An anti-aging composition, its preparation method and applications

Abstract

The present invention discloses an anti-aging composition, its preparation method and application, belonging to the technical field of cosmetics. The composition includes extracts of Clerodendrum inerme and Valeriana officinalis L. root. Through the compounding of the extracts of Clerodendrum inerme and Valeriana officinalis L. root, the present invention can have synergistic effects, improve cell viability, reduce the expression of cell senescence genes, and has a significant anti-aging effect. Moreover, the composition of the present invention is a pure plant extract, with good safety, simple preparation method and low cost, and is suitable for industrial promotion.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and particularly relates to an anti-aging composition, a preparation method thereof and an application thereof. Background Art

[0002] In recent years, components that can delay skin aging have been found in some natural green plants. These components generally have the characteristics of safety and high efficiency, and are widely used by researchers in cosmetics.

[0003] CN110731928A discloses an anti-aging cosmetic composition, which discloses that birch sap with a concentration multiple of 1.05 - 8 times has the effect of anti-aging. CN109394591A discloses an anti-aging cosmetic process, which uses ethanol as an extraction solvent and ultrasonic extraction to obtain a seaweed extract that can inhibit the activity of elastase and scavenge free radicals, and has the effect of delaying skin aging. CN111643427A discloses an anti-aging composition, comprising the following components: red ginseng extract, ginseng root water, turmeric root extract, cistanche extract, dendrobium officinale stem extract, ganoderma lucidum extract, poria cocos extract, cornel extract, epimedium extract and polygonum cuspidatum root extract. This anti-aging composition has good antioxidant properties and the effect of activating cells, can prevent skin aging, and can effectively improve the aging condition of the skin, and has excellent anti-aging effects.

[0004] Today, when advocating a green lifestyle, people have a certain degree of rejection of chemically synthesized cosmetic raw materials. Compared with artificially synthesized anti-aging compounds, natural plant extracts meet the consumption concept of most consumers for green and environmentally friendly cosmetics. Coupled with its good efficacy, it has developed rapidly in recent years, and such cosmetics will also occupy an important position in the daily chemical field market.

[0005] The mangrove plant Clerodendrum inerme belongs to the genus Clerodendrum of the Verbenaceae family. It is mainly distributed in the southern coasts of Zhejiang, Fujian, Guangdong, Hainan and Guangxi in China, and is a sand-fixing and afforestation tree species in the southern coast of China. Clerodendrum inerme has certain medicinal value. Its roots can be used as medicine, which can clear heat and detoxify, and relax tendons and activate collaterals. As a traditional Thai medicine, fresh Clerodendrum inerme leaves are used to treat skin diseases. The powder of Clerodendrum inerme leaves mixed with camphor, garlic and pepper is used to treat edema, muscle pain and rheumatic pain, and its roots are also used to treat sexually transmitted diseases.

[0006] At present, there are many plant extracts with anti-aging effects. However, after retrieval, no research on the anti-aging aspect of Clerodendrum inerme extract has been found. Summary of the Invention

[0007] To solve the above technical problems, the present invention provides an anti-aging composition, its preparation method and application. The composition can improve cell viability and reduce the expression of cell senescence genes through the compounding of Clerodendrum inerme extract and Valeriana officinalis root extract, and has a significant anti-aging effect.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] The present invention provides an anti-aging composition, comprising Clerodendrum inerme extract and Valeriana officinalis root extract.

[0010] Preferably, the mass ratio of the Clerodendrum inerme extract to the Valeriana officinalis root extract is 3-10:1.

[0011] Preferably, the Clerodendrum inerme extract is an alcohol extract of at least one of the roots, stems and leaves of Clerodendrum inerme.

[0012] Preferably, the Valeriana officinalis root extract is an alcohol extract of Valeriana officinalis root.

[0013] Further preferably, the preparation method of the Clerodendrum inerme extract or the Valeriana officinalis root extract comprises the following steps:

[0014] (1) Crushing Clerodendrum inerme or Valeriana officinalis root to obtain powder;

[0015] (2) Adding an ethanol aqueous solution to the powder obtained in step (1), refluxing to obtain an extract;

[0016] (3) Freeze-drying the extract obtained in step (2) to obtain the Clerodendrum inerme extract or the Valeriana officinalis root extract.

[0017] Preferably, the particle size of the powder in step (1) is 60-120 mesh.

[0018] Preferably, the material-liquid ratio (g / 100ml) of the powder to the ethanol aqueous solution in step (2) is 1:10-15.

[0019] Preferably, the volume percentage of the ethanol aqueous solution in step (2) is 60-80%.

[0020] Preferably, the reflux in step (2) is: refluxing at 70-85°C for 1.5-4h.

[0021] The present invention also provides a preparation method of the above anti-aging composition. Specifically, the preparation method is: mixing the formula amount of Clerodendrum inerme extract and Valeriana officinalis root extract.

[0022] The present invention also provides the application of the above anti-aging composition in the preparation of cosmetics.

[0023] The present invention also provides an anti-aging cosmetic, which is made of the above anti-aging composition or the anti-aging composition prepared by the above preparation method.

[0024] The beneficial effects of the present invention are as follows:

[0025] (1) By combining the extracts of Clerodendrum inerme and Valeriana officinalis roots, the composition of the present invention can synergistically enhance the effect, improve cell viability, reduce the expression of cell senescence genes, and has a significant anti-aging effect.

[0026] (1) It was found through testing that the combination of the extract of Clerodendrum inerme and the extract of Valeriana officinalis roots of the present invention can also reduce the expression of β-galactosidase in HSF cells induced by D-galactose and delay cell senescence.

[0027] (2) The composition of the present invention is a pure plant extract, with good safety, simple preparation method and low cost, and is suitable for industrial promotion. Description of the Drawings

[0028] Figure 1 Effects of the examples and comparative examples on hydrogen peroxide-induced cell senescence and death.

[0029] Figure 2 Effects of the examples and comparative examples on hydrogen peroxide-induced cell senescence gene expression.

[0030] Figure 3 Effects of the examples of the present invention on the expression of β-galactosidase in HSF cells induced by D-galactose. Detailed Embodiments

[0031] The following describes the embodiments of the present invention in combination with specific specific examples. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0032] Before further describing the specific embodiments of the present invention, it should be understood that the protection scope of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the embodiments of the present invention are for the purpose of describing specific embodiments, rather than limiting the protection scope of the present invention.

[0033] The present invention does not limit the sources of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are all ordinary commercially available products in the technical field.

[0034] Example 1 Anti-aging Composition

[0035] An anti-aging composition, which is composed of extracts of Clerodendrum inerme and Valeriana officinalis root with a mass ratio of 3:1.

[0036] The preparation method of the extract of Clerodendrum inerme or Valeriana officinalis root comprises the following steps:

[0037] (1) Crush the roots of Clerodendrum inerme or Valeriana officinalis root, and pass through a 60-mesh sieve to obtain a powder.

[0038] (2) According to a material-liquid ratio of 1:10, add an 80% ethanol aqueous solution to the powder obtained in step (1), and reflux at 85 °C for 1.5 h to obtain an extract.

[0039] (3) Spin-dry the extract obtained in step (2) and then freeze-dry it to obtain the extract of Clerodendrum inerme or Valeriana officinalis root.

[0040] Example 2 Anti-aging composition

[0041] An anti-aging composition, which is composed of extracts of Clerodendrum inerme and Valeriana officinalis root with a mass ratio of 10:1.

[0042] The preparation method of the extract of Clerodendrum inerme or Valeriana officinalis root comprises the following steps:

[0043] (1) Crush the roots of Clerodendrum inerme or Valeriana officinalis root, and pass through a 120-mesh sieve to obtain a powder.

[0044] (2) According to a material-liquid ratio of 1:15, add a 60% ethanol aqueous solution to the powder obtained in step (1), and reflux at 70 °C for 4 h to obtain an extract.

[0045] (3) Spin-dry the extract obtained in step (2) and then freeze-dry it to obtain the extract of Clerodendrum inerme or Valeriana officinalis root.

[0046] Example 3 Anti-aging composition and its preparation method

[0047] An anti-aging composition, which is composed of extracts of Clerodendrum inerme and Valeriana officinalis root with a mass ratio of 5:1.

[0048] The preparation method of the extract of Clerodendrum inerme or Valeriana officinalis root comprises the following steps:

[0049] (1) Crush the roots of Clerodendrum inerme or Valeriana officinalis root, and pass through an 80-mesh sieve to obtain a powder.

[0050] (2) According to a material-liquid ratio of 1:12, add a 75% ethanol aqueous solution to the powder obtained in step (1), and reflux at 80 °C for 2.5 h to obtain an extract.

[0051] (3) Spin-dry the extract obtained in step (2) and then freeze-dry it to obtain the extract of Clerodendrum inerme or Valeriana officinalis root.

[0052] Example 4 Anti-aging Composition

[0053] The difference between this example and Example 3 is only that the anti-aging composition consists of extracts of Clerodendrum inerme and Valeriana officinalis root in a mass ratio of 7:1.

[0054] Example 5 Anti-aging Composition

[0055] The difference between this example and Example 3 is only that in the anti-aging composition, the extract of Clerodendrum inerme uses the leaves of Clerodendrum inerme during the preparation process.

[0056] Example 6 Anti-aging Composition

[0057] The difference between this example and Example 3 is only that in the anti-aging composition, the extract of Clerodendrum inerme uses the stems of Clerodendrum inerme during the preparation process.

[0058] Comparative Example 1 Anti-aging Composition

[0059] The difference between this comparative example and Example 3 is only that the anti-aging composition is the extract of Clerodendrum inerme.

[0060] Comparative Example 2 Anti-aging Composition

[0061] The difference between this comparative example and Example 3 is only that the anti-aging composition is the extract of Valeriana officinalis root.

[0062] By investigating the effect of the anti-aging composition on hydrogen peroxide-induced cell senescence, the anti-aging effect of the present invention was evaluated.

[0063] 1. Protective effect on hydrogen peroxide-induced cell senescence and death

[0064] HaCat cells were cultured in DMEM medium containing 10% fetal bovine serum (DMEM + 10% FBS + 1% P-S), and placed in an incubator at 37°C and 5% carbon dioxide. When the cells were in the logarithmic growth phase, the cells were seeded into 96-well plates, 100 μL per well, and the cell number was adjusted to 10,000 cells / well. Each group was set with 6 parallel wells and incubated overnight. The next day, 50 μL of the anti-aging composition solution (12.5 μg / mL, solvent was DMSO) was added to each well, and the control group was added with an equal amount of DMSO solvent. After 1 hour, 400 μM hydrogen peroxide was added to each well and incubated for another 4 hours, then 10 μL of CCK-8 reagent (Dojindo Laboratories, Japan) was added and incubated for 2 hours, and the absorbance was measured with a microplate reader at 450 nm.

[0065] Cell viability (%) = (OD of composition / OD of control) × 100%

[0066] The results are as Figure 1 shown. As can be seen from Figure 1It can be seen that before modeling, there was no significant difference in cell viability among groups compared with the control group (P>0.05); after modeling, there was a highly significant difference in cell viability in the control group compared with that before modeling ( aa P<0.01); it is indicated that the cells induced by hydrogen peroxide showed senescence and death, and the cell viability decreased significantly.

[0067] There were significant differences in cell viability among the groups of Examples 1 to 6 compared with the control group after modeling ( b P<0.05), among which, there were highly significant differences in cell viability in the groups of Examples 3, 5 and 6 ( bb P<0.01); it is indicated that the anti-aging composition of the present invention has a protective effect on hydrogen peroxide-induced cell senescence, and the effects of Examples 3, 5 and 6 are better.

[0068] The cell viability of the groups of Comparative Examples 1 and 2 after modeling showed highly significant differences compared with that before modeling ( aa P<0.01), and there was no significant difference in cell viability compared with the control group after modeling. It is indicated that at a dose of 12.5 μg / mL, the single extract of Clerodendrum inerme or the extract of Valeriana officinalis has no protective effect on hydrogen peroxide-induced cell senescence.

[0069] 2. Reduce the expression of genes related to hydrogen peroxide-induced cell senescence

[0070] After the cells were treated and administered as above, TriZol RNA extraction reagent (invitrogen, USA) was used to extract cell RNA, and total RNA was extracted and purified according to the operating steps of the Super total RNA extraction kit manual. The RNA concentration was measured using NanoDrop2000. Using the total RNA as a template, controlling the total amount of RNA in the system to be about 1 μg, it was reverse transcribed into cDNA according to the steps of the ReverTra AceqPCR RT Master Mix reverse transcription kit manual. The cDNA template obtained by reverse transcription was diluted 10 times to 100 μL, and qPCR experiments were carried out using the diluted cDNA as a template. The primer sequences of related genes are as follows:

[0071]

[0072] The expression results of the MMP-1 gene are as Figure 2 shown. It can be seen from Figure 2 that compared with before modeling, there was a significant difference in the expression level of the MMP-1 gene in the control group after modeling ( aa P<0.01), indicating that the cells induced by hydrogen peroxide can promote the expression of the MMP-1 gene.

[0073] Compared with the control group after modeling, there were extremely significant differences in the MMP-1 gene expression levels in the groups of Examples 1 to 6 after modeling ( bb P < 0.01), indicating that the anti-aging compositions provided in Examples 1 to 6 of the present invention can inhibit the MMP-1 gene expression induced by hydrogen peroxide and delay cell aging.

[0074] Compared with the control group after modeling, there was a significant difference in the MMP-1 gene expression level in Comparative Example 1 after modeling ( b P < 0.05), and there was no significant difference in the MMP-1 gene expression level in Comparative Example 2 after modeling (P > 0.05), indicating that at a dose of 12.5 μg / mL, the single extract of Clerodendrum inerme can inhibit the MMP-1 gene expression induced by hydrogen peroxide; while the single extract of Valeriana officinalis cannot inhibit the MMP-1 gene expression induced by hydrogen peroxide.

[0075] 3. Reduce the expression of β-galactosidase in HSF cells induced by D-galactose

[0076] Human skin fibroblasts HSF were cultured in DMEM medium containing 15% fetal bovine serum (DMEM + 15% FBS + 1% P-S) and placed in an incubator at 37°C with 5% carbon dioxide. In a 96-well plate, a normal control group, a model group, and an extract group (the anti-aging composition prepared in Example 2) were set up. After the model group and the extract group were cultured with complete culture medium containing 20 mg / mL D-galactose for 24 h, the extract group was added with the drug (12.5 μg / mL, and the solvent was DMSO), and the model group was added with an equal amount of DMSO solvent, and then continued to be cultured in the complete culture medium for 24 h.

[0077] Staining was performed according to the instructions of the Cellular Senescence β-Galactosidase Staining Kit (Shanghai Beyotime Biotechnology Co., Ltd.), and observations and photographs were taken under an inverted microscope. The results are as Figure 3 shown. It was found that compared with the model group, after adding the anti-aging composition of the present invention, the number of β-galactosidase-stained positive cells in the extract group decreased, indicating that the anti-aging composition of the present invention can delay cell aging.

[0078] It should be emphasized that the examples described in the present invention are illustrative rather than restrictive. Therefore, the present invention includes but is not limited to the examples described in the specific embodiments. Any other embodiments obtained by those skilled in the art according to the technical solutions of the present invention also fall within the scope of protection of the present invention.

Claims

1. An anti-aging composition, characterized in that, They are Clerodendrum inerme extract and Valeriana officinalis root extract, and the mass ratio of the Clerodendrum inerme extract to the Valeriana officinalis root extract is 3 - 10:1; the Clerodendrum inerme extract is an extract of at least one of the roots, stems and leaves of Clerodendrum inerme; The preparation method of the Clerodendrum inerme extract or Valeriana officinalis root extract comprises the following steps: (1) Crushing Clerodendrum inerme or Valeriana officinalis root to obtain powder; (2) Adding an ethanol aqueous solution to the powder obtained in step (1), refluxing to obtain an extract; (3) Freeze-drying the extract obtained in step (2) to obtain the Clerodendrum inerme extract or Valeriana officinalis root extract; The volume percentage of the ethanol aqueous solution in step (2) is 60 - 80%.

2. The composition according to claim 1, wherein The refluxing in step (2) is: refluxing at 70 - 85 °C for 1.5 - 4 h.

3. A method for preparing the anti-aging composition according to any one of claims 1-2, characterized in that, The preparation method is: mixing the Clerodendrum inerme extract and Valeriana officinalis root extract in the formula amount.

4. The application of the anti-aging composition according to any one of claims 1 - 2 or the anti-aging composition prepared by the preparation method according to claim 3 in the preparation of cosmetics.

5. An anti-aging cosmetic, which is made of the anti-aging composition according to any one of claims 1 - 2 or the anti-aging composition prepared by the preparation method according to claim 3.

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