Composition containing typha orientalis as well as preparation method and application of composition

Abstract

The invention relates to the technical field of cosmetics, in particular to a composition containing typha orientalis as well as a preparation method and application of the composition. The preparation method comprises the following steps: fermenting pollen typhae extract freeze-dried powder by using lactobacillus plantarum and preparing a typha orientalis extract lactobacillus plantarum fermented product; fermenting scutellaria baicalensis root extract freeze-dried powder by using aspergillus niger and preparing a scutellaria baicalensis root extract aspergillus niger fermented product; mixing the typha orientalis extract lactobacillus plantarum fermentation product with the scutellaria baicalensis root extract aspergillus niger fermentation product to obtain a composition containing typha orientalis; on one hand, the lactobacillus plantarum is used for fermenting the typha orientalis extract, so that the typha orientalis extract can be better converted into a substance with higher activity; on the other hand, the aspergillus niger is used for fermenting the scutellaria baicalensis root extract, low-activity glycoside is converted into flavonoid aglycone with higher biological activity, and the variety and activity of fermentation products are increased; the combined fermentation process is carried out under mild conditions, so that the damage of high temperature, high pressure and the like to active ingredients is effectively avoided, and the biological activity of the typha orientalis and scutellaria baicalensis root extract is completely reserved.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a composition containing cattail, its preparation method, and its application. Background Technology

[0002] Typha pollen is the pollen of Typha orientalis, a plant belonging to the Typhaceae family. Typha angustifolia L .), Oriental cattail ( Typha orientalis Presl (or dried pollen of plants in the same genus). Typha pollen extract has antioxidant and anti-inflammatory effects. For example, Chinese patent 202410329627.5 discloses the application of *Typha orientalis* in the preparation of a composition for inhibiting skin pigmentation. It describes mixing dried *Typha orientalis* pollen with 20-40 times its weight of a 20-30 wt% butylene glycol aqueous solution, homogenizing the mixture in a microfluidic homogenizer for 10-20 minutes, and then filtering the homogenized material to obtain a crude liquid. The homogenization pressure of the microfluidic homogenizer in step A is 80-130 MPa, the homogenization temperature is 40-65℃, and the homogenization time is 5-100 min.

[0003] The shortcoming of existing technology is that the homogenization effect of high pressure and high temperature will significantly reduce the bioactive components, causing irreversible damage, and thus reducing the efficacy of the composition. Summary of the Invention

[0004] In view of the shortcomings of the prior art, one objective of the present invention is to provide a method for preparing a composition containing oriental cattail, another objective of the present invention is to provide a composition containing oriental cattail, and a third objective of the present invention is to provide the use of a composition containing oriental cattail in cosmetics.

[0005] One of the objectives of this invention is achieved through the following technical solution: A method for preparing a composition containing oriental cattail includes the following steps: Step 1, use Oriental cattail ( Typha orientalis Presl To prepare freeze-dried powder of Typha pollen extract from dried pollen of Scutellaria baicalensis, scutellaria baicalensis ( ) was used. Scutellaria baicalensis Georgi Scutellaria baicalensis root extract freeze-dried powder was prepared from dried roots. Step 2: Ferment the freeze-dried powder of Typha pollen extract with Lactobacillus plantarum to prepare Typha orientalis extract Lactobacillus plantarum ferment, and ferment the freeze-dried powder of Scutellaria baicalensis root extract with Aspergillus niger to prepare Scutellaria baicalensis root extract Aspergillus niger ferment. Step 3: Mix the fermented product of *Typha orientalis* extract and *Aspergillus niger* fermented product of *Scutellaria baicalensis* root extract to obtain a composition containing *Typha orientalis*.

[0006] Furthermore, in step one, the method for preparing the lyophilized powder of Typhae pollen extract includes: Weigh 200g of dried pollen from *Typha orientalis*, add 70% ethanol at a material-to-liquid ratio of 1:10, soak the herb for 1-2 hours, heat in a 75℃ water bath for 2-3 hours, filter and collect the filtrate, transfer the filtrate to a rotary evaporator, and concentrate under reduced pressure to 1 / 6-1 / 4 of the original liquid volume. Adjust the pH of the concentrate to 9-10 with 5% NaOH and store at 4℃ for 12 hours. The next day, take out the sample, allow it to return to room temperature, centrifuge at 8000 rpm for 10 minutes, collect the supernatant, adjust the pH to approximately 2 with 5% HCl, and store at 4℃ for 12 hours. The next day, take out the sample, allow it to return to room temperature, centrifuge at 10000 rpm for 10 minutes, retain the precipitate, and freeze-dry to obtain the lyophilized powder of *Typha orientalis* extract.

[0007] Furthermore, in step one, the method for preparing the lyophilized powder of Scutellaria baicalensis root extract includes: Weigh 200g of dried Scutellaria baicalensis root, pulverize it using a traditional Chinese medicine pulverizer, and collect the powder passing through a 40-mesh sieve. Soak the medicinal material in 70% ethanol at a material-to-liquid ratio of 1:20 for 1-2 hours, heat in a 75℃ water bath for 2-3 hours, filter, and collect the filtrate. Transfer the filtrate to a rotary evaporator and concentrate under reduced pressure to 1 / 6-1 / 4 of the original liquid volume. Adjust the pH of the concentrate to 9-10 with 5% NaOH and store at 4℃ for 12 hours. The next day, remove the sample, allow it to return to room temperature, centrifuge at 8000 rpm for 10 minutes, collect the supernatant, adjust the pH to approximately 2 with 5% HCl, and store at 4℃ for 12 hours. The next day, remove the sample, allow it to return to room temperature, centrifuge at 10000 rpm for 10 minutes, retain the precipitate, and freeze-dry to obtain the freeze-dried powder of Scutellaria baicalensis root extract.

[0008] Furthermore, in step two, the *Lactobacillus plantarum* is GDMCC 1.2868 (… Lactobacillus acidophilus The strain was purchased from the Guangdong Provincial Center for Microbial Culture Collection; the Aspergillus niger was GDMCC 1.2374, which was also purchased from the Guangdong Provincial Center for Microbial Culture Collection.

[0009] Further, step two includes activation of *Lactobacillus plantarum*: a small amount of *Lactobacillus plantarum* is taken from a glycerol tube and inoculated into MRS liquid medium, and cultured on a shaker at 37°C and 150 rpm for 24 hours for primary activation. The primary activated bacterial solution is then transferred to fresh MRS liquid medium at a 1% inoculation rate and cultured under the same conditions for 12 hours to complete secondary activation, ensuring that *Lactobacillus plantarum* is in a good growth activity state, and the activated *Lactobacillus plantarum* is obtained for later use; it also includes activation of *Aspergillus niger*: *Aspergillus niger* slant culture is transferred to PDA plate medium and cultured in a constant temperature incubator at 30°C for 48-72 hours. When white mycelia grow on the plate and black conidia are produced, single colonies with neat edges and vigorous growth are selected, transferred to PDA liquid medium, and cultured on a shaker at 30°C and 180 rpm for 24-36 hours to complete the activation and rejuvenation of *Aspergillus niger*, and it is then ready for use.

[0010] Furthermore, in step two, the method for fermenting and preparing *Lactobacillus plantarum* ferment of *Typha orientalis* extract includes: A liquid culture medium containing 0.4-1% (w / v) of lyophilized Typha orientalis extract was prepared using pure water. The inoculum of activated Lactobacillus plantarum was 1-16% (v / v). The fermentation system was placed in a shaker at 37℃ and a rotation speed of 150-180 r / min for 48 h. Fermentation was then terminated by filtration, and the filtrate was retained to obtain the Typha orientalis extract Lactobacillus plantarum fermentation product. The fermentation product was transferred to a rotary evaporator and concentrated under reduced pressure at 45-55℃ until the volume of the concentrated liquid was 1 / 10-1 / 5 of the original volume. The concentrated liquid was then freeze-dried for later use.

[0011] Furthermore, in step two, the method for fermenting and preparing the Aspergillus niger ferment of Scutellaria baicalensis root extract includes: A liquid culture medium containing 0.4-1% (w / v) of lyophilized Scutellaria baicalensis root extract was prepared using pure water. The inoculum of activated Aspergillus niger was 1-16% (v / v). The fermentation system was cultured in a shaker at 28-30℃ at a speed of 100-150 rpm for 72 hours. The filtrate was filtered to obtain the Aspergillus niger ferment broth from the Scutellaria baicalensis root extract. The ferment broth was transferred to a rotary evaporator and concentrated under reduced pressure at 45-55℃ until the volume of the concentrated liquid was 1 / 10-1 / 5 of the original volume. The concentrated broth was then freeze-dried for later use.

[0012] Furthermore, in step three, the volume ratio of *Lactobacillus plantarum* fermentation product from *Typha orientalis* extract to *Aspergillus niger* fermentation product from *Scutellaria baicalensis* root extract is 7:3-8.

[0013] Furthermore, step three also includes adding Acanthopanax senticosus root extract, wherein the volume ratio of fermented Typha orientalis extract, fermented Scutellaria baicalensis root extract and Acanthopanax senticosus root extract is 7:5:1-5.

[0014] Furthermore, the preparation method of the Acanthopanax senticosus root extract includes: Acanthopanax senticosus ( Acanthopanax senticosus The dried roots of Acanthopanax senticosus were dried in an oven at 50-55℃ until constant weight. 200g of the powder was weighed and pulverized using a traditional Chinese medicine pulverizer, and the powder was collected and passed through a 40-mesh sieve. The lyophilized powder of Acanthopanax senticosus root extract was prepared using an ultrasonic-assisted water extraction method. The Acanthopanax senticosus root powder was mixed with purified water at a ratio of 1:30 (g / mL), with an ultrasonic power of 450-500W, an extraction temperature of 45-55℃, and an extraction time of 1.4-1.6 hours. After extraction, the mixture was filtered through a 40-mesh filter cloth, and the filtrate and residue were collected separately. The filtrate was transferred to a rotary evaporator and concentrated under reduced pressure at 65℃ until the volume of the concentrated liquid was 1 / 6-1 / 4 of the original volume, yielding the concentrated Acanthopanax senticosus extract. This concentrated extract was then freeze-dried to obtain the lyophilized powder of Acanthopanax senticosus root extract.

[0015] The second objective of this invention is achieved through the following technical solution: The above-described preparation method yields a composition containing cattail.

[0016] The third objective of this invention is achieved through the following technical solution: Cosmetics are obtained by using the above-mentioned composition containing cattail and by a preparation method permitted by the cosmetics industry.

[0017] The beneficial effects of this invention are as follows: *Lactobacillus plantarum*'s glycoside hydrolytic enzymes can convert low-activity flavonoid glycosides into more bioavailable flavonoid aglycones or water-soluble intermediates, enhancing core functions such as antioxidation and anti-inflammation, and increasing the activity of *Typha orientalis* extract. *Aspergillus niger* can secrete glycoside hydrolytic enzymes such as β-glucuronidase, cellulase, and pectinase, which can hydrolyze specific glycosidic bonds of flavonoid glycosides such as baicalin, wogonin, and baicalin, converting low-activity glycosides into more bioactive flavonoid aglycones, thus enhancing the activity of *Scutellaria baicalensis* root extract. Compared with single-strain fermentation or traditional extraction methods, this process achieves effective utilization of *Typha orientalis* and *Scutellaria baicalensis* root extracts, maximizing the release and conversion of their active ingredients. On the one hand, *Lactobacillus plantarum* fermentation of *Typha orientalis* extract can better convert it into more active substances; on the other hand, *Aspergillus niger* fermentation of *Scutellaria baicalensis* root extract converts low-activity glycosides into more bioactive flavonoid aglycones, increasing the variety and activity of fermentation products. Meanwhile, the co-fermentation process is carried out under mild conditions, which effectively avoids the damage to active ingredients caused by high temperature and high pressure, and fully preserves the bioactivity of the extracts of Typha orientalis and Scutellaria baicalensis. Detailed Implementation

[0018] The following is a further explanation with reference to specific implementation methods: Strains and medicinal materials Aspergillus niger GDMCC1.2374 and Lactobacillus plantarum GDMCC1.2868 ( Lactiplantibacillusplantarum Purchased from Guangdong Provincial Microbial Culture Collection Center.

[0019] Activation of Lactobacillus plantarum: Lactobacillus plantarum GDMCC1.2868 ( Lactobacillus acidophilus The bacteria were purchased from the Guangdong Provincial Microbial Culture Collection Center. A small amount of *Lactobacillus plantarum* was taken from a glycerol tube and inoculated into MRS liquid medium. The medium was then cultured at 37°C and 150 rpm for 24 hours for primary activation. The activated bacterial culture was then transferred to fresh MRS liquid medium at a 1% inoculum size and cultured under the same conditions for 12 hours to complete secondary activation, ensuring that *Lactobacillus plantarum* was in a good growth and active state. The activated *Lactobacillus plantarum* was then obtained for later use. Activation of Aspergillus niger: Aspergillus niger GDMCC1.2374 was purchased from Guangdong Provincial Microbial Culture Collection Center. The Aspergillus niger slant culture was transferred to PDA agar plates and incubated at 30℃ for 60 hours. Once white mycelia and black conidia appeared on the plates, single colonies with neat edges and vigorous growth were selected and transferred to PDA liquid medium. The colonies were then incubated at 30℃ and 180 rpm for 30 hours to complete the activation and rejuvenation of Aspergillus niger, and were ready for use.

[0020] The method for preparing lyophilized Typha pollen extract powder includes: weighing 200g of dried pollen from Typha orientalis, soaking the medicinal material in 70% ethanol at a material-to-liquid ratio of 1:10 for 1.5h, heating in a 75℃ water bath for 2.5h, filtering and collecting the filtrate, transferring the filtrate to a rotary evaporator, and concentrating under reduced pressure to 1 / 5 of the original liquid volume. The concentrated solution is adjusted to pH 9.5 with 5% NaOH and stored at 4℃ for 12h. The next day, the sample is taken out, and after the sample has returned to room temperature, it is centrifuged at 8000 rpm for 10min. The supernatant is taken, and the pH is adjusted to approximately 2 with 5% HCl and stored at 4℃ for 12h. The next day, the sample is taken out, and after the sample has returned to room temperature, it is centrifuged at 10000 rpm for 10min, retaining the precipitate, and freeze-drying to obtain lyophilized Typha pollen extract powder.

[0021] The method for preparing lyophilized powder of Scutellaria baicalensis root extract includes: weighing 200g of dried Scutellaria baicalensis root, pulverizing it using a traditional Chinese medicine pulverizer, and collecting the powder that passes through a 40-mesh sieve. Soak the medicinal material in 70% ethanol at a material-to-liquid ratio of 1:20 for 1.5 hours, heating in a water bath at 75℃ for 2.5 hours, filtering and collecting the filtrate. Transfer the filtrate to a rotary evaporator and concentrate under reduced pressure to 1 / 5 of the original liquid volume. Adjust the pH of the concentrate to 9.5 with 5% NaOH and store at 4℃ for 12 hours. The next day, remove the sample, allow it to return to room temperature, centrifuge at 8000 rpm for 10 minutes, collect the supernatant, adjust the pH to 2 with 5% HCl, and store at 4℃ for 12 hours. The next day, remove the sample, allow it to return to room temperature, centrifuge at 10000 rpm for 10 minutes, retain the precipitate, and freeze-dry to obtain lyophilized powder of Scutellaria baicalensis root extract.

[0022] Example 1

[0023] The method for fermenting and preparing *Lactobacillus plantarum* ferment broth from *Typha orientalis* extract includes: preparing a liquid culture medium with a *Typha orientalis* extract lyophilized powder content of 0.7% (w / v) using pure water; inoculating with 7.5% (v / v) of activated *Lactobacillus plantarum*; culturing the fermentation system in a shaker at 37℃ and a rotation speed of 165 r / min for 48 h; filtration to terminate fermentation; retaining the filtrate to obtain the *Typha orientalis* extract *Lactobacillus plantarum* ferment broth; transferring the ferment broth to a rotary evaporator and concentrating under reduced pressure at 50℃ until the volume of the concentrated liquid is 1 / 7 of the original volume; and then freeze-drying for later use.

[0024] The method for fermenting and preparing Aspergillus niger ferment of Scutellaria baicalensis root extract includes: preparing a liquid culture medium with a content of 0.7% (w / v) of lyophilized Scutellaria baicalensis root extract in pure water; inoculating 7.5% (v / v) of activated Aspergillus niger; and culturing the fermentation system in a shaker at 29℃ and 125 r / min for 72 h. The filtrate is filtered and retained to obtain the Aspergillus niger ferment of Scutellaria baicalensis root extract. The fermentation product is transferred to a rotary evaporator and concentrated under reduced pressure at 50℃ until the volume of the concentrated liquid is 1 / 7 of the original volume. The concentrated liquid is then freeze-dried for later use.

[0025] The fermented extract of Typha orientalis and the fermented extract of Scutellaria baicalensis were mixed at a volume ratio of 7:5.

[0026] Example 2

[0027] The difference from Example 1 is that in the step of preparing the fermentation product of Typha orientalis extract, a liquid culture medium with a Typha orientalis extract content of 0.7% (w / v) was prepared with pure water, and the inoculation amount of activated Lactobacillus plantarum was 1% (v / v). That is, the amount of Typha orientalis extract added was normal, but the inoculation amount of Lactobacillus plantarum was insufficient.

[0028] Example 3

[0029] The difference from Example 1 is that in the step of preparing the fermentation product of Typha orientalis extract, a liquid culture medium with a Typha orientalis extract freeze-dried powder content of 0.7% (w / v) was prepared with pure water, and the inoculation amount of activated Lactobacillus plantarum was 16% (v / v). That is, the amount of Typha orientalis extract added is normal, but the inoculation amount of Lactobacillus plantarum is too high.

[0030] Example 4

[0031] The difference from Example 1 is that in the step of preparing the fermentation product of Scutellaria baicalensis root extract, a liquid culture medium with a content of 0.7% (w / v) of Scutellaria baicalensis root extract was prepared with pure water, and the amount of activated Aspergillus niger inoculated was 1% (v / v). That is, the amount of Scutellaria baicalensis root extract added was normal, but the amount of Aspergillus niger inoculated was insufficient.

[0032] Example 5

[0033] The difference from Example 1 is that in the step of preparing the fermentation product of Scutellaria baicalensis root extract, a liquid culture medium with a content of 0.7% (w / v) of Scutellaria baicalensis root extract was prepared with pure water, and the amount of activated Aspergillus niger inoculated was 16% (v / v). That is, the amount of Scutellaria baicalensis root extract added was normal, but the amount of Aspergillus niger inoculated was too high.

[0034] Example 6

[0035] The difference from Example 1 is that the two fermentation steps are combined, and the fermentation product of Oriental cattail extract and the fermentation product of Scutellaria baicalensis root extract are mixed at a volume ratio of 7:8, that is, the proportion of Scutellaria baicalensis root extract in the mixed fermentation product is too high.

[0036] Example 7

[0037] The difference from Example 1 is that the two fermentation steps are combined, and the fermentation product of Oriental cattail extract and the fermentation product of Scutellaria baicalensis root extract are mixed at a volume ratio of 7:3, that is, the proportion of Oriental cattail fermentation product in the mixed fermentation product is too high.

[0038] Example 8

[0039] Based on Example 1, compositions 1-5 were prepared. The fermentation mixture (the volume ratio of the fermented product of Typha orientalis extract to the fermented product of Scutellaria baicalensis root extract was 7:5) was obtained according to Example 1. The fermentation mixture was combined with Acanthopanax senticosus root extract in a certain proportion to obtain compositions 1-5.

[0040] Composition 1 is composed of, by volume ratio, fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:2.

[0041] Composition 2 is composed of the following by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:3.

[0042] Composition 3 is in the following volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:4.

[0043] Composition 4 is composed of the following by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:1.

[0044] Composition 5 is in the following volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:5.

[0045] Preparation of Acanthopanax senticosus root extract: Acanthopanax senticosus roots were dried in an oven at 52.5℃ to constant weight. 200g of the dried root was pulverized using a traditional Chinese medicine pulverizer, and the powder was collected and passed through a 40-mesh sieve. The Acanthopanax senticosus root extract was then freeze-dried using an ultrasonic-assisted water extraction method. The Acanthopanax senticosus root powder was mixed with purified water at a ratio of 1:30 (g / mL), with an ultrasonic power of 475W, an extraction temperature of 50℃, and an extraction time of 1.5 h. After extraction, the mixture was filtered through a 40-mesh filter cloth, and the filtrate and residue were collected separately. The filtrate was transferred to a rotary evaporator and concentrated under reduced pressure at 65℃ until the volume of the concentrated liquid was 1 / 5 of the original volume, yielding the Acanthopanax senticosus extract concentrate. The concentrate was then freeze-dried to obtain the freeze-dried Acanthopanax senticosus root extract powder.

[0046] Determination of fermentation product activity in Examples 1-7 Experiment 1. Effects of fermentation products on the viability of photoaged HaCaT cells The system was divided into a normal group, a model group, and groups 1-7 (Examples 1-7), with 6 replicates per group. HaCaT cells at a density of 25,000 per well were seeded into 96-well plates and cultured for 24 hours. When the cell confluence reached 60%-70%, the old culture medium was discarded. 100 μL of basal culture medium was added to each well of the normal and model groups, and 100 μL of basal culture medium containing 360 μg / mL of the culture medium from Examples 1-7 was added to the remaining groups. The plates were then cultured for another 6 hours.

[0047] A photoaging cell model was constructed using UVA irradiation. The culture medium was discarded, and the cells were washed twice with PBS. 100 μL of PBS was added to each well to cover the cells, while the normal control group was covered with aluminum foil. UVA irradiation was then performed at a dose of 12.5 J / cm². After irradiation, the PBS was removed. 100 μL of basal DMEM culture medium was added to each well of the normal and model groups, and 100 μL of basal medium containing 360 μg / mL (as used in Examples 1-7) was added to each well of the remaining groups. Cells were cultured for another 24 h.

[0048] The effect of fermentation products from Examples 1-7 on the viability of photoaged HaCaT cells was detected using the MTT assay. After aspirating the culture medium, 100 μL of diluted 0.5 mg / mL MTT solution was added, and the cells were incubated at 37°C for 3 h. The MTT solution was then removed, and DMSO was added and shaken to dissolve the cells. The absorbance at 490 nm was measured using a microplate reader, and the cell viability was calculated with the normal group as a reference (results are shown in Table 1).

[0049] Table 1. Effects of fermentation products on the viability of photoaged HaCaT cells. Group Cell survival rate % Example 1 86.25 Example 2 68.24 Example 3 78.32 Example 4 70.42 Example 5 75.72 Example 6 83.06 Example 7 84.47 normal group 100 Model group 61.34 Experiment 2. Effects of fermentation products on the content of MMPs in photoaged HaCaT cells The experimental group setup, cell culture and drug administration, UVA irradiation to construct a photoaging cell model, and other steps were the same as in Experiment 1.

[0050] Analysis of MMP content in photoaged HaCaT cells. Cell culture supernatants from each group were collected and centrifuged at 12000 g (4℃, 10 min) to remove cell debris, yielding clear samples. Human MMPs were detected using an enzyme-linked immunosorbent assay (ELISA) kit. Standard gradient wells, blank control wells, and sample wells were set up according to the kit instructions. Data correction was performed using the absorbance of the blank wells as a baseline, and a standard curve was constructed. The pretreated supernatant was added to the detection wells, following the incubation-washing-color development reaction procedure. A standard concentration-OD value regression equation was established. The sample OD value was substituted into the equation to calculate the original concentration, and concentration correction was performed using the sample dilution factor. Finally, the MMP content in photoaged HaCaT cells was calculated (results are shown in Table 2).

[0051] Table 2. Effects of fermentation products on the content of MMPs in photoaged HaCaT cells. Group MMPs (pg / ml) Example 1 1243.21 Example 2 1619.37 Example 3 1466.85 Example 4 1585.62 Example 5 1541.25 Example 6 1390.63 Example 7 1301.88 normal group 1513.88 Model group 1782.49 Experiment 3. Fermentation products increased type I collagen content in photoaged HaCaT cells. The experimental group setup, cell culture and drug administration, UVA irradiation to construct a photoaging cell model, and other steps were the same as in Experiment 1.

[0052] Analysis of type I collagen content in photoaged HaCaT cells. Cell culture supernatants from each group were collected and centrifuged at 12000 g (4℃, 10 min) to remove cell debris, yielding clear samples. Quantitative detection was performed using a human type I collagen protein-linked immunosorbent assay (ELISA) kit. Standard gradient wells, blank control wells, and sample wells were set up according to the kit instructions. Data correction was performed using the absorbance of the blank wells as a baseline, and a standard curve was constructed. The pretreated supernatant was added to the detection wells, following the incubation-washing-color development reaction procedure. A standard concentration-OD value regression equation was established, and the original concentration was calculated by substituting the sample OD value into the equation. Concentration correction was performed using the sample dilution factor to finally obtain the MMP-1 content in photoaged HaCaT cells (results are shown in Table 3).

[0053] Table 3. Fermentation products increase type I collagen content in photoaged HaCaT cells. Group Type I collagen (pg / ml) Example 1 13929.44 Example 2 9328.22 Example 3 10923.31 Example 4 9726.93 Example 5 10555.21 Example 6 11751.53 Example 7 12549.07 normal group 11475.46 Model group 8070.55 Experiment 4. Fermentation product reduces MDA content in photoaged HaCaT cells. The experimental group setup, cell culture and drug administration, UVA irradiation to construct a photoaging cell model, and other steps were the same as in Experiment 1.

[0054] Analysis of MDA content in photoaged HaCaT cells. Cell culture supernatants from each group were collected and centrifuged at 12000 g (4℃, 10 min) to remove cell debris, yielding clear samples. Quantitative detection was performed using a human MDA enzyme-linked immunosorbent assay (ELISA) kit. Standard gradient wells, blank control wells, and sample wells were set up according to the kit instructions. Data correction was performed using the absorbance of the blank wells as a baseline, and a standard curve was constructed. The pretreated supernatant was added to the detection wells, following the incubation-washing-color development reaction procedure. A standard concentration-OD value regression equation was established. The original concentration was calculated by substituting the sample OD value into the equation, and concentration correction was performed using the sample dilution factor. The final MDA content of photoaged HaCaT cells was obtained (results are shown in Table 4).

[0055] Table 4. Fermentation products reduce MDA content in photoaged HaCaT cells. Group MDA (pmol / mg prot) Example 1 1713.82 Example 2 2745.74 Example 3 2373.41 Example 4 2611.72 Example 5 2298.93 Example 6 2128.72 Example 7 1915.95 normal group 2518.06 Model group 3320.21 Experiment 5. Fermentation product enhances SOD enzyme activity in photoaged HaCaT cells. Set up a normal group, a model group, and 1-7 example groups, with 6 duplicate holes in each group.

[0056] Cell culture and drug administration. HaCaT cells were seeded in 6-well plates at a density of 300,000 per well and cultured for 24 h. When the cell confluence reached 60%-70%, the culture medium was aspirated. 2 mL of basal culture medium was added to the normal and model groups, and 2 mL of basal culture medium containing 360 μg / mL of the culture medium from Examples 1-7 was added to each well of the remaining groups. The plates were then incubated for another 6 h.

[0057] A photoaging cell model was constructed using UVA irradiation. The culture medium was discarded, and the cells were washed twice with PBS. 1 mL of PBS was added to each well to cover the cells, and UVA irradiation was applied. The normal control group was covered with aluminum foil. After irradiation, the PBS was removed. 2 mL of basal culture medium was added to the normal control and model groups, and 2 mL of basal culture medium containing 360 μg / mL (as in Examples 1-7) was added to each well of the remaining groups. The cells were cultured for another 24 h.

[0058] The SOD enzyme activity of photoaged HaCaT cells was detected. After culture was terminated, the culture medium was discarded and the cells were washed twice with PBS. The cell suspensions from each group were collected into centrifuge tubes and adjusted according to cell density (…). Cells were lysed by sonication in an ice bath with pre-chilled lysis buffer at a volume ratio of 500:1, then centrifuged at 12000g for 15 min at 4°C. The supernatant was collected and stored on ice. The BCA protein assay kit was used to detect the OD value at 562nm using a microplate reader, and the protein concentration of the sample was calculated. The superoxide dismutase (SOD) assay kit was performed according to the instructions, and the OD value was detected at 560nm using a microplate reader to calculate the SOD enzyme activity (results are shown in Table 5).

[0059] Table 5. Fermentation products enhance SOD enzyme activity in photoaged HaCaT cells. Group SOD enzyme activity U / mg prot Example 1 55.85 Example 2 36.96 Example 3 47.74 Example 4 39.63 Example 5 45.18 Example 6 49.52 Example 7 51.63 normal group 34.52 Model group 25.74 Experiment 6. Detection of antioxidant activity of fermentation products The antioxidant activity of the fermentation products in Examples 1-7 was detected by the DPPH free radical scavenging experiment. The specific steps are as follows.

[0060] A blank control group, a control group, and groups 1-7 (360 μg / mL) were set up, with 6 replicates per group. Experiments were conducted in 96-well plates. 100 μl of methanol was added to each well of the blank control group, 100 μl of methanol was added to each well of the control group, and 100 μl of fermentation material at a concentration of 360 μg / mL was added to each well of groups 1-7. Except for the blank control group, 100 μl of 0.2 mM DPPH working solution was added to each well. The plates were incubated in the dark for 30 min, and the OD value was measured at 517 nm using a microplate reader. The DPPH free radical scavenging rate was calculated using the following formula (see Table 6): DPPH free radical scavenging rate % = (1 - (A sample - A blank) / A control) * 100%.

[0061] Table 6. Detection of antioxidant activity of fermentation products Group DPPH removal rate % Example 1 88.63 Example 2 60.52 Example 3 68.15 Example 4 64.24 Example 5 72.38 Example 6 78.63 Example 7 81.95 Control group 7.49 Experiment 7. Effects of Compositions 1-5 on Fructosamine Content in the BSA-Fructose Model Composition 1 is composed of, by volume ratio, fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:2.

[0062] Composition 1 is composed of the following by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:3.

[0063] Composition 1 is composed of the following by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:4.

[0064] Composition 1 is composed of the following by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:1.

[0065] Composition 1 is composed of the following components by volume ratio: fermented extract of Typha orientalis: fermented extract of Scutellaria baicalensis: extract of Acanthopanax senticosus root = 7:5:5.

[0066] Cell grouping and drug administration. Control group, MGO group, and combination groups 1-5 (360 μg / mL) were established, with 6 replicates per group. HaCaT cells at a density of 25,000 per well were seeded in 96-well plates and cultured for 24 h. When cell confluence reached 60%-70%, drug administration was performed: 70 μL of basal medium was added to each well of the Control and MGO groups, and 70 μL of basal medium containing the composition was added to each well of the combination groups 1-5. Culture was continued for another 6 h.

[0067] A non-enzymatic glycosylated HaCaT cell model was established using MGO. After culture, 30 μL of DMEM medium was added to the Control group, and 30 μL of DMEM medium containing 4 mM MGO was added to each well of the MGO group and combinations 1-5, and the cells were cultured for another 24 h.

[0068] The effect of compositions 1-5 on the viability of non-enzymatically glycosylated HaCaT cells was detected using the MTT assay. After aspirating the culture medium, 100 μL of diluted 0.5 mg / mL MTT solution was added, and the cells were incubated at 37°C for 3 h. After removing the MTT solution, DMSO was added and the cells were shaken to dissolve. The absorbance at 490 nm was measured using a microplate reader, and the cell viability was calculated with the normal group as a reference (results are shown in Table 7).

[0069] Table 7 Effect of the composition on fructosamine content in the BSA-fructose model Group Non-enzymatic glycosylated cell survival rate % Composition 1 73.53 Composition 2 76.28 Composition 3 83.84 Composition 4 85.37 Composition 5 72.95 Control group 100 MGO Group 70.43 Experiment 8. Effect of Compositions 1-5 on the content of advanced glycosylation products (AGEs) Construction of the BSA-fructose model. BSA (final concentration 20 mg / mL) and fructose (0.5 M) were co-incubated in a constant temperature water bath at 50±0.5℃. Compositions 1-5 with a concentration of 360 μg / mL were pre-incubated with BSA solution for 10 min, and then fructose was added to start the reaction. After incubation, the mixture was removed and cooled for subsequent determination of products in the non-enzymatic glycosylation stage.

[0070] Determination of AGEs (Advanced Glycation End Products) Content. A certain amount of the above-mentioned incubation solution was taken, and the fluorescence emission spectrum of the incubation solution was measured using a multi-functional microplate reader with an excitation wavelength of 350 nm and an emission wavelength of 450 nm. The average slit width for excitation and emission was 5 nm. The fluorescence intensity of the glycosylated protein samples was recorded, and the content of AGEs in the BSA-fructose model was calculated (see Table 8).

[0071] Table 8 Effect of the composition on the content of advanced glycation end products (AGEs) Group AGEs content Composition 1 59.57 Composition 2 53.29 Composition 3 49.53 Composition 4 47.61 Composition 5 57.67 Blank group 18.84 Model group 62.22 The embodiments and descriptions above are merely illustrative of the principles and preferred embodiments of the present invention. Various changes and modifications may be made to the present invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed.

Claims

1. A method of preparing a composition comprising Typha orientalis, characterized in that, The method comprises the following steps: Step 1, preparing a cattail pollen extract freeze-dried powder by using dry pollen of the cattail, and preparing a scirpusin extract freeze-dried powder by using dry roots of the scirpus; Step 2, fermenting the cattail pollen extract freeze-dried powder by using lactobacillus plantarum to prepare a cattail extract lactobacillus plantarum fermentate, and fermenting the scirpusin extract freeze-dried powder by using aspergillus niger to prepare a scirpusin extract aspergillus niger fermentate; Step 3, mixing the cattail extract lactobacillus plantarum fermentate and the scirpusin extract aspergillus niger fermentate to obtain a cattail composition.

2. The method of preparing a Typha orientalis-containing composition according to claim 1, characterized in that: In the step 1, the method for preparing the cattail pollen extract freeze-dried powder comprises the following steps: 200g of dry pollen of the cattail is weighed, and the medicinal material is soaked in 70% ethanol according to a material-liquid ratio of 1:10 for 1-2h, and heated in a water bath at 75℃ for 2-3h; the filtrate is collected by filtration, and then transferred to a rotary evaporator for concentration under reduced pressure to 1 / 6-1 / 4 of the original liquid volume; the concentrated solution is adjusted to a pH of 9-10 by 5% NaOH, and stored at 4℃ for 12h; the next day, the sample is taken out, and centrifuged at 8000r for 10min after the sample is returned to room temperature; the supernatant is adjusted to a pH of about 2 by 5% HCl, and stored at 4℃ for 12h; the next day, the sample is taken out, and centrifuged at 10000r for 10min after the sample is returned to room temperature; the precipitate is reserved, and then freeze-dried to obtain the cattail pollen extract freeze-dried powder.

3. The method of claim 1, wherein the Typha orientalis-containing composition is prepared by the steps of: In the step 1, the method for preparing the scirpusin extract freeze-dried powder comprises the following steps: 200g of dry roots of the scirpus is weighed, and the medicinal material is crushed by a traditional Chinese medicine crusher, and the powder passing through a 40-mesh sieve is collected; the medicinal material is soaked in 70% ethanol according to a material-liquid ratio of 1:20 for 1-2h, and heated in a water bath at 75℃ for 2-3h; the filtrate is collected by filtration, and then transferred to a rotary evaporator for concentration under reduced pressure to 1 / 6-1 / 4 of the original liquid volume; the concentrated solution is adjusted to a pH of 9-10 by 5% NaOH, and stored at 4℃ for 12h; the next day, the sample is taken out, and centrifuged at 8000r for 10min after the sample is returned to room temperature; the supernatant is adjusted to a pH of about 2 by 5% HCl, and stored at 4℃ for 12h; the next day, the sample is taken out, and centrifuged at 10000r for 10min after the sample is returned to room temperature; the precipitate is reserved, and then freeze-dried to obtain the scirpusin extract freeze-dried powder.

4. The method of claim 1, wherein the Typha orientalis-containing composition is prepared by the steps of: In the step 2, the lactobacillus plantarum is GDMCC 1.2868, and the aspergillus niger is GDMCC 1.2374.

5. The method of claim 1, wherein the Typha orientalis-containing composition is prepared by the steps of: The step two further comprises Lactobacillus plantarum activation: a small amount of Lactobacillus plantarum is taken from the glycerol tube and inoculated into MRS liquid medium, and is cultured at 37℃ and 150r / min for 24 hours to perform primary activation; the bacteria liquid after primary activation is inoculated into fresh MRS liquid medium at an inoculation amount of 1%, and is cultured under the same condition for 12 hours to complete secondary activation, so as to ensure that the Lactobacillus plantarum is in a good growth activity state, and activated Lactobacillus plantarum is obtained; the step two further comprises Aspergillus niger activation: Aspergillus niger slant strain is inoculated into PDA plate culture medium, and is cultured in a 30℃ constant temperature incubator for 48-72 hours; when white mycelium and black conidia are generated on the plate, a single colony with neat edge and vigorous growth is selected and inoculated into PDA liquid culture medium, and is cultured at 30℃ and 180r / min for 24-36 hours to complete the activation and rejuvenation of Aspergillus niger, and the activated Aspergillus niger is obtained.

6. The preparation method of the typha orientalis composition of claim 1, wherein: In the step two, the method for fermenting and preparing the typha orientalis extract Lactobacillus plantarum fermentate comprises: The typha orientalis extract freeze-dried powder is configured into liquid culture medium with a content of 0.4-1% (w / v) by using pure water, the inoculation amount of the activated Lactobacillus plantarum is 1-16% (v / v), the fermentation system is placed in a 37℃ shaking bed for culture, the rotation speed is 150-180r / min, and the fermentation is ended after 48 hours of fermentation; the filter liquid is reserved by filtration to obtain the typha orientalis extract Lactobacillus plantarum fermentate; the fermentate is transferred to a rotary evaporator for vacuum concentration under the condition of 45-55℃ until the volume of the concentrated liquid is 1 / 10-1 / 5 of the original volume, and the freeze-drying is prepared for standby use; In the step two, the method for fermenting and preparing the scutellaria baicalensis georgi root extract Aspergillus niger fermentate comprises: The scutellaria baicalensis georgi root extract freeze-dried powder is configured into liquid culture medium with a content of 0.4-1% (w / v) by using pure water, the inoculation amount of the activated Aspergillus niger is 1-16% (v / v), the fermentation system is placed in a 28-30℃ shaking bed for culture, the rotation speed is 100-150r / min, and the fermentation is ended after 72 hours of fermentation; the filter liquid is reserved by filtration to obtain the scutellaria baicalensis georgi root extract Aspergillus niger fermentate; the fermentate is transferred to a rotary evaporator for vacuum concentration under the condition of 45-55℃ until the volume of the concentrated liquid is 1 / 10-1 / 5 of the original volume, and the freeze-drying is prepared for standby use.

7. The method of claim 1, wherein the Typha orientalis-containing composition is prepared by the steps of: In the step three, the volume ratio of the typha orientalis extract Lactobacillus plantarum fermentate to the scutellaria baicalensis georgi root extract Aspergillus niger fermentate is 7:3-8.

8. The method of claim 7, wherein the Typha orientalis-containing composition is prepared by the steps of: The step three further comprises adding acanthopanax senticosus root extract, wherein the volume ratio of the typha orientalis extract fermentate, the scutellaria baicalensis georgi root extract fermentate and the acanthopanax senticosus root extract is 7:5:1-5; The preparation method of the acanthopanax senticosus root extract comprises: The dried roots of acanthopanax senticosus are dried to constant weight in an oven at 50-55°C, 200 g is weighed and crushed by a traditional Chinese medicine crushing machine, and the powder passing through a 40-mesh sieve is collected; an ultrasonic-assisted water extraction method is used to prepare acanthopanax senticosus root extract freeze-dried powder; the acanthopanax senticosus root powder is mixed with purified water at a material-to-liquid ratio of 1:30 (g / mL), the ultrasonic power is 450-500 W, the extraction temperature is 45-55°C, the extraction time is 1.4-1.6 h, after the extraction is completed, the filtrate and the residue are filtered with 40-mesh filter cloth and stored respectively; the filtrate is transferred to a rotary evaporator and concentrated at 65°C under reduced pressure until the volume of the concentrated liquid is 1 / 6-1 / 4 of the original volume, to obtain acanthopanax senticosus extraction concentrate; and the acanthopanax senticosus root extract freeze-dried powder is obtained by freeze-drying.

9. A composition comprising typha orientalis obtained by the method according to any one of claims 1 to 8.

10. A cosmetic obtained by a method allowed in the cosmetic industry using the composition comprising typha orientalis according to claim 9.