A sophora japonica pollen-based composition for promoting collagen production, and a preparation method and application thereof

Abstract

The present application relates to a kind of based on sophora japonica pollen composition for promoting collagen production and its preparation method and application, belong to sophora extract technical field.The present application is first prepared and applied in the preparation of composition to obtain sophora japonica pollen extract, and the preparation method of the sophora japonica pollen extract includes: taking sophora japonica powder, after boiling with water cooling, adding carbon source and aspergillus oryzae CICC 2066 is fermented, sterilization, filtration, concentration, freeze-drying is obtained.The sophora japonica pollen extract can be applied in food, also can be applied in cosmetics, can effectively promote the generation of collagen, increase skin elasticity and the effect of firmness.

Description

TECHNICAL FIELD

[0001] The present application relates to a sophora japonica L. pollen-based composition for promoting collagen production and a preparation method and application thereof, and belongs to the technical field of sophora japonica L. extract. BACKGROUND

[0002] The main active components of sophora japonica L. are flavonoids (such as rutin and quercetin), phenolic acids (such as chlorogenic acid), and a small amount of triterpenoid saponins. These components have strong antioxidant activity: flavonoids (especially quercetin) can scavenge reactive oxygen species (ROS) in the skin or tissues, reducing the oxidative damage of free radicals to fibroblasts, which are the core cells of collagen synthesis, and the impairment of their function will directly lead to a decrease in collagen synthesis. The phenolic hydroxyl structure can chelate metal ions (such as Fe 2+ , Cu 2+ ), inhibit lipid peroxidation mediated by metal ions, avoid abnormal peptide chain breakage or cross-linking of collagen, and maintain its structural stability.

[0003] The active components in sophora japonica L. extract can enhance the synthesis function of fibroblasts by interfering with cell signaling: quercetin can activate the MAPK / ERK pathway and the TGF-β / Smad pathway: the former promotes fibroblast proliferation, increasing the “cell base” of synthesized collagen; the latter is a key regulatory pathway for collagen synthesis, which can up-regulate the gene expression of type I and III collagen (COL1A1, COL3A1) and accelerate peptide chain synthesis. Some triterpenoid saponins can increase the activity of proline hydroxylase: the maturation of collagen depends on the hydroxylation of proline (to form a stable triple helix structure), and the enhancement of this enzyme activity can promote the hydroxylation reaction and reduce the accumulation of immature collagen. The content of collagen in the human body depends not only on synthesis but also on degradation rate, and sophora japonica L. extract can protect the synthesized collagen by inhibiting degradation enzymes. Stimulation such as ultraviolet rays and inflammation can induce excessive expression of matrix metalloproteinases (MMPs, such as MMP-1 and MMP-3), which can specifically hydrolyze the peptide bonds of collagen. Flavonoid components in sophora japonica L. can down-regulate the gene transcription of MMPs, reduce their activity, and reduce the decomposition of collagen. Phenolic acid components (such as chlorogenic acid) can inhibit the activity of elastase, which mainly degrades elastin but indirectly damages collagen fiber network when excessively activated, and its activity can maintain the stability of collagen organization distribution after being inhibited.

[0004] The polysaccharides and amino acids in the sophora flower extract can serve as nutrient precursors to support the synthesis of collagen: the polysaccharide components can regulate the structure of the extracellular matrix (ECM) around the fibroblasts, providing a suitable spatial environment for the assembly of collagen fibers (formation of fiber bundles). Some free amino acids (such as glycine and proline) are basic building blocks of collagen and can directly participate in the synthesis of peptide chains as raw materials, thereby indirectly improving the synthesis efficiency.

[0005] Fermentation can enhance the activity and richness of the plant extract and enhance the functional strength of the extract. At present, there are relatively few studies on the application of fermentation in the extraction of sophora flowers to enhance the promotion of collagen synthesis, and therefore, it is necessary to provide related research to verify the effect of such technology. Meanwhile, it is also necessary to further improve the efficiency of the sophora flower extract compound in promoting collagen synthesis. SUMMARY

[0006] In view of the above technical problems, the present application provides a sophora flower pollen-based composition for promoting collagen synthesis, a preparation method thereof and applications thereof.

[0007] In one aspect, the present application provides a preparation method of sophora flower pollen extract, which comprises: taking sophora flower powder, boiling with water, cooling, adding a carbon source and aspergillus oryzae for fermentation, sterilizing, filtering, concentrating and freeze-drying.

[0008] Further, the sophora flower pollen extract can be applied in food and also in cosmetics.

[0009] Further, the sophora flower pollen extract can effectively promote the synthesis of collagen and increase the elasticity and firmness of the skin.

[0010] Further, in the preparation method of the sophora flower pollen extract, the carbon source includes substances such as sucrose, glucose and fructose that can provide energy for bacterial fermentation. The aspergillus oryzae can be selected from aspergillus oryzae with the number CICC 2066, which can effectively and significantly enhance the effect of the sophora flower extract in promoting the synthesis of type I and type III collagen. The amount of the carbon source can be 0.01-3% of the weight of the sophora flower powder, and the amount of the aspergillus oryzae used for inoculation and fermentation can be 10 6 -10 11 g bacteria / g sophora flower powder. The fermentation temperature and time can be selected as 25-40℃ and 3-5 days.

[0011] The present application also provides a Sophora japonica pollen-based composition for promoting collagen production, which comprises Sophora japonica pollen extract, polypeptide, Cynara scolymus leaf extract, Nephehium lappaceum leaf extract and excipient.

[0012] Further, the excipient is water and polyhydric alcohol.

[0013] Further, the polypeptide comprises at least one of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, tripeptide-1, acetyl hexapeptide-8, blue copper peptide, oligopeptide-1, oligopeptide-2, hexapeptide-3 and hexapeptide-9.

[0014] Further, the polyhydric alcohol comprises at least one of ethylene glycol, propylene glycol, glycerol, butylene glycol, hexylene glycol and pentylene glycol.

[0015] Further, the composition comprises Sophora japonica pollen extract 0.01-2%, polypeptide 0.001-1%, Cynara scolymus leaf extract 0.01-1.5%, Nephehium lappaceum leaf extract 0.01-1.5%, polyhydric alcohol 15-40% and water.

[0016] Further, the preparation method of the Cynara scolymus leaf extract comprises the following steps: crushing Cynara scolymus leaves, adding 50-70wt% ethanol for extraction, filtering, concentrating and freeze-drying.

[0017] Further, the preparation method of the Nephehium lappaceum leaf extract comprises the following steps: crushing Nephehium lappaceum leaves, adding 35-60wt% ethanol for extraction, filtering, concentrating and freeze-drying.

[0018] The Cynara scolymus leaf extract and the Nephehium lappaceum leaf extract can produce an advantage complementary phenomenon, and can synergistically promote the endogenous production of collagen, effectively increase the skin elasticity and give the skin a full and young state.

[0019] The present application also provides a preparation method of the above-mentioned composition, which comprises the following steps: heating polyhydric alcohol, adding polypeptide for stirring, then adding water for continuous stirring, adding Sophora japonica pollen extract, Cynara scolymus leaf extract and Nephehium lappaceum leaf extract for stirring, sterilizing and obtaining the composition.

[0020] The present application also provides the use of the above-mentioned composition in preparing skin tightening and elasticity increasing cosmetic products.

[0021] The present application has the following beneficial effects compared to the prior art:

[0022] The present application prepares Sophora japonica pollen extract and applies it in the preparation of a composition. The preparation method of the Sophora japonica pollen extract comprises: taking Sophora japonica powder, boiling with water and cooling, adding a carbon source and Aspergillus oryzae CICC 2066 for fermentation, sterilization, filtration, concentration, and freeze-drying. The Sophora japonica pollen extract can effectively promote the generation of collagen, increase skin elasticity and tightening effect.

[0023] The Sophora japonica pollen extract of the present application can be applied in food and also in cosmetics, and has good application prospects.

[0024] The present application uses Sophora japonica pollen extract to prepare a composition, which can effectively enhance the effect of the composition in promoting collagen generation. And the fermentation method for preparing Sophora japonica pollen extract can further improve the effect of the composition in promoting collagen generation compared to other extraction methods.

[0025] The present application uses Aspergillus oryzae for fermentation, which can effectively improve the effect of the composition compared to other bacteria for fermentation, and the use of a specific Aspergillus oryzae with the number CICC 2066 can further improve the composition in promoting the synthesis of type I and type III collagen.

[0026] The present application uses Sophora japonica pollen extract, polypeptide, Cynara scolymus leaf extract, Rubus suavissimus leaf extract, and excipients to prepare a composition, which can effectively promote the generation of collagen, increase skin elasticity and tightening effect.

[0027] The Cynara scolymus leaf extract prepared by using 50-70wt% ethanol extraction and the Rubus suavissimus leaf extract prepared by using 35-60wt% ethanol extraction can both effectively promote the endogenous generation of type I and type III collagen. When they are used together, the effect of promoting the generation of type I and type III collagen is better than using only one of them, and they can synergistically enhance the generation of collagen. When the ratio of the two is within the range of 1:0.5-1.5, the effect is the best, and the skin tightening effect is optimal.

[0028] The composition prepared by the present application can effectively promote the generation of collagen at a low concentration, and can enhance skin elasticity and tightness. DETAILED DESCRIPTION

[0029] In order for those skilled in the art to have a clearer and more intuitive understanding of the present application, the present application will be further described below in conjunction with specific embodiments. The experimental methods not specified in the following examples are selected and operated according to conventional methods and conditions, or according to the instructions of the commodity.

[0030] Example 1

[0031] Preparation method of Cynara scolymus leaf extract: Cynara scolymus leaves were crushed, and 60wt% ethanol was added at a solid-liquid ratio of 1:15, and then extracted at 55°C and 80rpm for 5 hours. The extract was filtered through a 0.22μm filter membrane, concentrated at 50°C under reduced pressure to a filtrate weight of 10.4%, and then freeze-dried at -50°C to a water content of 1.42wt%.

[0032] Preparation method of Nephehium lappaceum leaf extract: Nephehium lappaceum leaves were crushed, and 50wt% ethanol was added at a solid-liquid ratio of 1:18, and then extracted at 52°C and 90rpm for 4 hours. The extract was filtered through a 0.22μm filter membrane, concentrated at 50°C under reduced pressure to a filtrate weight of 11.2%, and then freeze-dried at -50°C to a water content of 1.35wt%.

[0033] The above individual extracts and combinations were tested for their efficiency in promoting the production of collagen I and III. The combinations of the samples are shown in Table 1. In Table 1, Q represents the weight ratio of Cynara scolymus leaf extract to Nephehium lappaceum leaf extract.

[0034] Table 1: Sample composition

[0035]

[0036] The specific test method is as follows: HSF cells in the logarithmic growth phase were trypsinized to obtain a suspension, which was inoculated into a 12-well plate at 2×10 5 cells per well, and 1mL of culture medium was added for incubation in a 5% CO2, 37°C incubator for 24 hours. The culture medium was then aspirated, and the cells were washed with PBS three times. The PBS was then aspirated, and the subsequent experiment was performed. For the blank group, 150μL of PBS was added to each well, and then 1mL of culture medium was added. For the experimental group, 150μL of PBS was added to each well, and then samples (samples 1-11, with a final concentration of 100ppm) were added, and then 1mL of culture medium was added. The plates were then incubated in a 5% CO2, 37°C incubator for 72 hours. Each group was repeated four times. After incubation, the culture medium was aspirated, and the cells were washed with PBS three times. Lysis solution (RIPA) was added to each well, and the plate was shaken and centrifuged at 8000rpm for 10 minutes. The supernatant was then collected, and the content of collagen I and III was tested according to the ELISA kit instructions. The test results are shown in Table 2.

[0037] Table 2: Collagen I and III content

[0038]

[0039] According to the content of Table 2, the increase in collagen content of each group compared to the blank group was calculated as the value Δ. The results are shown in Table 3.

[0040] Table 3: The △ value of collagen in each group compared with the blank group

[0041]

[0042] According to the test results of Tables 2-3, the leaf extract of Cynara scolymus and the leaf extract of Raphiolepis indica prepared by the extraction method of the present application can effectively promote the endogenous production of collagen I and III. According to the comparison of samples 1-2 and 3-11, it can be known that after the two are combined, the effect of promoting the production of collagen I and III is better than using only one of them, and the two can synergistically enhance the production of collagen. According to the data between the samples, it can be obtained that the effect is best when the ratio of the two is in the range of 1:0.5-1.5, and the skin tightening and fullness effect is optimal.

[0043] Example 2

[0044] The extract in Example 1 is used to prepare a composition and related tests are carried out.

[0045] 1. Composition 1

[0046] The raw material composition is as follows in terms of weight percentage: 1.5% Sophora japonica pollen extract, 0.08% polypeptide, 0.5% Cynara scolymus leaf extract, 0.6% Raphiolepis indica leaf extract, 30% polyol, and the balance water; the polypeptide is composed of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, and palmitoyl tetrapeptide-7 in a weight ratio of 1:1:2; the polyol is composed of ethylene glycol, glycerol, and 1,3-butanediol in a weight ratio of 1:1:4.

[0047] The preparation method of Sophora japonica pollen extract is as follows: Sophora japonica pollen powder is taken, 20 times the weight of water is added, and after boiling, it is cooled to room temperature, 0.18% sucrose and Aspergillus oryzae CICC 2066 (5×10 7 bacteria / g Sophora japonica pollen) are added, and fermentation is carried out at 35°C and 60 rpm for 4.5 days (sterile air is introduced, and the air flow is 0.68 VVM), then sterilized, filtered through a 0.22 μm pore size filter membrane, concentrated to 11.07% of the weight of the filtrate at 58°C, then freeze-dried at -52°C to contain 1.95wt% water.

[0048] The preparation method of the composition is as follows: the polyol is placed in a container, heated to 45°C, the polypeptide is added, stirred at 100 rpm for 10 minutes, then water is added and continues to be stirred at the same speed for 25 minutes, Sophora japonica pollen extract, Cynara scolymus leaf extract, and Raphiolepis indica leaf extract are added and continue to be stirred at the same speed for 15 minutes, sterilized, and the composition is obtained.

[0049] 2. Composition 2

[0050] Raw material composition, according to percentage by weight: 1.3% Sophora japonica pollen extract, 0.095% polypeptide, 0.6% Cynara scolymus leaf extract, 0.62% Rubus suavissimus leaf extract, 35% polyol and the balance water; the polypeptide is composed of tripeptide-1, acetyl hexapeptide-8 and blue copper peptide according to a weight ratio of 1:2:3; the polyol is composed of ethylene glycol and 1,3-butanediol according to a weight ratio of 1:3.

[0051] The preparation method of Sophora japonica pollen extract is as follows: Sophora japonica pollen powder is taken, boiled with 25 times the weight of water, cooled to room temperature, 0.2% sucrose and Aspergillus oryzae CICC 2066 (added amount is 8×10 7 bacteria / g Sophora japonica pollen) are added, and fermentation is carried out at 35°C and 60 rpm for 3.5 days (sterile air is introduced, and the air flow is 0.65VVM), then sterilized, filtered by a 0.22μm pore size filter membrane, concentrated to 11.35% of the weight of the filtrate at 58℃, then freeze-dried at-52℃ to obtain a product containing 1.82wt% of water.

[0052] The preparation method of composition 2 is the same as that of composition 1.

[0053] 3. Composition 3

[0054] The difference from composition 1 is that Sophora japonica pollen extract is not used: raw material composition, according to percentage by weight: 0.08% polypeptide, 0.5% Cynara scolymus leaf extract, 0.6% Rubus suavissimus leaf extract, 30% polyol and the balance water; the polypeptide is composed of palmitoyl tripeptide-1, palmitoyl pentapeptide-4 and palmitoyl tetrapeptide-7 according to a weight ratio of 1:1:2; the polyol is composed of ethylene glycol, glycerol and 1,3-butanediol according to a weight ratio of 1:1:4.

[0055] The preparation method of the composition is as follows: the polyol is placed in a container, heated to 45℃, the polypeptide is added, stirred at 100 rpm for 10 minutes, then water is added and stirring is continued at the same speed for 25 minutes, Cynara scolymus leaf extract and Rubus suavissimus leaf extract are added and stirring is continued at the same speed for 15 minutes, sterilized to obtain the composition.

[0056] 4. Composition 4

[0057] The difference from composition 1 is that Sophora japonica pollen extract is prepared by fermentation with other numbered Aspergillus oryzae, specifically, Aspergillus oryzae with the number GDMCC NO.3.31 is used to replace Aspergillus oryzae in composition 1, and the others are the same.

[0058] 5. Composition 5

[0059] The difference from composition 1 is that the Sophora japonica pollen extract is prepared by fermentation using other bacteria, specifically Bacillus subtilis with number GDMCC NO. 1.258 replacing Aspergillus oryzae in composition 1, and the others are the same.

[0060] 6, composition 6

[0061] The difference from composition 1 is that the Sophora japonica pollen extract is prepared by fermentation using other bacteria, specifically Acetobacter with number CICC 20441 replacing Aspergillus oryzae in composition 1, and the others are the same.

[0062] 7, composition 7

[0063] The difference from composition 1 is that the preparation method of Sophora japonica pollen extract is different, and the others are the same.

[0064] The preparation method of Sophora japonica pollen extract is as follows: take Sophora japonica pollen powder, add 15 times the mass of 25wt% ethanol with a concentration of 25wt%, stir at 45℃ with 80rpm for 4 hours, filter with a 0.22μm pore size filter membrane, concentrate to 11.35% of the filtrate weight at 54℃, and then freeze-dry at -52℃ to contain 1.89wt% of water.

[0065] The above compositions are tested for performance, and the efficiency of the compositions in promoting collagen production is tested.

[0066] Take HSF cells in the logarithmic growth phase, trypsinize, and obtain a suspension, inoculate 2×10 5 cells per well into a 12-well plate, add 1mL of culture medium to a 5% CO2, 37℃ incubator for 24 hours, aspirate the culture medium, wash 3 times with PBS, aspirate the PBS wash, and proceed to the subsequent experiment. Blank group: continue to add 150μL of PBS per well, then add culture medium to 1mL; experimental group: continue to add 150μL of PBS per well, then add composition (composition 1-7, final concentration of 0.45wt%) and add culture medium to 1mL; place the above well plates in a 5% CO2, 37℃ incubator for continuous culture for 72 hours; each group is repeated 4 times. After the culture is completed, aspirate the culture medium, wash 3 times with PBS, add lysis solution (RIPA to 1mL) and shake, then centrifuge at 8000rpm for 10 minutes, take the supernatant, and test the contents of collagen I and III according to the contents of the ELISA kit instructions. The test results are shown in Table 4.

[0067] Table 4: Effect of promoting collagen production

[0068]

[0069] According to the test result of comparison 4, the composition prepared by the application can effectively promote the generation of collagen at low concentration, and can enhance the elasticity and tightness of the skin.

[0070] According to the comparison of compositions 1, 3 and 7, the application can effectively enhance the effect of the composition on promoting the generation of collagen by using sophora japonica pollen extract to prepare the composition. Compared with other extraction methods, the fermentation method for preparing sophora japonica pollen extract can further improve the effect of the composition on promoting the generation of collagen.

[0071] According to the comparison of compositions 1, 4-6, the application can effectively improve the effect of the composition by using aspergillus oryzae for fermentation, and further improve the effect of the composition on promoting the synthesis of type I and type III collagen by using aspergillus oryzae with a specific number of CICC 2066.

[0072] The above description of the embodiments is for the purpose of facilitating the understanding and application of the application by the ordinary skilled person in the art. Those skilled in the art can obviously make various modifications to the embodiments, and apply the general principles described herein to other embodiments without creative labor. Therefore, the application is not limited to the embodiments herein, and the improvements and modifications made by those skilled in the art to the application according to simple modifications or substitutions of the application should be within the protection scope of the application.

Claims

1. A composition based on Sophora japonica pollen that promotes collagen production, characterized in that, The composition comprises, by weight percentage, 0.01-2% Sophora japonica pollen extract, 0.001-1% polypeptide, 0.01-1.5% artichoke leaf extract, 0.01-1.5% rambutan leaf extract, 15-40% polyol and the balance water. The preparation method of Sophora japonica flower powder extract includes: taking Sophora japonica flower powder, adding water and boiling it, cooling it, adding a carbon source and Aspergillus oryzae for fermentation, sterilizing, filtering, concentrating, and freeze-drying; the Aspergillus oryzae is Aspergillus oryzae CICC 2066; The weight ratio of artichoke leaf extract to rambutan leaf extract is 1:0.8-1.5; The preparation method of artichoke leaf extract includes: crushing artichoke leaves, adding 50-70wt% ethanol for extraction, filtering, concentrating, and freeze-drying. The preparation method of rambutan leaf extract includes: crushing rambutan leaves, adding 35-60wt% ethanol for extraction, filtering, concentrating, and freeze-drying.

2. The composition according to claim 1, characterized in that, The amount of carbon source used is 0.01-3% of the weight of Sophora japonica flower powder, and the fermentation temperature and time are 25-40℃ for 3-5 days.

3. The composition according to claim 1, characterized in that, The dosage of Aspergillus oryzae used for inoculation and fermentation is 10. 6 -10 11 One bacterium / g of Sophora japonica pollen.

4. The composition according to claim 1, characterized in that, The polypeptide includes at least one of palmitoyl tripeptide-1, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, tripeptide-1, acetyl hexapeptide-8, blue copper peptide, oligopeptide-1, oligopeptide-2, hexapeptide-3, and hexapeptide-9; And / or, the polyols include at least one of ethylene glycol, propylene glycol, glycerol, butylene glycol, hexanediol, and pentanediol.

5. A method for preparing the composition according to any one of claims 1-4, characterized in that, The process involves heating a polyol, adding a polypeptide and stirring, then adding water and continuing to stir, adding sophora japonica pollen extract, artichoke leaf extract, and rambutan leaf extract and stirring, followed by sterilization to obtain the composition.

6. The use of the composition according to any one of claims 1-4 in the preparation of a product that promotes collagen production.

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