External dressing composition for skin

Abstract

The invention discloses an external dressing composition for skin, and relates to an external composition for skin. The skin external dressing composition is composed of corn stigma extract and radix ophiopogonis extract micro-sponge according to a weight part ratio of 1: 1. The skin barrier not only can lock skin moisture and grease, but also can resist invasion of various skin surface germs, and plays a very important role in protecting human health. The water content of normal skin is 10%-20%, and when the water content of the skin is reduced to below 10%, the skin barrier is damaged to a certain extent. The composition provided by the invention can promote differentiation and proliferation of keratinocytes so as to reduce inflammatory response, and also has an efficient moisturizing effect so as to improve skin functions. The invention solves the problems of dryness, desquamation, flushing, inflammation and the like after the skin barrier is damaged.

Description

Technical Field

[0001] The present invention relates to a composition for external use on skin. Background Art

[0002] The skin is the largest organ in the body, providing protection against physical, chemical and biological influences from the outside world. It also has various functions in regulating sensation, temperature regulation, secretion, excretion and immunity, and is the first barrier against mechanical damage and invading bacteria. At the same time, due to its direct contact with the outside world, the skin is one of the most vulnerable tissues. The skin barrier in a broad sense includes physical barriers, pigment barriers, neural barriers and immune barriers; in a narrow sense, the skin barrier refers to the epidermal permeability barrier, which is closely linked to other barriers and together constitutes the overall defense system of the skin barrier. Abnormalities in the function of one barrier will affect the function of other barriers. Large areas of skin exposed to the external environment may be mechanically damaged and invaded by bacteria. Once the skin tissue is damaged, the repair process is a complex collaborative process. Therefore, it is of great significance in the prevention and treatment of skin diseases and the development of daily cosmetics to regulate the integrity and function of the skin barrier and prevent and treat various skin problems caused by barrier damage. Summary of the invention

[0003] The invention provides a skin external dressing composition.

[0004] The skin external dressing composition of the present invention is composed of corn silk extract and ophiopogon japonicus extract micro-sponges in a weight ratio of 1:1.

[0005] The preparation method of the corn silk extract:

[0006] 1. The crushed dry corn silk is mixed with water to form a suspension, sterilized twice, and then inoculated with a mixed bacteria of Bifidobacterium adolescentis and yeast for fermentation to obtain a fermentation product;

[0007] 2. Add 70% ethanol to the fermented product for ultrasonic extraction, then filter, collect the filtrate, evaporate and concentrate, and vacuum freeze-dry to obtain corn silk extract.

[0008] The preparation method of the Ophiopogon japonicus extract micro-sponge:

[0009] 1. Preparation of Ophiopogon japonicus extract: add 80% ethanol to reflux extract of Ophiopogon japonicus powder, combine the extracts and filter, evaporate to remove ethanol until there is no alcohol taste, and freeze-dry to obtain Ophiopogon japonicus extract;

[0010] 2. Dissolve Ophiopogon japonicus extract and ethyl cellulose in dichloromethane to obtain phase A, mix acetone and water to prepare xanthan gum Tween 80 solution; disperse the xanthan gum Tween 80 solution in phase A, and then homogenize with an ultrasonic cell homogenizer until it becomes a uniform emulsion to obtain inner phase B; add water to PVA to swell completely as the outer phase, add inner phase B dropwise into the outer phase and stir until it is evenly dispersed, then evaporate the dichloromethane, filter, wash and dry to obtain the Ophiopogon japonicus extract micro sponge.

[0011] Beneficial effects of the present invention:

[0012] First, (1) corn silk extract has good antioxidant effect. Antioxidant experiment was conducted to examine the effect of extracts from each group on scavenging DPPH free radicals. Antioxidant can protect the skin from free radical damage. Antioxidant can enhance skin resistance and anti-photoaging ability. Antioxidant can resist inflammation and reduce the occurrence of problems such as spots and sensitivity. (2) HaCaT cells undergo abnormal proliferation after being treated with sodium dodecyl sulfate (SLS). After being given the composition, the proliferation of cells can be inhibited. It is shown that the composition was used to perform skin repair cell experiments. (3) A mouse skin barrier damage model was established by the tape method. After being given the composition, ceramide in the skin tissue was detected. After combined intervention, the ceramide content increased. The composition can replenish the natural moisturizing factor ceramide in the skin. Ceramide, as a key component of intercellular lipids in the skin stratum corneum, is essential for maintaining the skin barrier function. It mainly strengthens the connection between keratinocytes to form a waterproof barrier, effectively prevents water loss, and improves the skin's water retention. Ceramide can promote cell growth, increase the thickness of the stratum corneum, and enhance the skin's resistance to external stimuli. (4) After HE staining and microscopic examination, it was found that the skin tissue structure of the model group was moderately abnormal, the epidermal structure was missing in the visual field, and a small amount of inflammatory cell infiltration was seen in the tissue, confirming that the skin was damaged and the skin barrier was damaged. After administration to the medium-dose group, the skin tissue structure was slightly abnormal, the epidermal structure was clear in the visual field, and no obvious inflammatory cell infiltration was seen in the tissue, confirming that it had a skin repair effect. (5) The ELISA method was used to determine the TNF-α content in the dorsal skin tissue. After intervention, the TNF-α content decreased to varying degrees, and the decrease in the medium-dose group was the most significant, proving that it has an anti-inflammatory effect.

[0013] The method for preparing corn silk extract of the present invention adopts microbial fermentation, which is a biotransformation method. The effective components of Chinese herbal medicine after microbial fermentation can be fully extracted and better exert their efficacy. Yeast, as a natural fermentation agent, produces intracellular enzymes and extracellular enzymes during its own growth and metabolism, which can increase the dissolution rate of polyphenols and improve the biological activity of polyphenols. Corn silk contains high fiber substances. Under the action of composite microorganisms, the cell wall of cellulose is transformed by yeast and youth.

[0014] The enzyme metabolism of bifidobacteria promotes the release of polyphenols. Therefore, after microbial fermentation, the polyphenol content in corn silk increases.

[0015] The preparation of the Ophiopogon japonicus extract microsponge of the present invention adopts the improved technology of xanthan gum-assisted ethyl cellulose sponge, and uses the W / O / W type emulsification method to prepare the xanthan gum-solubilized Ophiopogon japonicus extract-ethyl cellulose microsponge. The microsponge is a porous polymer microcarrier with a large number of pores on the surface. Its porosity increases the surface area to volume ratio, can carry drugs several times the weight of the matrix, and control the release rate of the encapsulated drug. The preparation of the Ophiopogon japonicus extract into a microsponge preparation can overcome the limitations of other microcarriers, such as low drug loading, instability, slow drug release rate, etc., and improve the effect of local administration.

[0016] The skin dressing composition of the present invention uses corn silk extract and ophiopogon japonicus microsponges in a ratio of 1:1 as core ingredients for the preparation of a skin dressing composition. The skin dressing composition is preferably a cosmetic composition, such as a face cream, essence, emulsion, etc., and the preferred weight percentage of the composition is 0.01%-10% (w / w).

[0017] The skin barrier can not only lock in skin moisture and oil, but also resist the invasion of various skin surface pathogens, and plays a very important protective role in human health. The normal skin water content is 10% to 20%. When the skin water content drops below 10%, the skin barrier will be damaged to a certain extent. The composition of the present invention can promote the differentiation and proliferation of keratinocytes, thereby reducing inflammatory reactions, and at the same time has a high-efficiency moisturizing effect and improves skin function. The present invention solves the problems of dryness, desquamation, flushing, inflammation, etc. that occur after the skin barrier is damaged. The composition of the present invention can supplement the natural moisturizing factor ceramide in the skin. Ceramide, as a key component of the intercellular lipids in the stratum corneum of the skin, is essential to maintaining the skin barrier function. It mainly strengthens the connection between keratinocytes to form a waterproof barrier, effectively prevents water loss, and improves the water retention of the skin. Ceramide can promote cell growth, increase the thickness of the stratum corneum, and enhance the skin's resistance to external stimuli. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the amount of polyphenol extraction;

[0019] Figure 2 The DPPH free radical scavenging ability was determined by spectrophotometry;

[0020] Figure 3 The flowchart of the preparation of Ophiopogon japonicus extract microsponges;

[0021] Figure 4 It is freeze-dried powder of Ophiopogon japonicus extract;

[0022] Figure 5 It is a micro sponge containing Ophiopogon japonicus extract;

[0023] Figure 6 This is a transmission electron microscopic image of the microsponge of Ophiopogon japonicus extract;

[0024] Figure 7 is the in vitro release curve;

[0025] Figure 8 is the DPPH free radical scavenging rate;

[0026] Fig. 9 This is the result graph of cell proliferation rate;

[0027] Fig.10 HE staining microscopy results of the blank group;

[0028] Fig.11 HE staining microscopy results of the model group;

[0029] Fig.12 HE staining microscopic examination results of the vehicle group;

[0030] Fig.13 This is the result of HE staining microscopy in the low-dose group;

[0031] Fig.14 HE staining microscopy results of the medium-dose group;

[0032] Fig.15 HE staining microscopy results of the high-dose group;

[0033] Fig.16 It is the result diagram of the determination of Cer content;

[0034] Fig.17 This is a graph showing the results of measuring TNF-α content. DETAILED DESCRIPTION

[0035] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0037] Specific implementation method 1: The skin external dressing composition of this implementation method is composed of corn silk extract and ophiopogon japonicus extract microsponges in a weight ratio of 1:1.

[0038] Specific embodiment 2: This embodiment is different from the specific embodiment 1 in that: the preparation method of the corn silk extract is:

[0039] 1. The crushed dry corn silk is mixed with water to form a suspension, sterilized twice, and then inoculated with a mixed bacteria of Bifidobacterium adolescentis and yeast for fermentation to obtain a fermentation product;

[0040] 2. Add 70% ethanol to the fermented product for ultrasonic extraction, then filter, collect the filtrate, evaporate and concentrate, and vacuum freeze-dry to obtain corn silk extract. Other steps and parameters are the same as those in the first embodiment.

[0041] In step 1 of this embodiment, the dry corn silk is obtained by drying the corn silk in the shade.

[0042] Specific implementation method 3: This implementation method is different from specific implementation method 2 in that the material-liquid ratio of the crushed dry corn silk to water in step 1 is 60 g:40 L. Other steps and parameters are the same as those in specific implementation method 2.

[0043] Specific embodiment 4: This embodiment differs from specific embodiment 2 in that the inoculation amount in step 1 is a mixture of 5% Bifidobacterium adolescentis fermentation broth and 5% yeast fermentation broth, and the ratio of the number of active bacteria in Bifidobacterium adolescentis fermentation broth to that in yeast fermentation broth is 1:1. Other steps and parameters are the same as those in specific embodiment 2.

[0044] In this embodiment, the bacterial cell concentration in the Bifidobacterium adolescentis fermentation liquid and the yeast fermentation liquid is 5%.

[0045] Specific embodiment 5: This embodiment differs from specific embodiment 2 in that the material-liquid ratio of the fermentation product to 70% ethanol in step 2 is 1:12 g / mL, the ultrasonic extraction temperature is 50°C, and the ultrasonic time is 30 min. Other steps and parameters are the same as those in specific embodiment 2.

[0046] Specific embodiment 6: This embodiment is different from the specific embodiment 1 in that: the preparation method of the Ophiopogon japonicus extract microsponge is:

[0047] 1. Preparation of Ophiopogon japonicus extract: add 80% ethanol to reflux extract of Ophiopogon japonicus powder, combine the extracts and filter, evaporate to remove ethanol until there is no alcohol taste, and freeze-dry to obtain Ophiopogon japonicus extract;

[0048] 2. Dissolve Ophiopogon japonicus extract and ethyl cellulose in dichloromethane to obtain phase A, mix acetone and water to prepare xanthan gum Tween 80 solution; disperse the xanthan gum Tween 80 solution in phase A, and then homogenize with an ultrasonic cell homogenizer until it becomes a uniform emulsion to obtain inner phase B; add water to PVA to swell completely as the outer phase, add inner phase B dropwise into the outer phase and stir until it is evenly dispersed, then evaporate the dichloromethane, filter, wash and dry to obtain the Ophiopogon japonicus extract micro sponge.

[0049] In step 1 of this embodiment, Ophiopogon japonicus is taken, crushed and passed through a 40-mesh sieve to obtain Ophiopogon japonicus powder.

[0050] In step 2 of this embodiment, the inner phase B is slowly added dropwise into the outer phase, and a magnetic stirrer is used to continuously stir until the phases are uniformly dispersed; the inner phase B is slowly added dropwise into the outer phase in order to allow the inner phase B and the outer phase to react more completely.

[0051] Specific embodiment 7: This embodiment is different from specific embodiment 6 in that: in step 1, 8 times 80% ethanol is added to the ophiopogon powder for reflux extraction twice, each extraction is for 4 hours. Other steps and parameters are the same as those of specific embodiment 6.

[0052] Specific embodiment eight: This embodiment differs from specific embodiment six in that the volume ratio of acetone to water in step two is 1:5, the concentration of xanthan gum in the xanthan gum Tween 80 solution is 0.1%, and the concentration of Tween 80 solution is 0.1%. Other steps and parameters are the same as those in specific embodiment six.

[0053] Specific embodiment 9: This embodiment is different from specific embodiment 6 in that: in step 2, the xanthan gum Tween 80 solution is dispersed in phase A, and the ultrasonic cell homogenizer is operated at a power of 100 W for 3 seconds and then rests for 3 seconds, and continuously homogenized until it becomes a uniform emulsion. Other steps and parameters are the same as those of specific embodiment 6.

[0054] Specific embodiment 10: This embodiment differs from specific embodiment 6 in that: in step 2, the dichloromethane is evaporated, filtered, washed, and dried, that is, after the dichloromethane is evaporated in a fume hood, it is filtered, washed twice with distilled water, and dried in an oven at 60°C. The other steps and parameters are the same as those of specific embodiment 6.

[0055] Example 1 Comparative experiment on the preparation of corn silk extract

[0056] Experimental groups:

[0057] Group A: corn silk was inoculated with Bifidobacterium adolescentis only;

[0058] Group B: corn silk inoculated with yeast only;

[0059] Group C: corn silk was first inoculated with yeast and then with Bifidobacterium adolescentis;

[0060] Group D: corn silk was first inoculated with Bifidobacterium adolescentis and then with yeast;

[0061] Group E: corn silk was inoculated with Bifidobacterium adolescentis and yeast simultaneously;

[0062] Group F: unfermented corn silk.

[0063] Method for activating Bifidobacterium adolescentis strain: place an ampoule containing Bifidobacterium adolescentis (strain CICC 24573 Bifidobacterium genus) in a biosafety cabinet, disinfect it with an alcohol cotton ball, take 1 ml of MRS liquid culture medium and put it in the ampoule, shake and mix, inoculate the bacterial suspension into MRS liquid culture medium at an inoculum size of 5%, and after anaerobically culturing at 37°C for 18 hours, take the bacterial suspension and inoculate three zones into MRS solid culture medium to purify the strain, and anaerobically culture it at 37°C for 24 hours, pick a single colony and culture it in MRS liquid culture medium for 24 hours to obtain Bifidobacterium adolescentis fermentation liquid.

[0064] Yeast activation method: Yeast extract peptone glucose medium (YEPD): Weigh 2g of yeast powder, 2g of peptone, and 2g of glucose in a 250mL conical flask, add 100mL of pure water, and sterilize at 121℃ for 25min. After cooling to room temperature, store in a refrigerator at 4℃.

[0065] Yeast extract peptone dextrose agar medium (YPD): Weigh 1g yeast powder, 2g peptone, 2g glucose, and 2g agar powder in a 250mL conical flask, add 100mL pure water, and sterilize at 121℃ for 25min. Cool to 50-60℃, pour into a sterile culture dish, and store in a 4℃ refrigerator after solidification.

[0066] Yeast powder was mixed with PBS in a certain proportion, and after reaction for 30 minutes, it was inoculated into YPD using a bacterial loop for screening and purification. Single colonies in the solid culture medium were picked and inoculated into YEPD for propagation. The culture was carried out at 30°C and 180 r / min for 8 hours to obtain yeast liquid.

[0067] Preparation of fermentation products of each group:

[0068] Group A: Weigh an appropriate amount of corn silk, dry it in the shade, grind it through a 40-mesh sieve, and place it in a 250-mL conical flask. Prepare a suspension with water at a solid-liquid ratio of 60 g:40 L, sterilize it under high pressure at 121°C for 25 min, cool it to room temperature, and sterilize it under ultraviolet light in a biosafety cabinet for 30 min. Inoculate the suspension with Bifidobacterium adolescentis fermentation liquid at a 5% inoculation rate, and culture it in a constant temperature shaker for 3 days to obtain Bifidobacterium adolescentis fermentation product.

[0069] Group B: Weigh an appropriate amount of corn silk, dry it in the shade, grind it through a 40-mesh sieve, and place it in a 250-mL conical flask. Prepare a suspension with water at a solid-liquid ratio of 60 g:40 L, sterilize it under high pressure at 121°C for 25 min, cool it to room temperature, and sterilize it under ultraviolet light in a biosafety cabinet for 30 min. Inoculate the yeast fermentation liquid with an inoculum size of 5% and culture it in a constant temperature shaker for 3 days to obtain yeast fermentation product.

[0070] Group C: The steps were the same as those for corn silk treatment in Group B, except that 5% yeast fermentation liquid was inoculated first, and then 5% Bifidobacterium adolescentis fermentation liquid was inoculated after 3 days of fermentation.

[0071] Group D: The steps were the same as those for corn silk treatment in Group A, except that 5% Bifidobacterium adolescentis fermentation broth was first inoculated, and then 5% yeast fermentation broth was inoculated after 3 days of fermentation.

[0072] Group E (this embodiment simultaneously inoculates Bifidobacterium adolescentis and yeast): Weigh an appropriate amount of corn silk, dry it in the shade, grind it through a 40-mesh sieve, and place it in a 250mL conical flask. Prepare a suspension of the crushed corn silk with water at a solid-liquid ratio of 60g:40L, sterilize it under high pressure at 121°C for 25min, cool it to room temperature, and sterilize it under ultraviolet light in a biosafety cabinet for 30min. Inoculate a mixed bacteria of 5% Bifidobacterium adolescentis fermentation broth (with a bacterial concentration of 5%) and 5% yeast fermentation broth (with a bacterial concentration of 5%) at a concentration of 1:1, and obtain a fermentation product after fermentation for 3 days.

[0073] Group F: Weigh an appropriate amount of corn silk, dry it in the shade, and grind it through a 40-mesh sieve.

[0074] The corn silk extract is prepared by using the above fermentation products, and the specific method is as follows:

[0075] The obtained materials of A, B, C, D, E and F were placed in a 250mL conical flask, and 70% ethanol was added, with a solid-liquid ratio of 1:12 (g / mL), ultrasonic extraction temperature of 50°C, ultrasonic time of 30min, and then filtered after completion. The filtrate was collected and concentrated by evaporation, and vacuum freeze-dried for later use to obtain the extract of corn silk fermentation (except group F).

[0076] The fermented corn silk extracts in each group were tested:

[0077] The obtained materials of A, B, C, D, E and F were placed in a 250mL conical flask, and 70% ethanol was added, with a solid-liquid ratio of 1:12 (g / mL), ultrasonic extraction temperature of 50°C, ultrasonic time of 30min, and then filtered after completion. The filtrate was collected and concentrated by evaporation, and vacuum freeze-dried for later use to obtain the extract of corn silk fermentation (except group F).

[0078] 1. Determination of total phenol content

[0079] Preparation of gallic acid standard solution: Accurately weigh 25.00 mg of gallic acid that has been fully dried to a constant weight and add it to 25 mL of deionized water to obtain a 1 mg / mL gallic acid solution. Pipette 5 mL of the 1 mg / mL gallic acid solution and dilute to a 50 mL volumetric flask with deionized water. Shake well to obtain a 0.1 mg / mL gallic acid solution.

[0080] Drawing of standard curve: Take the above gallic acid standard solution, then add 0.5mL of Folin phenol reagent and 2mL of 10% sodium carbonate solution respectively, make the volume to 10mL, react at room temperature for 2h, measure the absorbance (y) at 760nm with UV-visible spectrophotometer, x is gallic acid concentration (μg / mL). The standard curve equation is: y=0.2583x+0.0547, correlation coefficient R2=0.9951.

[0081] The polyphenol content was determined according to the above method, and the polyphenol content and extraction rate were calculated by the standard curve equation: polyphenol extraction amount = mass of polyphenols in the extract (g) / mass of the raw material (g) × 100%

[0082] Figure 1 The polyphenol extraction results of each group of experiments are shown in Table 1. Figure 1 It can be seen that after the E group (in this embodiment, Bifidobacterium adolescentis and yeast were inoculated at the same time) was fermented and extracted, and the polyphenol extraction amount was detected after extraction. It can be seen that the polyphenol content of the E group was the highest, indicating that after the fermentation of corn silk, it is helpful to increase the polyphenol content. In addition, the polyphenol content of the corn silk extracted after the composite fermentation was higher than the polyphenol content of the corn silk extracted after the single bacterial fermentation, and was much higher than the polyphenol content of the non-fermented corn silk extract.

[0083] Microbial fermentation is a method of biological transformation. Studies have found that the effective ingredients of Chinese herbal medicines can be fully extracted after microbial fermentation and better exert their efficacy. Yeast, as a natural fermentation agent, produces intracellular and extracellular enzymes during its own growth and metabolism, which can increase the dissolution rate of polyphenols and improve the biological activity of polyphenols. Corn silk contains a high amount of fiber. Under the action of composite microbial bacteria, the cell wall of cellulose is metabolized by the enzyme system of yeast and Bifidobacterium adolescentis degrading bacteria, which promotes the release of polyphenols. Therefore, after microbial fermentation, the polyphenol content in corn silk increases.

[0084] 2. Antioxidant test

[0085] The extracts obtained from different groups were prepared into solutions with mass concentrations of 20, 25, 30, 35 and 40 μg / mL for antioxidant tests. The effects of the extracts from each group on scavenging DPPH free radicals were investigated. DPPH is a stable free radical with a maximum absorption peak at 517nm. When the test solution contains a free radical scavenger, it will capture the unpaired electrons of DPPH, causing the color to change to yellow. The smaller the absorbance, the stronger the scavenging ability.

[0086] Figure 2 The DPPH free radical scavenging ability was determined by spectrophotometry. Figure 2 It can be seen that after extracting polyphenols from group E, i.e. compound fermented corn silk, the DPPH free radical scavenging rate reached 92.1% at a concentration of 40ug / ml, indicating good antioxidant properties, which is much higher than other extraction methods.

[0087] Example 2 Comparative experiment on preparation of micro-sponge with Ophiopogon japonicus extract

[0088] Ophiopogon microsponge part:

[0089] Microsponge is a porous polymer microcarrier with a large number of pores on its surface. Its porosity increases the surface area to volume ratio, and it can carry drugs several times the weight of the matrix and control the release rate of the encapsulated drugs. The preparation of Ophiopogon japonicus extract into microsponge preparations can overcome the limitations of other microcarriers, such as low drug loading, instability, slow drug release, etc., and improve the effect of local drug delivery.

[0090] The invention adopts the improved technology of xanthan gum-assisted ethyl cellulose sponge and uses W / O / W type emulsification method to prepare xanthan gum-solubilized ophiopogon japonicus extract-ethyl cellulose micro-sponge.

[0091] Preparation process of Ophiopogon japonicus extract microsponge:

[0092] Preparation of Ophiopogon japonicus extract: Take a certain amount of Ophiopogon japonicus, crush it and pass it through a 40-mesh sieve. Add 8 times the amount of 80% ethanol and reflux and extract twice, each time for 4 hours, combine the extracts and filter, evaporate the ethanol until there is no alcohol taste, and freeze-dry to obtain the Ophiopogon japonicus extract.

[0093] Microsponge preparation process:

[0094] Take an appropriate amount of Ophiopogon japonicus extract and ethyl cellulose and dissolve them in dichloromethane as phase A, and use acetone: water (1:5) as solvent to prepare xanthan gum solution. Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100W, work for 3s and then stand for 3s, and continuously homogenize until it is a uniform emulsion, which is the inner phase B. Take an appropriate amount of PVA in a beaker, add water to swell completely, as the outer phase. Slowly add the inner phase B dropwise into the outer phase, and use a magnetic stirrer to continue stirring until it is evenly dispersed. Place the resulting mixture in a fume hood to evaporate the dichloromethane, filter it, wash it twice with distilled water, and dry it in a 60°C oven.

[0095] 1. Set up 9 groups of experiments. The specific methods are as follows:

[0096] Experimental Group 1:

[0097] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (3:1) and dissolve them in dichloromethane to prepare phase A.

[0098] Preparation of internal phase B: Prepare 0.2% xanthan gum solution with acetone: water (1:5) as solvent. Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0099] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 1.0% as the external phase.

[0100] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0101] Experimental Group 2:

[0102] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0103] Preparation of internal phase B: Prepare 0.2% xanthan gum solution with acetone: water (1:5) as solvent. Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0104] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 1.0% as the external phase.

[0105] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0106] Experimental Group 3:

[0107] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (7:1) and dissolve them in dichloromethane as phase A;

[0108] Preparation of internal phase B: Prepare 0.2% xanthan gum solution with acetone: water (1:5) as solvent. Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0109] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 1.0% as the external phase.

[0110] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0111] Experimental Group 4:

[0112] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0113] Preparation of internal phase B: Prepare 0.2% Tween 80 solution with acetone: water (1:5) as solvent. Take an appropriate amount of Tween 80 solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0114] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 1.0% as the external phase.

[0115] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0116] Experimental Group 5:

[0117] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0118] Preparation of internal phase B: Acetone: water (1:5) was used as solvent to prepare xanthan gum Tween 80 solution (0.1% xanthan gum + 0.1% Tween 80 solution, the method to obtain 0.1% xanthan gum Tween solution: 0.1g xanthan gum was added to 100g water, the concentration was 0.1%; 0.1% Tween was also added to 0.1g Tween to 100g water, the concentration was 0.1%), and the xanthan gum Tween 80 solution was dispersed in phase A. Take an appropriate amount of xanthan gum Tween 80 solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100W, work for 3s and then rest for 3s, and continuously homogenize until it is a uniform milky state, which is internal phase B.

[0119] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 1.0% as the external phase.

[0120] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0121] Experimental Group 6

[0122] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0123] Preparation of internal phase B: A xanthan gum Tween 80 solution (0.1% xanthan gum + 0.1% Tween 80 solution) was prepared using acetone: water (1:5) as solvent. An appropriate amount of xanthan gum Tween 80 solution was dispersed in phase A. An ultrasonic cell homogenizer was used at a power of 100 W, working for 3 seconds and then resting for 3 seconds, and continuous homogenization was performed until a uniform emulsion was formed, which was internal phase B.

[0124] Preparation of external phase: Take an appropriate amount of PVA in a beaker and add water to swell it completely to a concentration of 2.0% as the external phase.

[0125] Slowly drop the inner phase B into the outer phase and continue stirring with a magnetic stirrer until it is evenly dispersed. After the resulting mixture is placed in a fume hood to evaporate the dichloromethane, it is filtered and washed twice with distilled water, and then placed in an oven at 60°C to dry.

[0126] Experimental Group 7:

[0127] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0128] Preparation of internal phase B: Prepare xanthan gum Tween 80 solution (0.1% xanthan gum + 0.1% Tween 80 solution) using acetone: water (1:5) as solvent. Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0129] Preparation of the external phase: Take an appropriate amount of PVA in a beaker, add water to swell completely, and make its concentration 0.5% as the external phase. Slowly drop the internal phase B into the external phase, and use a magnetic stirrer to continuously stir until it is evenly dispersed. Place the resulting mixture in a fume hood to evaporate the dichloromethane, filter it, wash it with distilled water twice, and dry it in a 60°C oven.

[0130] Experimental Group 8:

[0131] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0132] Preparation of internal phase B: Acetone: water (1:5) was used as solvent to prepare (1.0% concentration of xanthan gum). An appropriate amount of xanthan gum solution was dispersed in phase A. An ultrasonic cell homogenizer was used with a power of 100 W, working for 3 seconds and then resting for 3 seconds, and continuous homogenization was performed until a uniform emulsion was formed, which was internal phase B.

[0133] Preparation of external phase: Take an appropriate amount of PVA in a beaker, add water to swell completely, and make its concentration 1.0% as the external phase. Slowly drop the internal phase B into the external phase, and use a magnetic stirrer to continuously stir until it is evenly dispersed. Place the resulting mixture in a fume hood to evaporate the dichloromethane, filter it, wash it with distilled water twice, and dry it in a 60°C oven.

[0134] Experimental Group 9:

[0135] Preparation of phase A: Take appropriate amount of Ophiopogon japonicus extract and ethyl cellulose (5:1) and dissolve them in dichloromethane as phase A;

[0136] Preparation of internal phase B: Prepare 1.0% Tween 80 with acetone: water (1:5) as solvent. Disperse an appropriate amount of xanthan gum solution in phase A. Use an ultrasonic cell homogenizer with a power of 100 W, work for 3 seconds and then rest for 3 seconds, and continuously homogenize until it becomes a uniform emulsion, which is internal phase B.

[0137] Preparation of external phase: Take an appropriate amount of PVA in a beaker, add water to swell completely, and make its concentration 1.0% as the external phase. Slowly drop the internal phase B into the external phase, and use a magnetic stirrer to continuously stir until it is evenly dispersed. Place the resulting mixture in a fume hood to evaporate the dichloromethane, filter it, wash it with distilled water twice, and dry it in a 60°C oven.

[0138] 2. Testing the above 9 experimental groups

[0139] 1. Determination of encapsulation efficiency and content of ruscoside in Ophiopogon japonicus extract microsponges

[0140] Preparation of reference solution and test solution: accurately weigh 5 mg of ruscoside aglycone reference, add appropriate amount of methanol and ultrasonicate for 5 min to dissolve, then dilute to 100 mL to prepare 50 μg / mL reference solution. Accurately weigh 100 mg of Ophiopogon japonicus extract microsponges, add appropriate amount of methanol and ultrasonicate for 5 min to dissolve, then dilute to 100 mL, and shake well to obtain the test solution.

[0141] Linear relationship investigation: Referring to the method in the "Chinese Pharmacopoeia" (2020 edition), accurately measure 0.5mL, 1mL, 2mL, 3mL, 4mL, 5mL, and 6mL of the reference solution, respectively, and place them in stoppered test tubes, evaporate the solvent in a water bath, accurately add 10mL of perchloric acid, shake well, keep warm in 80℃ hot water for 15 minutes, take out, cool with ice water, use the corresponding reagent as blank, and use UV-visible spectrophotometry (General Rule 0401) to measure the absorbance A at a wavelength of 397nm. With A as the abscissa and the mass concentration of the reference solution (Y) as the ordinate, linear regression is performed to obtain the equation Y=478.61X+17.786, R 2 =0.9943 (n=6), indicating that the content of ruscosapogenin standard in the range of 25-300 μg has a good linear relationship with the absorbance.

[0142] Encapsulation efficiency = (m act / m the )×100%

[0143] Where m act is the mass of ruscoside in microsponges measured by UV-spectrophotometry, m the It is the mass of ruscoside aglycone contained in the freeze-dried powder of Ophiopogon japonicus extract of the same mass.

[0144] The encapsulation efficiency results of the 9 experimental groups are shown in Table 1.

[0145] Table 1 Encapsulation efficiency results

[0146]

[0147]

[0148] As can be seen from Table 1, the encapsulation efficiency of the drug-ethyl cellulose (experimental group 1) was 17.01% when it was close to 3:1. The encapsulation efficiency of the drug-ethyl cellulose (experimental group 2) increased significantly when it was close to 5:1. When the content of the Ophiopogon japonicus extract continued to increase (experimental group 3) to 7:1, the encapsulation efficiency would suddenly decrease. It is speculated that ethyl cellulose, as the skeleton structure material of the microsponge, is difficult to completely encapsulate when the drug content is too high, resulting in the leakage of the main component to the outer phase, leading to a significant decrease in the encapsulation efficiency. Mixing Tween and xanthan gum (experimental group 5) plays a role in enhancing the stability of the system. In experimental group 8, when the concentration of xanthan gum was 1.0%, and in experimental group 9, when the concentration of Tween was 1.0%, the microsponge morphology could not be formed. This may be due to the high viscosity of the system, which is not conducive to the volatilization of dichloromethane, and a complete and dry microsponge morphology cannot be obtained. In experimental group 6, when the concentration of PVA was 2.0%, the microsponge state could not be formed. It is speculated that this may be because the concentration of PVA is too high, and the internal phase is over-solidified by PVA, resulting in the formation of micropores, but the loaded drug cannot be released into the medium through the micropores.

[0149] The encapsulation efficiency of experimental group 5 was the highest, and the drug was encapsulated most completely. Mixing Tween and xanthan gum (experimental group 5) plays a role in enhancing the stability of the system.

[0150] Detection of the Ophiopogon japonicus extract microsponge in Example 3

[0151] Preparation of the Ophiopogon japonicus extract: Take a certain amount of Ophiopogon japonicus, crush it and sieve it through a 40-mesh sieve. Add 8 times the amount of 80% ethanol and reflux extract it twice, 4 hours each time. Combine the extractive solutions, filter them, and evaporate the ethanol to no alcohol smell by rotary evaporation, and then freeze-dry to obtain the Ophiopogon japonicus extract.

[0152] Preparation of the Ophiopogon japonicus extract microsponge ( Figure 3 Flow chart for the preparation of the Ophiopogon japonicus extract microsponge) The method is as follows:

[0153] Take an appropriate amount of the Ophiopogon japonicus extract and ethyl cellulose and dissolve them in dichloromethane as phase A. Prepare a xanthan gum solution with acetone: water (1:5) as the solvent. Take an appropriate amount of the xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer at a power of 100 W, work for 3 s and then stand still for 3 s, and continuously homogenize until it becomes a uniform emulsion, which is the internal phase B. Take an appropriate amount of PVA in a beaker, add water and swell it completely as the outer phase. Slowly drop the internal phase B into the outer phase, and continuously stir with a magnetic stirrer until it is uniformly dispersed. Place the obtained mixture in a fume hood to evaporate dichloromethane, then filter it, wash it twice with distilled water, and dry it in an oven at 60 °C to obtain it.

[0154] 1. The morphology and appearance of the Ophiopogon japonicus extract in this example are as Figures 4 to 6 shown, where Figure 4 is the freeze-dried powder of the Ophiopogon japonicus extract, Figure 5 is the Ophiopogon japonicus extract microsponge, Figure 6 This is a transmission electron microscopic image of the microsponge of Ophiopogon japonicus extract. Figure 4 and Figure 5 From the appearance, it can be seen that compared with the freeze-dried powder of Ophiopogon japonicus extract, the appearance of the microsponge is white to light yellow before crushing, and the surface is loose and porous. Figure 6 The microsponges were observed to be porous spheres under transmission electron microscopy, and were evenly distributed under scanning electron microscopy, showing a porous spherical structure.

[0155] 2. Release test of the microsponges of Ophiopogon japonicus extract in this example

[0156] Boil the dialysis bag for 10 minutes to activate it, then expel the air from the bag and clamp one end. Take appropriate amounts of Ophiopogon japonicus extract and Ophiopogon japonicus extract microsponges (microsponges), respectively, dissolve them in 10 mL of deionized water and place them in the dialysis bag. Clamp the other end. Add 200 mL of physiological saline solution containing 30% ethanol to the beaker as the release medium. Suspend the dialysis bag in the beaker and conduct the experiment at a speed of 800 r / min and 37°C. Take 1 mL of the release solution at 1, 2, 3, 4, 5, 6, 12, and 24 h, respectively, and fill it with an equal amount of release medium at the same temperature. Determine according to the method in 3.3. The in vitro release curve is shown in Figure 7 As shown, from Figure 7 It can be seen that the cumulative release of the API reached 92.67% within 6 hours and was almost completely released within 12 hours. The release rate of the microsponges was 79.64% within 12 hours and 90.85% after 24 hours, indicating that the microsponges had a certain sustained release effect.

[0157] 3. Determination of free radical scavenging ability of Ophiopogon japonicus extract by DPPH method

[0158] Weigh a certain amount of DPPH and prepare a 0.04 mg / mL DPPH solution with anhydrous ethanol. Take 2 mL of different concentrations of Ophiopogon japonicus extract solution, add 2 mL of DPPH solution, mix well, leave at room temperature for 30 minutes, and centrifuge at 5000 r / min for 10 minutes. Take the supernatant and measure the absorbance at 515 nm. Use Vc as a positive control. The scavenging rate of the sample for DPPH free radicals is calculated using the following formula: DPPH free radical scavenging rate % = [[A 0 -(A 1 -A 2 )]÷A 0 ]×100%

[0159] A 0 —Absorbance value of 2mL anhydrous ethanol + 2mL DPPH solution;

[0160] A 1 —Absorbance value of 2mL sample solution + 2mL DPPH solution;

[0161] A 2 —The absorbance value of 2mL sample solution + 2mL anhydrous ethanol.

[0162] Figure 8 is the DPPH free radical scavenging rate, Figure 8 It can be seen that the free radical scavenging ability of Ophiopogon japonicus is not outstanding. Because of its weak ability to scavenge free radicals, it needs to be combined with corn silk extract, which has very strong antioxidant ability, so that multiple effects can be achieved.

[0163] Example 4 Testing of the composition with skin repairing effect obtained by the present invention

[0164] The skin external dressing composition of this embodiment is composed of corn silk extract and ophiopogon japonicus extract microsponges in a weight ratio of 1:1;

[0165] 1. The preparation method of the corn silk extract is as follows:

[0166] 1.1. Method for activating Bifidobacterium adolescentis strains:

[0167] Place an ampoule containing Bifidobacterium adolescentis (strain CICC 24573 Bifidobacterium genus) in a biosafety cabinet and disinfect it with an alcohol cotton ball. Take 1 ml of MRS liquid culture medium and put it in the ampoule. Shake and mix. Inoculate the bacterial suspension in the MRS liquid culture medium at an inoculum of 5%. After anaerobic culture at 37°C for 18 hours, take the bacterial suspension and inoculate three zones in MRS solid culture medium to purify the strain. After anaerobic culture at 37°C for 24 hours, pick a single colony and culture it in the MRS liquid culture medium for 24 hours to obtain the fermentation liquid of Bifidobacterium adolescentis.

[0168] 1.2. Yeast activation method:

[0169] Yeast extract peptone dextrose medium (YEPD): Weigh 2g of yeast powder, 2g of peptone, and 2g of glucose in a 250mL conical flask, add 100mL of pure water, and sterilize at 121℃ for 25min. After cooling to room temperature, store in a refrigerator at 4℃.

[0170] Yeast extract peptone dextrose agar medium (YPD): Weigh 1g of yeast powder, 2g of peptone, 2g of glucose, and 2g of agar powder in a 250mL conical flask, add 100mL of pure water, and sterilize at 121°C for 25min. Cool to 50-60°C, pour into a sterile culture dish, and store in a refrigerator at 4°C after solidification;

[0171] Yeast powder was mixed with PBS in a certain proportion, and after reaction for 30 minutes, it was inoculated into YPD using a bacterial loop for screening and purification. Single colonies in the solid culture medium were picked and inoculated into YEPD for propagation. The culture was carried out at 30°C and 180 r / min for 8 hours to obtain yeast liquid.

[0172] 1.3. Weigh an appropriate amount of corn silk, dry it in the shade, crush it and pass it through a 40-mesh sieve, place it in a 250mL conical flask, mix the crushed corn silk with water in a ratio of 60g:40L to form a suspension, sterilize it at high pressure at 121℃ for 25min, cool it to room temperature, sterilize it with ultraviolet light in a biosafety cabinet for 30min, and inoculate a mixed bacteria of 5% Bifidobacterium adolescentis fermentation broth and 5% yeast fermentation broth at a concentration of 1:1. After fermentation for 3 days, obtain the fermentation product;

[0173] 1.4. Fermented corn silk extract:

[0174] The fermented product obtained in step 3 was placed in a 250 mL conical flask, 70% ethanol was added, the solid-liquid ratio was 1:12 (g / mL), the ultrasonic extraction temperature was 50°C, the ultrasonic time was 30 min, and after the end, the filtrate was collected and evaporated and concentrated, and vacuum freeze-dried for standby use to obtain a corn silk fermented product extract;

[0175] 2. The preparation method of Ophiopogon japonicus extract microsponge is as follows:

[0176] 2.1. Preparation of Ophiopogon japonicus extract: Take a certain amount of Ophiopogon japonicus, crush it and pass it through a 40-mesh sieve. Add 8 times the amount of 80% ethanol and reflux and extract twice, each time for 4 hours, combine the extracts and filter, evaporate the ethanol by rotary evaporation until there is no alcohol taste, and freeze-dry to obtain the Ophiopogon japonicus extract.

[0177] 2.2 Preparation process of microsponge:

[0178] Take an appropriate amount of Ophiopogon japonicus extract and ethyl cellulose and dissolve them in dichloromethane as phase A, and use acetone: water (1:5) as solvent to prepare xanthan gum solution (0.1% xanthan gum + 0.1% Tween 80 solution). Take an appropriate amount of xanthan gum solution and disperse it in phase A. Use an ultrasonic cell homogenizer with a power of 100W, work for 3s and then stand for 3s, and continuously homogenize until it is a uniform emulsion, which is the internal phase B. Take an appropriate amount of PVA in a beaker, add water to swell completely, as the external phase. Slowly add the internal phase B dropwise into the external phase, and use a magnetic stirrer to continue stirring until it is evenly dispersed. Place the resulting mixture in a fume hood to evaporate the dichloromethane, filter it, wash it twice with distilled water, and place it in a 60°C oven to dry to obtain the Ophiopogon japonicus extract microsponge.

[0179] 3. Comparative experiment: Set up five experimental groups, as follows:

[0180] Group 1: 300 μmol / mL corn silk extract

[0181] Group 2: 300 μmol / mL Ophiopogon japonicus extract

[0182] Group 3: Ophiopogon japonicus extract microsponges: Corn silk extract (1:1)

[0183] Group 4: Ophiopogon japonicus extract microsponge: Corn silk extract (1:2)

[0184] Group 5: Ophiopogon japonicus extract microsponges: Corn silk extract (2:1)

[0185] 3.1 Cell experiments in each experimental group

[0186] HaCaT cells were cultured in HaCaT medium at 37°C and 5% CO 2 The cells were cultured in an incubator at 4 °C. 0.25% trypsin solution was used for cell digestion and passage, and the cells were passaged every 2 to 3 days. The cells were incubated with 80 μg / mL sodium dodecyl sulfate (SLS) for 4 h as a skin barrier damage model.

[0187] The cells were incubated with different ratios of Ophiopogon japonicus extract and Zea mays extract for 24 h.

[0188] The grouped HaCaT cells were inoculated into 96-well culture plates and incubated for 24 hours according to the experimental design. Then 10 μL of CCK-8 working solution was added, and the cells were cultured for another 2 hours at 37°C, and then the absorbance of the cells at a wavelength of 450 nm was measured on an ELISA reader. The proliferation rate was calculated based on the absorbance value.

[0189] Fig. 9 The cell proliferation rate results are shown in Figure 2. Fig. 9 It can be seen that after SLS treatment, HaCaT cells undergo abnormal proliferation, and after drug intervention, proliferation is inhibited to varying degrees, and group 3 (the composition of the present invention) has the best inhibitory effect.

[0190] 4. The animal experiment of the composition of the present invention was conducted by setting up six experimental groups, namely, a low-dose group, a medium-dose group, a high-dose group, a matrix group, a blank group, and a model group. The specific method is as follows:

[0191] 40 ICR mice were divided into groups according to the following examples, with 8 mice in each group. The 3*3cm area on the back of each group of mice was depilated (after shaving, the depilatory cream was used to depilate again after 1 day). The skin barrier damage model was established in each group of mice using the tape method, twice a day (8:30 and 16:00), and the operation was repeated 5 times each time until the skin became dry and scaly, and red spots were congested. After the operation, the experimental drugs were given to each group except the blank model group, twice a day, 0.5g / mouse, for 7 days. After 7 days, the mice were killed and the back skin was taken.

[0192] Low-dose group: the sample (microsponge of Ophiopogon japonicus extract: fermented corn silk 1:1) with Ophiopogon japonicus concentration of 4 mg / g and fermented corn silk 4 mg / g were dissolved in 5 mL of water, and 35 g of 1% carbomer, 5 g of propylene glycol and 5 g of glycerol were added to prepare 50 g of medicated gel.

[0193] Medium-dose group: Dissolve the sample (microsponge of Ophiopogon japonicus extract: fermented corn silk in a ratio of 1:1) with Ophiopogon japonicus at a concentration of 20 mg / g and fermented corn silk at 20 mg / g in 5 mL of water, add 35 g of 1% carbomer, 5 g of propylene glycol and 5 g of glycerol to prepare 50 g of medicated gel.

[0194] High-dose group: the sample (microsponge of Ophiopogon japonicus extract: fermented corn silk 1:1) with Ophiopogon japonicus concentration of 40 mg / g and fermented corn silk 40 mg / g were dissolved in 5 mL of water, and 35 g of 1% carbomer, 5 g of propylene glycol and 5 g of glycerol were added to prepare 50 g of medicated gel.

[0195] Matrix group: 35 g of 1% carbomer, 5 g of propylene glycol, and 5 g of glycerol were added to 5 mL of water to prepare 50 g of drug-containing gel.

[0196] Blank group: saline group

[0197] After modeling, no medication was given to the model group.

[0198] 4.1 HE staining of dorsal skin, determination of ceramide content, and determination of IL-6 inflammatory factor content

[0199] The above groups were evaluated by HE staining of the back skin, determination of ceramide content, and determination of IL-6 inflammatory factor content. Figures 10 to 15 As shown, Fig.10 HE staining results of the blank group. Fig.10 It can be seen that the skin tissue structure is basically normal, the epidermis structure is intact, the spinous layer thickness is uniform, and no abnormalities are found; the collagen fibers in the dermis are arranged neatly and tightly, and the skin appendages are abundant. The blue arrows indicate the hair follicles, and the yellow arrows indicate the sebaceous glands; no obvious inflammatory cell infiltration is found in the tissue; Fig.11 HE staining microscopy results of the model group. Fig.11 It can be seen that the skin tissue structure is moderately abnormal, the epidermal structure is missing in the field of view, scabs are visible, and there is a lot of cell necrosis, as shown by the black arrow; the collagen fibers in the dermis are arranged neatly and tightly, skin appendages are visible, and the blue arrow indicates the hair follicles; a small amount of inflammatory cell infiltration can be seen in the tissue, as shown by the red arrow. Fig.12 HE staining results of the solvent group. Fig.12It can be seen that the skin tissue structure is moderately abnormal, with crusts visible in the field of view, accompanied by a large amount of cell necrosis, as shown by the black arrow. The thickness of the stratum spinosum is uniform, and no abnormalities are found. The collagen fibers in the dermis are arranged neatly and tightly, and bleeding is visible locally, as shown by the orange arrow. The skin appendages are abundant, and the blue arrow indicates the hair follicles. A small amount of inflammatory cell infiltration can be seen in the tissue, as shown by the red arrow. Fig.13 The results of HE staining in the low-dose group are shown in Figure 2. Fig.13 It can be seen that the skin tissue structure is moderately abnormal, with local crusts visible in the field of vision, accompanied by partial cell necrosis, as shown by the black arrow. The thickness of the spinous layer is uniform, and no obvious abnormalities are found. The collagen fibers in the dermis are loosely arranged locally, as shown by the green arrow. Skin appendages are visible, and the blue arrow indicates the hair follicles. A small amount of inflammatory cell infiltration can be seen in the tissue, as shown by the red arrow. Fig.14 HE staining microscopy results of the medium-dose group. Fig.14 It can be seen that the skin tissue structure is slightly abnormal. The structure of the epidermis is clear in the field of vision, and the spinous layer is thinned, as shown by the black arrow. The collagen fibers in the dermis are loosely arranged locally, as shown by the green arrow. The skin appendages are abundant, with the blue arrow indicating the hair follicles and the yellow arrow indicating the sebaceous glands. No obvious inflammatory cell infiltration was found in the tissue. Fig.15 HE staining microscopy results of the high-dose group. Fig.15 It can be seen that the skin tissue structure is moderately abnormal. The structure of the epidermis is clear in the field of vision, and the spinous layer is thinned, as shown by the black arrow; the collagen fibers in the dermis are loosely arranged and the content is reduced, as shown by the green arrow. The skin appendages are abundant, with the blue arrow indicating the hair follicles and the yellow arrow indicating the sebaceous glands; no obvious inflammatory cell infiltration is seen in the tissue.

[0200] from Figures 10 to 15 It can be seen that after HE staining microscopy, the skin tissue structure of the model group was moderately abnormal, the epidermal structure was missing in the visual field, and a small amount of inflammatory cell infiltration was seen in the tissue, confirming that the skin was damaged and the skin barrier was damaged. The model was successfully established. After administration of the high, medium and low dose groups, the skin tissue structure of the medium dose group was slightly abnormal, the epidermal structure was clear in the visual field, and no obvious inflammatory cell infiltration was seen in the tissue, confirming that it had a skin repair effect. This shows that the composition has a repairing effect on damaged skin barrier.

[0201] 4.2. Results of ELISA determination of ceramide content in dorsal skin tissue

[0202] As a key component of intercellular lipids in the stratum corneum, ceramide is essential for maintaining the skin barrier function. It mainly strengthens the connection between keratinocytes to form a waterproof barrier, effectively prevents water loss, and improves the skin's water retention capacity. At the same time, ceramide can promote cell growth, increase the thickness of the stratum corneum, and enhance the skin's resistance to external stimuli. Therefore, the level of ceramide in the skin directly reflects the level of the skin barrier.

[0203] Fig.16 It is a graph showing the measurement results of the Cer content. From Fig.16 it can be seen that by measuring the ceramide content in the back skin tissues of mice in each group using the ELISA method, compared with the blank group, the ceramide content in the back skin tissues of the model group decreased significantly. After intervention, the ceramide content increased to varying degrees, and the ceramide content in the medium-dose group was the highest.

[0204] 4.3 Results of measuring the TNF-α content in the back skin tissues by the ELISA method

[0205] Fig.17 It is a graph showing the measurement results of the TNF-α content. It can be seen that the content of the inflammatory factor TNF-α in the back skin tissues of mice. Compared with the blank group, the TNF-α content in the back skin tissues of the model group increased significantly. After intervention, the TNF-α content decreased to varying degrees, and the decrease degree in the medium-dose group was the most significant.

[0206] 4.4 Skin external preparation prepared from the composition of the present invention

[0207] In this example, the corn silk extract and the ophiopogon japonicus extract microsponge in a ratio of 1:1 are used as the core components for the preparation of a skin external preparation. The skin external preparation is preferably a cosmetic composition, such as a face cream, essence, milk cream, etc. The preferred weight percentage of the composition is 0.01%-10% (w / w). The specific preparation process of the face cream is shown in Table 2, the preparation process of the emulsion is shown in Table 3, and the preparation process of the essence is shown in Table 4.

[0208] Table 2 Specific preparation process of the face cream

[0209]

[0210]

[0211] Table 3 Preparation process of the emulsion

[0212]

[0213] Table 4 Preparation process of the essence

[0214]

[0215]

Claims

1. A skin dressing composition, characterized in that The skin external dressing composition is composed of corn silk extract and ophiopogon japonicus extract micro-sponges in a weight ratio of 1:

1.

2. The skin external dressing composition according to claim 1, characterized in that The preparation method of the corn silk extract is:

1. The crushed dry corn silk is mixed with water to form a suspension, sterilized twice, and then inoculated with a mixed bacteria of Bifidobacterium adolescentis and yeast for fermentation to obtain a fermentation product; 2. Add 70% ethanol to the fermented product for ultrasonic extraction, then filter, collect the filtrate, evaporate and concentrate, and vacuum freeze-dry to obtain corn silk extract.

3. The skin external dressing composition according to claim 2, characterized in that The material-liquid ratio of the crushed dry corn silk to water in step 1 is 60g:40L.

4. The skin external dressing composition according to claim 2, characterized in that In step 1, the inoculation amount is a mixture of 5% Bifidobacterium adolescentis fermentation broth and 5% yeast fermentation broth, and the ratio of the number of live bacteria in the Bifidobacterium adolescentis fermentation broth to that in the yeast fermentation broth is 1:

1.

5. The skin external dressing composition according to claim 2, characterized in that In step 2, the material-liquid ratio of the fermentation product to 70% ethanol is 1:12 g / mL, the ultrasonic extraction temperature is 50° C., and the ultrasonic time is 30 min.

6. The skin external dressing composition according to claim 1, characterized in that The preparation method of the Ophiopogon japonicus extract micro-sponge:

1. Preparation of Ophiopogon japonicus extract: add 80% ethanol to reflux extract of Ophiopogon japonicus powder, combine the extracts and filter, evaporate to remove ethanol until there is no alcohol taste, and freeze-dry to obtain Ophiopogon japonicus extract; 2. Dissolve Ophiopogon japonicus extract and ethyl cellulose in dichloromethane to obtain phase A, mix acetone and water to prepare xanthan gum Tween 80 solution; disperse the xanthan gum Tween 80 solution in phase A, and then homogenize with an ultrasonic cell homogenizer until it becomes a uniform emulsion to obtain inner phase B; add water to PVA to swell completely as the outer phase, add inner phase B dropwise into the outer phase and stir until it is evenly dispersed, then evaporate the dichloromethane, filter, wash and dry to obtain the Ophiopogon japonicus extract micro sponge.

7. The skin external dressing composition according to claim 6, characterized in that In step 1, 8 times 80% ethanol was added to the Ophiopogon japonicus powder and refluxed for extraction twice, each extraction was for 4 hours.

8. The skin external dressing composition according to claim 6, characterized in that In step 2, the volume ratio of acetone to water is 1:5, the concentration of xanthan gum in the xanthan gum Tween 80 solution is 0.1%, and the concentration of the Tween 80 solution is 0.1%.

9. The skin external dressing composition according to claim 6, characterized in that In step 2, the xanthan gum Tween 80 solution is dispersed in phase A, and the ultrasonic cell homogenizer is used at a power of 100 W, working for 3 seconds and then resting for 3 seconds, and continuously homogenizing until it becomes a uniform emulsion.

10. The skin external dressing composition according to claim 6, characterized in that In step 2, the dichloromethane is evaporated, filtered, washed and dried, that is, after the dichloromethane is evaporated in a fume hood, it is filtered, washed twice with distilled water, and dried in an oven at 60°C.

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