4, the present application can effectively inhibit the release of IL-6 and reduce the inflammatory response of the IL-6 pathway by combining rye seed extract, yellow gentian root extract, passion fruit extract and marshmallow root extract. Even if the sample concentration is only diluted to 0.5%, it has a relatively obvious effect of inhibiting IL-6 release; and there will be differences depending on the amount of each component used. In terms of the effect of reducing the content of the inflammatory factor IL-6, Example 1 is more effective;

[0078] According to the data comparison between Example 4 and Comparative Examples 1-4, it can be seen that when any one of the rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract is lost, even if the total weight of the composition is the same, the effect of inhibiting IL-6 release shown by the composition will be significantly reduced, indicating that Comparative Examples 1-4 only produce the efficacy superposition between the components; especially Comparative Examples 3 and Comparative Examples 4, their IL-6 relative content is relatively high and close to the negative control, because Comparative Example 3 loses the rye seed extract and Comparative Example 4 loses the yellow gentian root extract. Since the rye seed extract and the yellow gentian root extract themselves have a more significant effect of inhibiting IL-6 release, and the aubergine fruit extract and marshmallow root extract have weaker inhibitory ability on the IL-6 pathway, Comparative Examples 3 and Comparative Examples 4 are difficult to show the effect of inhibiting IL-6 release under the simple efficacy superposition of the composition at a test concentration of 0.5%; it is explained that the present application produces a synergistic inhibitory effect on IL-6 release by compounding rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract, and none of the four extracts can be missing.

[0079] 2. UVB-induced inflammatory factor TNF-α content in human keratinocytes

[0080] 1. Experimental Purpose and Principle

[0081] The skin is the immune organ most vulnerable to damage after UVB exposure. UVB can directly or indirectly induce the release of cytokines from skin cells, leading to skin immunosuppression. Keratinocytes are UVB-sensitive target cells that, after UVB exposure, secrete a variety of cytokines, such as TNF-α, which participate in various inflammatory responses.

[0082] 2 Test indicators

[0083] Criteria for determining the relative content of TNF-α: The relative content of TNF-α is lower than that of the negative control and has a significant difference from the negative control.

[0084] 3 Experimental materials and methods

[0085] 3.1 Instruments: RT-6100 enzyme-labeled analyzer; ultraviolet lamp (302 nm).

[0086] 3.2 Cells used in the experiment: keratinocytes.

[0087] 3.3 Test methods

[0088] (1) Treatment of test samples

[0089] Sample group: The sample was diluted with pure water to a 4% solution, then filtered with a 0.22 μm filter, and the filtrate was collected as the sample mother solution; the sample mother solution was then further diluted with pure water to a sample concentration of 1%;

[0090] Negative control: basal medium.

[0091] (2) Test operation steps

[0092] TNF-α content determination: Cells were plated in 96-well plates and cultured for 24 hours. Culture medium containing the above-mentioned sample at a concentration of 1% was added to the sample wells, culture medium was added to the negative control wells, and culture medium was added to the normal control wells. The cells were incubated for 24 hours. The supernatant was discarded, and the cells were rinsed three times with PBS. The normal control group was completely wrapped in tin foil and placed in the dark. The cells in the other irradiation groups were placed under a UVB lamp at an irradiation dose of 90mJ / cm. 2 After completion, the PBS solution was discarded and the culture medium was added respectively. After incubation for 4 hours, the supernatant was aspirated and the TNF-α content in the cell culture supernatant was determined using an ELISA kit to obtain the effect of the test sample on the TNF-α content of keratinocytes.

[0093] (3) Calculation formula

[0094] .

[0095] The compositions of Examples 1-4 and Comparative Examples 1-4 were tested according to the above method. The results are shown in Table 2.

[0096] Table 2 Test results of relative TNF-α content of the compositions of Examples 1-4 and Comparative Examples 1-4

[0097] Sample name Relative content of TNF-α (%) Example 1 89.33 Example 2 86.65 Example 3 88.88 Example 4 87.47 Comparative Example 1 98.13 Comparative Example 2 94.31 Comparative Example 3 95.52 Comparative Example 4 95.90

[0098] According to the results in Table 2, we can see that:

[0099] According to the data of Examples 1-4, the present application can effectively inhibit the release of TNF-α and reduce the inflammatory response of the TNF-α pathway by combining rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract. Even if the sample concentration is only diluted to 1%, it has a relatively obvious effect of inhibiting TNF-α release.

[0100] According to the data comparison between Example 4 and Comparative Example 1, in the absence of marshmallow root, the relative content of TNF-α in Comparative Example 1 is greatly increased, and there is no significant difference from the negative control, indicating that in the technical scheme of the present application, marshmallow root extract is a very important component; at the same time, combined with the data comparison of Examples 1-4, it can be seen that in terms of the effect of inhibiting the release of TNF-α, the present invention uses marshmallow root extract as the main active ingredient, and is combined with rye seed extract, gentiana root extract, and aubergine extract to synergistically enhance the effect of the composition in inhibiting the release of TNF-α.

[0101] According to the data comparison of Example 4 and Comparative Example 2-4, in the absence of any one of rye seed extract, yellow gentian root extract, and egg fruit extract, even if the total weight of the composition is the same, and Comparative Example 2-4 increases the amount of marshmallow root compared to Example 4, the relative content of TNF-α obtained by the test in Comparative Example 2-4 is still significantly improved compared to Example 4. Combining the data comparison of Example 4 and Comparative Example 1, it can be inferred that:

[0102] In terms of inhibiting the release of TNF-α, the present application uses marshmallow root extract as the core ingredient, and rye seed extract, gentiana root extract, and aubergine extract as synergistic enhancing components to synergistically enhance the inhibitory effect of marshmallow root extract on the TNF-α pathway.

[0103] At the same time, according to the comparison of the results in Table 1 and Table 2, it can be seen that the synergistic inhibition of IL-6 release produced by the combination of rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract in this application is better than the synergistic inhibition of TNF-α release, indicating that the technical solution of this application has different synergistic effects on the inhibition of different inflammatory factors.

[0104] 3. UVB-induced inflammatory factor IL-8 content in human keratinocytes

[0105] 1. Experimental Purpose and Principle

[0106] The skin is the immune organ most vulnerable to damage after UVB exposure. UVB can directly or indirectly induce the release of cytokines from skin cells, leading to skin immunosuppression. Keratinocytes are UVB-sensitive target cells that, after UVB exposure, secrete a variety of cytokines, such as IL-8, which participate in various inflammatory responses.

[0107] 2 Test indicators

[0108] IL-8 relative content test standard: The IL-8 relative content is lower than that of the negative control and has a large difference with the negative control.

[0109] 3 Experimental materials and methods

[0110] 3.1 Instruments: RT-6100 enzyme-labeled analyzer; ultraviolet lamp (302 nm).

[0111] 3.2 Cells used in the experiment: keratinocytes.

[0112] 3.3 Test methods

[0113] (1) Treatment of test samples

[0114] Sample group: The sample was diluted with pure water to a 4% solution, then filtered with a 0.22 μm filter, and the filtrate was collected as the sample mother solution; the sample mother solution was then further diluted with pure water to a sample concentration of 1%;

[0115] Negative control: basal medium.

[0116] (2) Test operation steps

[0117] IL-8 content determination: Cells were plated in 96-well plates and cultured for 24 hours. Culture medium containing 1% sample was added to the sample wells, culture medium was added to the negative control wells, and culture medium was added to the normal control wells. The cells were incubated for 24 hours. The supernatant was discarded, and the cells were rinsed three times with PBS. The normal control group was completely wrapped with tin foil and placed in the dark. The cells in the other irradiation groups were placed under a UVB lamp with an irradiation dose of 90mJ / cm 2 After completion, the PBS solution was discarded and the culture medium was added respectively. After incubation for 4 hours, the supernatant was aspirated and the IL-8 content in the cell culture supernatant was determined using an ELISA kit to obtain the effect of the test sample on the IL-8 content of keratinocytes.

[0118] (3) Calculation formula

[0119] .

[0120] The compositions of Examples 1-4 and Comparative Examples 1-4 were tested according to the above method. The results are shown in Table 3.

[0121] Table 3 Test results of relative IL-8 content of the compositions of Examples 1-4 and Comparative Examples 1-4

[0122] Sample name IL-8 relative content P-value Example 1 87.99 ＜0.05 Example 2 86.14 ＜0.05 Example 3 89.06 ＜0.05 Example 4 87.70 ＜0.05 Comparative Example 1 94.20 ＜0.05 Comparative Example 2 96.38 ＜0.05 Comparative Example 3 98.75 ＜0.05 Comparative Example 4 98.86 ＜0.05

[0123] According to the results in Table 3, we can see that:

[0124] According to the data of Examples 1-4, the present invention uses a combination of rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract to inhibit the release of IL-8.

[0125] According to the data comparison between Example 4 and Comparative Examples 1-4, it can be seen that when any one of the rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract is lost, the effect of inhibiting IL-8 release shown by the composition will also decrease significantly, indicating that Comparative Examples 1-4 only produce a superposition of the effects between the components; it shows that the present application produces a synergistic inhibitory effect on IL-8 release through the combination of rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract, and none of the four extracts can be missing.

[0126] 4. H2O2-induced fibroblast-SOD activity

[0127] 1. Experimental Purpose and Principle

[0128] The free radical theory of aging posits that cumulative free radical damage is a major cause of skin aging. With aging, free radicals accumulate within skin tissue, attacking biofilms, proteins, and nucleic acids, causing oxidative damage. Wrinkles are an outward manifestation of aging. Fibroblasts were induced with hydrogen peroxide (H2O2). The efficacy of the test substance in increasing cellular superoxide dismutase (SOD) activity was evaluated by comparing the negative control with the test substance after administration. SOD activity was measured using a SOD activity assay kit (microassay).

[0129] 2 Test indicators

[0130] Criteria for determining fibroblast SOD activity: The SOD enzyme activity of the test sample is higher than that of the negative control and is significantly different from the negative control.

[0131] 3 Experimental materials and methods

[0132] 3.1 Instrument: RT-6100 enzyme-linked microplate analyzer.

[0133] 3.2 Cells used in the experiment: Fibroblasts.

[0134] 3.3 Test methods

[0135] (1) Treatment of test samples

[0136] Sample group: basal culture medium containing 0.5 mM H2O2 and 1% sample;

[0137] Negative control group: basal culture medium containing 0.5 mM H2O2.

[0138] (2) Test operation steps

[0139] Fibroblast SOD activity detection test: 3T3 cells were plated in a 12-well plate and cultured for 24 hours. The sample group was replaced with a basal medium containing H2O2 and the samples of Examples 1-4 and Comparative Examples 1-4. The negative control group was replaced with a basal medium containing H2O2. After 24 hours of culture, the culture medium in the wells was removed and washed once with PBS. 200 μL of RIPA lysis buffer was added to each well and the cells were placed on ice to lyse for 1 minute. 1 mL of PBS was added and rinsed repeatedly for 2 minutes. The lysate was transferred to a 1.5 mL centrifuge tube and centrifuged at 1000 rpm for 3 minutes. The supernatant was collected and the SOD activity in the cell extract supernatant was measured using a kit to determine the effect of the test sample on the SOD activity of fibroblasts.

[0140] (3) Calculation formula

[0141] ; ;

[0142] Where: A0—absorbance value of control group;

[0143] A—Absorbance value of sample group.

[0144] The compositions of Examples 1-4 and Comparative Examples 1-4 were tested according to the above method. The results are shown in Table 4.

[0145] Table 4 SOD activity test results of the compositions of Examples 1-4 and Comparative Examples 1-4

[0146] Sample name SOD activity (U / ml) Example 1 5.45 Example 2 5.66 Example 3 5.82 Example 4 5.39 Comparative Example 1 5.02 Comparative Example 2 4.75 Comparative Example 3 4.67 Comparative Example 4 4.62 Negative control 1.54

[0147] According to the results in Table 4:

[0148] It should be noted that although the H2O2-induced fibroblast-SOD activity test is commonly used to test the anti-wrinkle efficacy of cosmetics, in the present invention, this test mainly reflects the anti-free radical efficacy of the composition. The core purpose of anti-free radical is to neutralize the excessive free radicals produced under ultraviolet stimulation, avoiding their continuous oxidative damage to skin cells, thereby preventing problems such as photoaging, inflammatory reactions and pigmentation.

[0149] According to the data of Examples 1-4, the present application can effectively improve the SOD activity of fibroblasts by combining rye seed extract, gentiana root extract, passion fruit extract and marshmallow root extract, and compared with the negative control, the improvement of the SOD activity of fibroblasts is significant.

[0150] According to the data comparison of Example 4 and Comparative Examples 1-4, the present application adopts the combination of rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract and does not produce a very obvious synergistic effect on the SOD activity of fibroblasts; however, according to the data of Comparative Examples 1-4, it can be known that in the composition of the present application, rye seed extract, yellow gentian root extract and aubergine fruit extract have a greater effect on the SOD activity of fibroblasts, but the improvement of fibroblast SOD activity shown in Comparative Example 4 is still weaker than that in Example 4, so it can be inferred that: marshmallow root extract Although the extract has no obvious effect on enhancing the SOD activity of fibroblasts, when it is compounded with rye seed extract, gentiana root extract and aubergine extract, it can produce the effect of assisting in enhancing the activity of rye seed extract, gentiana root extract and aubergine extract. That is to say, the compound of rye seed extract, gentiana root extract, aubergine extract and marshmallow root extract used in this application still has a certain synergistic effect on enhancing the SOD activity of fibroblasts, but it is relatively not so obvious. Compared with Comparative Example 1, the improvement of fibroblast SOD activity in Example 4 is still more than 6%.

[0151] 5. Irritation

[0152] Test method: Chicken embryo chorioallantoic membrane test

[0153] 1. Test purpose and principle

[0154] The chick chorioallantoic membrane test is an early in vitro method for evaluating eye irritation. The chorioallantoic membrane (CAM) is a respiratory membrane that surrounds the chick embryo. This test utilizes the intact, clear, and transparent vascular system of the chorioallantoic membrane of mid-stage incubated chick embryos. A certain amount of the test substance is directly exposed to the CAM. After a period of exposure, changes in toxic effect indicators (such as bleeding, coagulation, and vascular melting) are observed. These indicators reflect changes in the morphology, structure, color, and permeability of the blood vessels and vascular network, as well as phenomena such as chorioallantoic membrane protein denaturation and the degree of damage. These indicators are then combined to generate a score used to assess the eye irritation potential of the test substance.

[0155] The purpose of this test is to test the ability of the test substance to cause toxic changes in the chicken embryo chorioallantoic membrane and to evaluate the factors and processes of the potential eye irritation of the evaluated substance.

[0156] 2. Experimental Group Treatment and Reagents

[0157] Sample group: The oil-controlling and soothing composition prepared in Example 4 was diluted into a 2% aqueous solution;

[0158] Negative control: 0.9% saline;

[0159] Positive control: 1% sodium dodecyl sulfate (SDS) solution.

[0160] 3. Experimental steps

[0161] Six chick embryos were used in each group for this test. The chorioallantoic membrane (CAM) was recorded using a camera. A polytetrafluoroethylene (PTFE) ring was placed on the CAM and photographed. The sample to be tested was added to the ring, the time of sample addition was recorded, and the air chamber was covered with moistened plastic wrap. The embryos were then transferred to a constant temperature and humidity chamber for incubation, and the extent of each toxic effect was observed.

[0162] 4. Calculation of results

[0163] The endpoint evaluation method was used to calculate the endpoint evaluation (ES) with the result rounded to two decimal places. Each chicken embryo was scored as the sum of the observed bleeding, coagulation and vascular lysis. The mathematical sums obtained for ES-6 chicken embryos were averaged.

[0164] The average ES score is calculated using the following formula:

[0165] .

[0166] 5. Result judgment criteria

[0167] ES≤4, non-irritating;

[0168] 4<ES≤12, mild irritation;

[0169] 12<ES<16, moderate irritation;

[0170] ES ≥ 16, strong irritant / corrosive.

[0171] The sample of Example 4 was tested according to the above test method. The ES was less than 4, and the result was determined to be non-irritating.

[0172] Wherein, the results of chicken embryo before and after use in Example 4 are as follows Figure 1 、 Figure 2 As shown;

[0173] This proves that the technical solution of Example 4 of the present invention is mild and non-irritating.

[0174] Application Example 1

[0175] A soothing raw material liquid, the formula is shown in Table 5, and the preparation method is as follows:

[0176] Take a small amount of phase B (accounting for 20±5% of the total mass of phase B), add phase A into it, heat to 55±5℃ and stir until the solid is completely dissolved, cool to room temperature and mix with the remaining phase B to obtain a soothing raw material solution.

[0177] Table 5: Soothing raw material liquid formula

[0178]

[0179] Application Example 2

[0180] According to the formula table in Table 6, Application Example 2 is prepared as follows:

[0181] 1. Do a good job in sanitation and cleaning, and strictly clean and disinfect equipment and utensils;

[0182] 2. Pre-fabrication phase: Dissolve phase B in advance and stir to mix evenly; Mix phase C in advance and stir to mix evenly, set aside;

[0183] 3. Add the pure water in phase A and start homogenization. While homogenizing, add the remaining ingredients in phase A in sequence. Homogenize at medium speed for 60 seconds. Stir and heat to 82±3°C. Homogenize at medium speed for 2 minutes until there are no gel particles. Keep warm for 30 minutes and then stir to dehumidify.

[0184] 4. Cool down to 50℃, add phase B and phase C in sequence, and stir evenly;

[0185] 5. Cool to 45°C, add phase D and stir well.

[0186] Table 6 Formulation of Application Example 2

[0187]

[0188] The embodiments presented herein are merely embodiments selected from a combination of all possible embodiments. The appended claims should not be limited by the embodiments describing the present invention. Some numerical ranges used in the claims include subranges therein, and variations in these ranges should also be covered by the appended claims.

Claims

1. A soothing composition, characterized in that The invention comprises rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract, wherein the rye seed extract, yellow gentian root extract, aubergine fruit extract and marshmallow root extract are obtained by composite extraction of rye seeds, yellow gentian root, aubergine fruit and marshmallow root, and the mass ratio of the rye seeds, yellow gentian root, aubergine fruit and marshmallow root is 1-8:1-8:0.1-3:0.1-3; The composite extraction of the rye seed extract, the gentian root extract, the passion fruit extract and the marshmallow root extract comprises the following steps: Step 1: adding rye seeds, yellow gentian roots, aubergine fruit, and marshmallow roots to a 50% ethanol solution, extracting at a temperature of 50-60°C for 3-4 hours, and centrifuging to obtain an extract, wherein the weight of the 50% ethanol solution is 1-20 times the total weight of the rye seeds, yellow gentian roots, aubergine fruit, and marshmallow roots; Step 2: The extract is transferred to a falling film concentrator, and the filtrate is vacuum concentrated at a concentration temperature of 55-65°C and a concentration pressure of -0.08-0.1 MPa to zero ethanol content to obtain an alcohol-free concentrate; Step 3: adding butanediol and deionized water to the alcohol-free concentrate, mixing and stirring evenly, and then adding the mixture to an ultrafiltration device loaded with an ultrafiltration membrane with a molecular weight cutoff of 150-500 kDa, and ultrafiltration is performed at an ultrafiltration pressure of 0.1-0.3 MPa to obtain an ultrafiltrate, wherein the amount of butanediol added is 10-70% of the total weight, and the amount of deionized water added is 0-85% of the total weight.

2. The soothing composition according to claim 1, characterized in that The method further comprises step 4: adding a preservative to the ultrafiltrate and sterilizing the ultrafiltrate.

3. The soothing composition according to claim 2, characterized in that The preservative is at least one of p-hydroxyacetophenone, 1,2-hexanediol, caprylhydroxamic acid, pentylene glycol, ethylhexylglycerin, caprylyl glycol, phenoxyethanol, and sodium benzoate.

4. Use of the soothing composition according to any one of claims 1 to 3 in preparing cosmetics.

5. Use of the soothing composition according to claim 4 for preparing cosmetics, characterized in that: The dosage forms of the cosmetics include water, cream and lotion.

6. Use of the soothing composition according to claim 4 for preparing cosmetics, characterized in that: The cosmetics include essence, liquid foundation and liquid mask.

7. A skin care product, characterized in that: Contains 0.1 to 25 wt% of the soothing composition according to any one of claims 1 to 3.

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