CN114712280A

CN114712280A - Gel with relieving effect, preparation method thereof and evaluation method of relieving effect

Info

Publication number: CN114712280A
Application number: CN202210360648.4A
Authority: CN
Inventors: 渠志灿; 姚丽平; 李建丽; 郝花花
Original assignee: Shanxi Na'an Health Technology Co ltd
Current assignee: Shanxi Na'an Health Technology Co ltd
Priority date: 2022-04-07
Filing date: 2022-04-07
Publication date: 2022-07-08

Abstract

The invention provides gel with a relieving effect, a preparation method thereof and an evaluation method of the relieving effect, and relates to the technical field of cosmetics. The gel with the relieving effect provided by the invention utilizes the plant components to provide the relieving effect, and is safe and non-irritant. The invention also provides an evaluation method of gel soothing efficacy, which takes iNOS expression quantity as an index.

Description

Gel with relieving effect, preparation method thereof and evaluation method of relieving effect

Technical Field

The invention belongs to the technical field of cosmetics, and particularly relates to gel with a relieving effect, a preparation method of the gel and an evaluation method of the relieving effect.

Background

In recent decades, the cosmetic purchasing population has increased rapidly, and the demand for cosmetics and personal care products developed for sensitive skin consumers has been on a markedly increasing trend. The sensitive skin refers to a high-response state of the skin under physiological or pathological conditions, mainly occurs on the face, and is clinically manifested as subjective symptoms such as burning, stabbing pain, pruritus and tightness of the skin easily appearing when the skin is stimulated by physical, chemical and mental factors, and objective signs such as erythema, scale and telangiectasia are accompanied or not accompanied. An epidemiological investigation of 1039 people in the united states revealed that 69% of women and 64% of men are generally sensitive skins. From 2015 to 2019, the proportion of facial skin care products "suitable for sensitive muscles" rose from 9% to 28.7%. Consumers believe that the sensitive muscles are caused by seasonal changes, genetically sensitive skin, environmental pollution, excessive cleanliness, use of hormone-containing products, excessive exfoliating, incomplete makeup removal and the like, and buy products suitable for sensitive muscles when skin itching, facial erythema, flushing, stinging and the like occur. But the quality of products suitable for sensitive muscles is not uniform at present.

Disclosure of Invention

In view of the above, the invention aims to provide gel with a relieving effect, a preparation method thereof and an evaluation method of the relieving effect, and the relieving effect is achieved by using brand-new plant component compounding. The invention also provides a brand-new evaluation method for the soothing effect, the detection method is simple, and the detection indexes are few.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides gel with a relieving effect, and the components of the gel comprise plant extracts, water, a humectant, a thickening agent and a preservative;

the plant extracts include peony root bark extract, purslane extract, centella asiatica extract, tea extract, and feverfew extract.

Preferably, the mass of the plant extract is 0.05-6.1% of that of the jelly;

and the mass ratio of the peony root bark extract to the purslane extract to the centella extract to the tea extract to the feverfew extract is (0.01-0.1): (0.01-3.0): (0.01-1.0): (0.01-1.0): (0.01-1.0).

Preferably, the humectant is selected from one or more of glycerin, propylene glycol, sodium hyaluronate, glyceryl polyether-26 and methyl glucose polyether-20;

the mass ratio of the glycerol to the propylene glycol to the sodium hyaluronate to the glyceryl polyether-26 to the methyl glucose polyether-20 is (0.01-5): (0.01-3): (0.01-0.1): (0.01-5): (0.01-10).

Preferably, the mass of the humectant is 0.05-23.1% of the mass of the jelly.

Preferably, the thickening agent is selected from one or two of xanthan gum and sodium polyacrylate;

the preservative is selected from one or more of caprylyl hydroximic acid, glyceryl caprylate, 1, 2-hexanediol and butanediol.

The invention also provides a preparation method of the jelly, which comprises the following steps: mixing water, humectant and thickener, and stirring to obtain a first stirred material;

mixing the first stirred material and the plant extract, and performing second stirring to obtain a second stirred material;

and mixing the second stirred material with a preservative, and carrying out third stirring to obtain the jelly.

Preferably, the stirring frequency of the first stirring is 35HZ, and the stirring time is 80 min;

the stirring frequency of the second stirring is 20HZ, and the stirring time is 10 min;

the stirring frequency of the third stirring is 15HZ, and the stirring time is 15 min.

The invention also provides an evaluation method of the gel soothing effect, which comprises the following steps: by using an inflammatory cell model and taking iNOS expression quantity as an index, when the iNOS expression quantity is obviously lower than that of a negative control after gel is applied, the gel has the effect of relieving.

Preferably, the model of inflammatory cells comprises a model of inflammation established by stimulating macrophage raw264.7 with lipopolysaccharide.

Preferably, the indicators further include inflammatory states of the body and changes in skin moisture.

Has the advantages that: the invention provides gel with a relieving effect, which provides the relieving effect by utilizing plant components, is safe and has no stimulation. Paeonol in the peony root bark extract can inhibit TNF-alpha induced fibroblasts from secreting IL-1 beta and IL-8 and MMP-9 expression; the purslane extract has excellent antibacterial and anti-inflammatory effects, and alkaloids and flavones in the extract have obvious inhibition effects on common pathogenic skin fungi in human bodies, so that capillary permeability can be reduced, and inflammation and edema can be relieved; the asiaticoside extract has the main active components of madecassoside and asiaticoside, the asiaticoside can promote the synthesis of type I collagen and type III collagen, the madecassoside can promote the synthesis of laminin-5 and promote the regeneration of skin, so the asiaticoside extract has three super-strong repairing effects: scar repair, dermis repair and sunburn repair; the invention also provides an evaluation method of gel soothing effect, which takes iNOS expression quantity as an index. NO is an important inflammation medium, the generation of NO in macrophages is regulated by inducible iNOS genes, iNOS is induced and generated in inflammatory reaction in tissues to generate a large amount of NO and promote the inflammatory reaction, such as vasodilatation congestion, edema, cytotoxicity and the like, so that the relieving effect of a sample to be tested is evaluated by using the change of the expression of the iNOS genes.

In the embodiment of the invention, the gel is subjected to effect evaluation by using the evaluation method, and when the concentration of the gel is 0.63%, 0.31% and 0.16%, the iNOS gene expression level of macrophages is remarkably reduced, and the effect is remarkably different from that of a negative control group, so that the gel has a relieving effect.

Drawings

FIG. 1 shows the moisture content values within 6 hours after the product is used;

FIG. 2 is the change of the water content within 6 hours after the product is used;

FIG. 3 shows the value of the percutaneous water loss within 6 hours after the product is used;

FIG. 4 is the change rate of the percutaneous water loss value within 6 hours after the product is used;

FIG. 5 is a graph of the inhibition of cyclooxygenase-2 by PC-celecoxib;

FIG. 6 is a relative cell motility diagram;

FIG. 7 shows the relative expression levels of iNOS genes after different treatments.

Detailed Description

In the present invention, the plant extracts are soothing effect ingredients, and the plant extracts are preferably all aqueous extracts, and are preferably purchased from conventional commercial products in the art, such as peony root bark extract purchased from Beijing Jia leaf Biotech limited under the trade name of cortex moutan extract (GerbexSuffrutinosa); the herba Portulacae extract is purchased from Bailander, Korea under the trade name Portulacacatexct; centella asiatica extract is purchased from Beijing Jia leaf Biotech limited, under the trade name asiaticoside-II; tea leaf extract and feverfew extract were purchased from seiner (korea) under the trade name FKNT 30.

The weight of the plant extract is preferably 0.05-6.1% of that of the jelly; and the mass ratio of the peony root bark extract to the purslane extract to the centella extract to the tea extract to the feverfew extract is preferably (0.01-0.1): (0.01-3.0): (0.01-1.0): (0.01-1.0): (0.01-1.0), more preferably 0.03:1:0.2:0.2: 0.1; in the examples, commercially available plant red-dispelling factor NT30 comprising tea extract and feverfew extract was selected, wherein the plant red-dispelling factor NT30 comprises water, butylene glycol, 1, 3-propanediol, lecithin, feverfew extract, tea extract and magnolol in a mass ratio of 55:10:10:5:5:10:5, and was purchased from floridae chemical co.

The humectant of the present invention is preferably selected from one or more of glycerin, propylene glycol, sodium hyaluronate, glyceryl polyether-26 and methyl glucose polyether-20; the mass of the humectant is preferably 0.05-23.1% of that of the jelly. In the present embodiment, glycerin, propylene glycol, sodium hyaluronate, glyceryl polyether-26 and methyl glucose polyether-20 are used as the humectant, but they are not considered as the full protection scope of the present invention. In the embodiment of the invention, the mass ratio of the glycerol, the propylene glycol, the sodium hyaluronate, the glycerol polyether-26 and the methyl glucose polyether-20 is preferably (0.01-5): (0.01-3): (0.01-0.1): (0.01-5): (0.01-10).

The mass of the thickening agent is preferably 0.02-1.1% of that of the jelly, the thickening agent is preferably selected from one or two of xanthan gum and sodium polyacrylate, in the embodiment, commercially available sodium carbomer (MG 40R) and xanthan gum are used as the thickening agents, and the ratio of sodium polyacrylate in MG 40R: silica: the mass ratio of water is 94.5:0.5: 5.

The mass of the preservative is preferably 0-0.4% of that of the jelly, the preservative is preferably one or more of caprylyl hydroximic acid, glyceryl caprylate, 1, 2-hexanediol and butanediol, in the embodiment, E1001 is selected as the preservative, and the mass ratio of the caprylyl hydroximic acid, the glyceryl caprylate, the 1, 2-hexanediol and the butanediol in the E1001 is 12.5:3.75:3.75:80, and the preservative is purchased from Yile Biotechnology (Guangzhou) Limited.

The stirring is preferably carried out in a stirring pot, the stirring frequency of the first stirring is preferably 35HZ, and the stirring time is preferably 80 min; the stirring frequency of the second stirring is preferably 20HZ, and the stirring time is preferably 10 min; the stirring frequency of the third stirring is preferably 15HZ, the stirring time is preferably 15min, and after the thickening agent is added, the rotating speed needs to be increased, and the frequency needs to be increased.

The invention tests the relieving effect from the perspective of gene, and increases the evaluation means of the relieving test. NO is an important inflammation medium, the generation of NO in macrophages is regulated by inducible iNOS genes, iNOS is induced and generated in inflammatory reaction in tissues to generate a large amount of NO and promote the inflammatory reaction, such as vasodilatation congestion, edema, cytotoxicity and the like, so that the relieving effect of a sample to be tested is evaluated by using the change of the expression of the iNOS genes.

The inflammation cell model preferably comprises an inflammation model established by stimulating macrophage Raw264.7 by Lipopolysaccharide (LPS), and the relieving efficacy of a sample to be detected is evaluated by detecting the change of related iNOS gene expression after the sample acts.

The index for evaluating the soothing effect of the invention preferably also comprises the inflammation state of the body and the change of skin moisture, wherein the inflammation state of the body preferably comprises the activity inhibition capability of cyclooxygenase-2 (COX-2), and the soothing effect of the product is evaluated by testing the activity of the relevant enzyme COX-2 for inflammatory mediator synthesis, and the method is low in cost and high in speed compared with the method for evaluating the clinical responsiveness of a human body to a stimulant and inhibiting the generation of inflammatory-related cytokines. The COX-2 is a bifunctional enzyme with the activities of cyclooxygenase and peroxidase, is hardly expressed or has little expression amount in a normal physiological state, can be induced and expressed only when macrophages, fibroblasts, endothelial cells, monocytes and the like generate inflammation, and can be generally used as a detection index for reflecting the inflammation state of an organism. In the presence of a cofactor, COX-2 epoxidizes a substrate such as arachidonic acid using its cyclooxygenase activity to produce an intermediate such as PGG2, and COX-2 catalyzes an intermediate such as PGG2 using its peroxidase activity to produce a final product such as PGH2, and catalyzes a cyclooxygenase-2 probe that is substantially free of fluorescence to produce a probe that is strongly fluorescent (Ex560/Em 590). The enzymatic activity of COX-2 can be detected very sensitively by fluorescence detection. When COX-2 inhibitor is added to the reaction system, the fluorescence generation is inhibited, and the fluorescence intensity is inversely proportional to the inhibition effect of the inhibitor, so that the inhibition effect of the inhibitor can be detected, and the result can reflect the anti-inflammatory function of the object to be detected.

In the present invention, the skin moisture change preferably includes skin moisture content (MMV) and skin moisture dispersion, wherein skin moisture content: the stratum corneum structure of the skin forms a natural barrier to water loss, which is lost transdermally when the skin barrier function is compromised and the stratum corneum is not sufficiently hydrated to allow the skin to become drier. The method for testing the moisture content of the skin is preferably a skin capacitance method, a tester of the method is a Corneometer capacitance tester, moisture is a substance with the largest dielectric constant on the skin, and after the moisture is contacted with the skin through a test probe, the change of the capacitance value reflects the moisture content in the stratum corneum of the skin.

The skin water dispersion loss of the invention is as follows: the transepidermal water loss rate can be used to detect the amount of transepidermal water loss (TEWL) in the stratum corneum of the skin, and when the skin barrier function is disrupted, the TEWL value increases. A common testing instrument is a Tewameter instrument, a relatively stable testing small environment is formed on the surface of skin by using a cavity measuring probe with an open design based on Fick diffusion law, and the water vapor pressure gradient of two different points formed by the water loss of the horny layer close to the epidermis is measured by two groups of temperature and humidity sensors to measure the water component evaporated by the epidermis.

When the jelly prepared by the method is evaluated by using the evaluation method, the iNOS gene expression level of macrophages is remarkably reduced when the concentrations of the jelly products are 0.63%, 0.31% and 0.16%, and the jelly has a remarkable difference with a negative control group and has a relieving effect; when the concentration of the jelly is 0.125%, the activity inhibition rate of the cyclooxygenase-2 can reach about 60%, and the jelly has the activity inhibition capacity of the cyclooxygenase-2; within 6h after the gel is used, compared with a control group, the MMV of an experimental group is higher than that of the control group in a time period, the MMV change rate of the experimental group is more than 30%, compared with the control group, the MMV change rate has extremely significant difference (P < 0.001), and the MMV change rate is in negative correlation with time within 4 h. Given that glycerin is a raw material with a good moisturizing effect, the MMV in an experimental group is higher than that of glycerin within 2 hours and is almost as high as that of glycerin within 4-6 hours, which shows that the sample has the effect of remarkably increasing the moisture content of the stratum corneum of the skin and has a good moisturizing effect within 6 hours; within 6h after the product is used, TEWL in the experimental group is lower than that in the control group in the same time period, TEWL change rates are negative values, the TEWL change rates in the 6h test group are greater than 20%, and the TEWL change rates have very significant difference (P < 0.001) compared with that in the control group. Compared with the effect of glycerol, the TEWL change rate is greater than that of the glycerol within 1-6 h, which shows that the sample can obviously improve the water retention capacity of the skin cuticle and the effect is superior to that of the glycerol. The jelly has good moisture retention effect, and the water retention capacity is superior to that of glycerin (3% glycerin aqueous solution).

The gel having soothing effect and the preparation method and the evaluation method of soothing effect thereof according to the present invention will be described in detail with reference to the following examples, which should not be construed as limiting the scope of the present invention.

Example 1

86.22 parts by mass of water, a humectant (2 parts by mass of glycerin, 1.5 parts by mass of propylene glycol, 0.025 parts by mass of sodium hyaluronate, 262 parts by mass of glyceryl polyether-262 parts by mass of and 204 parts by mass of methyl glucose polyether) and a thickener (0.025 parts by mass of xanthan gum and 0.5 parts by mass of MG 40R) are mixed and subjected to first stirring to obtain a first stirred material;

mixing the first stirred material with plant extracts (0.03 parts by mass of peony root-bark extract, 0.2 parts by mass of asiaticoside-II, 1 part by mass of purslane extract and 302 parts by mass of plant red-removing factor NT) and carrying out second stirring to obtain a second stirred material;

and mixing the second stirred material with a preservative (E10010.5 parts by mass) for third stirring to obtain the gel, namely the Lu' an ice-moistening water-storage essence gel.

Example 2

Skin moisture content and transdermal moisture loss test

Materials and methods

1. And (3) measuring an object: lu Anna ice-moistening water-storage essence gel.

2. Subject: the total of 30 people, 9 men and 21 women, the age of 18 to 65 years, meet the volunteer selection criteria of the subjects.

3. Testing parts: the palmar parts of the left and right forearms.

4. Testing time points:

short-term moisturizing effect: before product use (T0), 1h with product (T1), 2h with product (T2), 4h after product (T3) and 6h after product (T4).

5. An analytical instrument:

5.1 determination of skin moisture content (MMV) instrument: a skin moisture meter Corneometer CM 825 is used for detecting the moisture content of stratum corneum of skin at arm parts. The higher the Corneometer value, the higher the moisture content of the stratum corneum;

5.2 determination of transepidermal Water loss Rate (TEWL) instruments: the transepidermal water loss rate test probe TewameterTM300 is used for detecting the transepidermal water loss rate of the skin at the arm part. Higher TEWL values indicate poorer skin barrier function and less water-holding capacity.

Second, testing step

1. The subject goes to the laboratory according to the specified time, gently rubs and cleans the arm test part by using the amino acid cleansing foam, then washes the arm test part by using clear water, does not need to rub the skin forcefully in the whole process, and lightly dips the arm test part to be dry by using absorbent paper.

2. After cleaning, the test piece enters an efficacy laboratory, and background data collection is carried out after sitting statically for 30 minutes. Then coating a sample to be tested on the test part, coating each area for ten times by using fingers to circle at a constant speed, and then starting timing.

3. Data need to be collected for 1h, 2h, 4h and 6h after sample application, and each test needs to enter an efficacy laboratory for half an hour in advance for sitting still.

Third, test results

1. Within 6h after the product is used, compared with a control group (blank), the MMV of the experimental group is higher than that of the control group (see figure 1 and table 1), the MMV change rate of 6h is more than 30%, compared with the control group, the MMV change rate has a very significant difference (P < 0.001), and the MMV change rate is in negative correlation with time within 4h (see figure 2 and table 2). The fact that glycerin is known as a raw material with a good moisturizing effect, MMV in an experimental group is higher than that of glycerin within 2 hours and is almost the same as that of glycerin within 4-6 hours indicates that the sample has the effect of remarkably increasing the moisture content of the skin stratum corneum and has a good moisturizing effect within 6 hours;

2. the TEWL values in the experimental group were all lower than the control group (see FIGS. 3 and 3) for 6h after the product was used, and the TEWL change rates were all negative, and the 6h change rates were all greater than 20%, with a very significant difference compared to the control group (P < 0.001, see FIGS. 4 and 4). Compared with the effect of glycerol, the TEWL change rate is greater than that of the glycerol within 1-6 h, which shows that the sample can obviously improve the water retention capacity of the skin cuticle and the effect is superior to that of the glycerol.

3. In conclusion, the Lu Anna ice-moistening water-storage essence gel has a good moisture-retaining effect, and the water-retaining capacity of the Lu Anna ice-moistening water-storage essence gel is superior to that of glycerin (a 3% glycerin water solution).

TABLE 1 moisture content values of the product after 6h

TABLE 2 moisture content Change within 6h after use of the product

Table 3: the product has percutaneous water loss value within 6h after use

TABLE 4 Change rate of the percutaneous Water loss value within 6h after the product was used

Example 3

Screening assay for cyclooxygenase-2 inhibitors

First, experiment method

2. Setting the initial concentration of a measured object: 10%, the product was diluted 10 times.

3. A dilution mode: 2 times of dilution, and setting high, medium and low concentrations.

4. The instrument comprises the following steps: a low-speed centrifuge, a vortex mixing instrument, a fluorescence microplate reader, an analytical balance and a pipettor.

Second, result analysis

The standard equation of the PC standard (celecoxib) is obtained by using EILSA Calc software to perform regression curve analysis (figure 5):

y＝18.55+1.35x-0.009x² R²＝0.989

and calculating to obtain: IC (integrated circuit)₅₀＝28.55nM。

The Luanna ice-moistening water-storage essence gel has the activity inhibition capacity of the cyclooxygenase-2, when the concentration is 0.5%, the activity inhibition rate of the cyclooxygenase-2 is low, but the inhibition rate is obviously increased after the gel is diluted by 2-4 times and can reach about 60% (table 5). Probably because the product components are complex, the fluorescence value of the system is influenced under the condition of high concentration, but the interference factor on the fluorescence value is weakened after dilution. The above results can be obtained: the Luanna ice-moistening water-storage essence gel can achieve an anti-inflammatory effect by inhibiting the activity of cyclooxygenase-2.

TABLE 5 inhibition of cyclooxygenase-2 Activity

Example 4

Anti-inflammatory Gene expression iNOS Gene test

First, experimental material

1. Test system

The cell used in this test was macrophage raw264.7 (mouse macrophage cell line), purchased from cell bank of chinese academy of sciences.

2. Reagent

High-glucose DMEM medium (Gibco), fetal bovine serum (Gibco), pbs (Gibco), mtt (sigma), DMSO (national medicine), trypsin (Gibco).

3. Main equipment

CO₂Incubator (Thermo, 160i), biosafety cabinet (ESCO, LA2-6a1), inverted microscope (come card, DMi8), microplate reader (Tecan, Spark), micro-oscillator (linbel, TS-92).

4. Sample information

The Luanna ice-moistening water-storage essence gel is liquid and water-soluble and is stored at 4 ℃.

Second, Experimental methods

1. Cytotoxicity test

1) Cell inoculation: by 1 × 10⁴Cell/well inoculation Density cells were plated onto 96 well plates, incubators (37 ℃, 5% CO)₂) And incubated overnight.

2) Grouping experiments: the experiment set up zero-set, solvent control group, positive control group and sample group. In the sample set, 8 concentration gradients were set for each sample, and 3 replicate wells were set for each concentration gradient.

3) Preparing liquid: sample working solutions of different concentrations were prepared according to the test concentration setting table (table 6).

TABLE 6 test concentration setting table

4) Administration: and (3) administration is carried out when the cell plating rate in the 96-well plate reaches 40-60%. Adding 200 mu L of culture solution into each well of the solvent control group; adding 200 mu L of culture solution containing 10% DMSO into each well of the positive control group; adding 200 mu L of culture solution containing samples with corresponding concentrations into each well of the sample group; the null-adjusted group was seeded without cells, and only 200. mu.L of cells were added.

5) And (4) a culture solution. After completion of the administration, the 96-well plate was placed in an incubator (37 ℃ C., 5% CO)₂) Culturing in medium.

6) And (3) detection: after 24h of cell incubation culture, the supernatant was discarded, MTT working solution (0.5mg/mL) was added, incubation was performed at 37 ℃ for 4h in the dark, after the incubation was completed, the supernatant was discarded, 150. mu.L of DMSO was added to each well, and OD was read at 490 nm.

Calculating the relative activity of the cells: according to a formula to calculate

2. Anti-inflammatory efficacy test

1) Cell inoculation: by 2.0X 10⁵Cell/well seeding Density cells were plated onto 24-well plates, incubators (37 ℃, 5% CO)₂) And incubated overnight.

2) Preparing a liquid: the test article working solution was prepared according to the experimental design (table 7).

TABLE 7 Experimental design

3) Administration: according to the experimental design shown in Table 7, when the cell plating rate in the 24-well plate reaches 40% -60%, the administration amount in each well is 1.0mL, each group is provided with 3 multiple wells, and the incubator (37 ℃ and 5% CO)₂) The cultivation was continued for 2 h.

4) Stimulation by LPS: after 2h of incubation, 100 μ L of LPS working solution prepared from the corresponding test substance working solution was added to the administered well plate according to the experimental design, the well plate was shaken from side to mix the drug in the well plate to a final concentration of 1 μ g/mL LPS, and the incubator (37 ℃ C., 5% CO)₂) The cultivation was continued for 22 h.

5) Collecting cells: after incubation and culture for 24h, 0.5mL of lysate was added to each well after the culture was completed, the mixture was left at room temperature for 5min to be sufficiently lysed, and the lysate was transferred to a 1.5mL RNase-free Eppendorf tube to extract RNA according to the protocol.

6) Reverse transcription: cDNA was synthesized according to the instructions of the RNA reverse transcription kit product.

7) qRT-PCR detection of iNOS gene was performed.

Third, statistical analysis of results

The graph Padprism Program software is used for drawing, t-test statistical analysis is adopted among groups, p < 0.05 indicates that the difference is obvious, and p < 0.01 indicates that the difference is extremely obvious.

Fourth, experimental results

1. Results of cytotoxicity assay

8 administration concentrations of the samples are set, cell viability tests are carried out by using Raw264.7 cells, the original data are shown in a table 8, the cell viability change trend is shown in a figure 6, and when the concentration of the sample LuAnna ice-water storage essence gel is less than or equal to 2.5% (V/V), the sample Luw264.7 cells are not cytotoxic.

Table 8 sample Lu Anna ice-moisture water-storage essence gel cytotoxicity test results

iNOS Gene expression test results

According to a specific experimental method, the content of iNOS gene was detected, the detection results are shown in Table 9, and the relative expression amounts are shown in FIG. 7, which are as follows:

TABLE 9 summary of iNOS Gene expression data

Sample name	Relative expression amount	SD	p-value
				BC	1.07	0.50	/
NC	11.53	1.63	0.005##
				PC	1.67	0.71	0.003**
Lu Anna ice-moistening water-storage essence gel-0.63%	2.04	0.77	0.004**
				Lu Anna ice-moistening water-storage essence gel-0.31%	2.54	0.62	0.004**
Lu Anna ice-moistening water-storage essence gel-0.16%	6.77	1.76	0.007**

Remarking: when the statistical analysis is carried out by the t-test method, the significance of the NC group is represented by # compared with the BC group, p-value < 0.05 is represented by #, and p-value < 0.01 is represented by # #. The significance of the sample group and PC group was indicated by p-value < 0.05 and p-value < 0.01, respectively, compared to the NC group.

Compared with the BC group, the iNOS expression level of the macrophage in the NC group is obviously increased, which shows that the LPS stimulation condition in the experiment is effective.

Compared with the NC group, the iNOS gene expression level of the macrophage is obviously reduced under the administration concentration of 100 mu g/mL of dexamethasone in the PC group, which shows that the experiment is effective.

Compared with an NC group, when the dosage concentration of the sample Lu' an ice-moistening water-storage essence gel is 0.63%, 0.31% and 0.16%, the iNOS gene expression level of macrophages is remarkably reduced.

In conclusion, when the dosage concentrations of the LuAnna ice-moistening water-storage essence gel are 0.63%, 0.31% and 0.16%, the iNOS gene expression level of macrophages is remarkably reduced, and the LuAnna ice-moistening water-storage essence gel is remarkably different from an NC group and has a relieving effect.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. The gel with the relieving effect is characterized in that the components of the gel comprise plant extracts, water, a humectant, a thickening agent and a preservative;

2. The jelly according to claim 1, wherein the mass of the plant extract is 0.05-6.1% of the mass of the jelly;

3. The gel according to claim 1, wherein the moisturizer is selected from one or more of glycerin, propylene glycol, sodium hyaluronate, glyceryl polyether-26 and methyl glucose polyether-20;

4. The jelly according to claim 1 or 3, wherein the humectant accounts for 0.05 to 23.1% by weight of the jelly.

5. The jelly according to claim 1, wherein the thickener is one or two selected from xanthan gum and sodium polyacrylate;

6. The method for preparing jelly according to any one of claims 1 to 5, which comprises the following steps: mixing water, humectant and thickener, and stirring to obtain a first stirred material;

7. The preparation method according to claim 6, wherein the stirring frequency of the first stirring is 35Hz, and the stirring time is 80 min;

8. The method for evaluating the soothing effect of the jelly is characterized by comprising the following steps of: by using an inflammatory cell model and taking iNOS expression quantity as an index, when the iNOS expression quantity is obviously lower than that of a negative control after gel is applied, the gel has the effect of relieving.

9. The method of claim 8, wherein the model of inflammatory cells comprises a model of inflammation established using lipopolysaccharide-stimulated macrophage raw264.7.

10. The method of claim 8, wherein the index further includes inflammatory state of body and change in skin moisture.