A plant composition for delaying scalp aging and a method of preparing the same

Abstract

The present application belongs to the technical field of cosmetics, and particularly relates to a plant composition for delaying scalp aging and a preparation method thereof. The plant composition for delaying scalp aging is composed of ginseng root extract and high mountain flax flower / leaf / stem extract, wherein the mass ratio of the high mountain flax flower / leaf / stem extract to the ginseng root extract is 1:(5-60). The composition can synergistically inhibit the release of key inflammatory factors and mediators, effectively relieve the chronic inflammatory state of the scalp, and at the same time, reduce the activity of aging-related enzymes, up-regulate the expression of growth factors, and target intervention on the two key aging markers of 'chronic inflammation' and 'cell aging', so as to delay the aging of the scalp.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a plant composition for delaying scalp aging and its preparation method. Background Technology

[0002] In recent years, aging research has moved from describing phenomena to elucidating mechanisms. The authoritative review published in Cell, "The Hallmarks of Aging: An Expanding Universe," systematically updated and expanded the fourteen core biological markers of aging, providing a unified theoretical framework for understanding tissue and organ aging. Among them, "Chronic Inflammation" and "Cellular Senescence" were explicitly listed as two interrelated key markers driving the decline of multiple tissue functions.

[0003] In the scalp and skin microenvironment, chronic inflammation (also known as "inflammatory senescence") manifests as the sustained low-level release of pro-inflammatory factors (such as IL-1β, IL-6, and TNF-α), which not only disrupts the homeostasis of hair follicle stem cell niches but also accelerates collagen degradation and weakens barrier function, leading to scalp sensitivity, itching, hair loss, and an aged appearance. Simultaneously, the accumulation of cellular senescence further amplifies inflammatory signals through the senescence-associated secretory phenotype (SASP). Furthermore, these two markers form a vicious cycle in the scalp aging process: inflammation induces cellular senescence, and senescent cells, in turn, exacerbate local inflammation through SASP.

[0004] Based on the latest theoretical framework of the fourteen major aging markers, developing a novel multi-target, synergistic composition that synergistically inhibits the expression of key inflammatory factors and significantly reduces the activity of aging-related enzymes, thereby effectively blocking the vicious cycle in the scalp aging process, is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0005] The purpose of this section is to outline some aspects of the embodiments of the present invention and to briefly describe some preferred embodiments.

[0006] As one aspect of the present invention, the present invention provides a plant composition for delaying scalp aging, characterized in that the plant composition for delaying scalp aging is composed of ginseng root extract and alpine flax flower / leaf / stem extract, wherein the mass ratio of the alpine flax flower / leaf / stem extract to the ginseng root extract is 1:5-60.

[0007] As a preferred embodiment of the plant composition described in this invention, the mass ratio of the alpine flax flower / leaf / stem extract to the ginseng root extract is 1:6-30; the plant composition for delaying scalp aging is used to prepare cosmetics with the effects of delaying scalp aging and anti-inflammatory properties.

[0008] As a preferred embodiment of the plant composition described in this invention, the ginseng root extract is obtained by extraction using an aqueous solution of 1,3-butanediol as a solvent; the volume fraction of the aqueous solution of 1,3-butanediol is 10-30%.

[0009] As a preferred embodiment of the plant composition described in this invention, the volume fraction of the 1,3-butanediol aqueous solution is 20%.

[0010] As a preferred embodiment of the plant composition described in this invention, the alpine flax flower / leaf / stem extract is obtained by extraction using an aqueous ethanol solution as a solvent; the volume fraction of the aqueous ethanol solution is 60-90%.

[0011] As a preferred embodiment of the plant composition described in this invention, the volume fraction of the ethanol aqueous solution is 70%.

[0012] The present invention also provides a method for preparing the plant composition for delaying scalp aging, wherein the method for preparing the ginseng root extract includes: pulverizing ginseng root into 40-100 mesh fine powder, mixing it with a 1,3-butanediol aqueous solution with a volume fraction of 10-30% at a mass ratio of 1:5-15, extracting it at 60-90℃ for 3-6 hours, and then filtering and collecting the filtrate.

[0013] The preparation method of the alpine flax flower / leaf / stem extract includes: pulverizing alpine flax flower / leaf / stem into 40-100 mesh fine powder, mixing it with an ethanol aqueous solution with a volume fraction of 60-90% at a mass ratio of 1:5-20, heating and refluxing at 60-90℃ for 1-3 hours, combining the extracts, distilling under reduced pressure, concentrating and drying.

[0014] As a preferred embodiment of the preparation method of the plant composition for delaying scalp aging described in this invention, the ginseng root extract is prepared at an extraction temperature of 70°C for 4 hours and a material-to-liquid ratio of 1:10.

[0015] As a preferred embodiment of the preparation method of the plant composition for delaying scalp aging described in this invention, in the preparation method of the ginseng root extract, the filtration includes sequentially using a filter cloth, a 5μm filter membrane, and a 0.22μm filter membrane.

[0016] As a preferred embodiment of the preparation method of the plant composition for delaying scalp aging described in this invention, in the preparation method of the alpine flax flower / leaf / stem extract, the extraction temperature is 80℃, the extraction time is 2 hours, and the material-liquid ratio is 1:10.

[0017] The beneficial effects of this invention are as follows: Based on the core mechanism of aging, this invention uses a combination of ginseng root and alpine flax flowers / leaves / stems. After extraction through a specific process, the two produce a significant synergistic effect within a specific mass ratio range. This combination can block the scalp aging process at multiple targets. Experiments have shown that this composition can synergistically inhibit the release of key inflammatory factors (such as IL-6) and significantly reduce the production of inflammatory mediators (such as NO), effectively breaking the vicious cycle of "inflammatory aging" in the scalp microenvironment and relieving scalp sensitivity, itching, and other problems from the root cause.

[0018] The composition of this invention not only has anti-inflammatory properties, but also effectively promotes the expression of vascular endothelial growth factor (VEGF) in hair follicle-related cells (such as human dermal papilla cells), which helps improve scalp microcirculation, provides sufficient nutritional support for hair growth, and maintains the homeostasis of hair follicle stem cell niches.

[0019] Furthermore, the composition of the present invention can significantly reduce UVB-induced cellular senescence-related β-galactosidase activity, effectively alleviating the cellular senescence process caused by external factors (such as photoaging), demonstrating its potential in intervening in "cellular senescence". Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below, wherein:

[0021] Figure 1 The results are from the β-galactosidase staining experiment. Detailed Implementation

[0022] To make the above-mentioned objectives, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to specific examples.

[0023] Ginseng (Panax ginseng) is a perennial herb belonging to the genus Panax in the family Araliaceae, and is known as the "King of Herbs." Its roots are large, cylindrical or spindle-shaped, resembling a human figure, hence the name "ginseng." The main active components of ginseng include ginsenosides, polysaccharides, polyphenols, volatile oils, and various trace elements. Ginsenosides are the most abundant and structurally complex chemical components in ginseng, possessing a wide range of pharmacological effects, including enhancing immunity, anti-fatigue, nerve regulation, and metabolic regulation.

[0024] The preparation process of ginseng root extract in this invention is as follows: Fresh ginseng slices are dried to obtain ginseng slices, which are then ground into 40-100 mesh fine powder. The ginseng powder is mixed with a 1,3-butanediol aqueous solution with a volume fraction of 10-30% at a material-to-liquid ratio of 1:5-15 (mass ratio). The mixture is extracted in a water bath at 60-90℃ for 3-6 hours. Then, the extract is coarsely filtered using a 1000-mesh filter cloth to remove large particulate impurities. Finally, the extract is filtered through 5μm and 0.22μm filter membranes to collect the clear filtrate, thus obtaining the ginseng root extract.

[0025] Alpine flax (Linum alpinum) is a perennial herbaceous plant belonging to the genus Linum in the family Linaceae. It is native to the high-altitude regions of central and southeastern Europe (such as the Alps and Carpathian Mountains), and grows on rocky slopes, meadows, or forest edges at altitudes of 1,000-2,500 meters.

[0026] The preparation process of the alpine flax flower / leaf / stem extract in this invention is as follows: the alpine flax is dried and pulverized into a fine powder of 40-100 mesh. Then, the powder is mixed with an ethanol aqueous solution of 60-90% by volume at a ratio of 1:5-20 (mass ratio). The mixture is heated and refluxed at 60-90℃ for 1-3 hours. The extracts are combined and then distilled under reduced pressure to concentrate and dry.

[0027] Example 1:

[0028] Preparation of plant extracts:

[0029] Preparation of water-extracted ginseng root extract: Grind ginseng slices into 60-mesh fine powder, mix 10g of ginseng powder with 100mL of deionized water, heat and stir in a 70℃ water bath for 4 hours, coarsely filter the extract with a 1000-mesh filter cloth to remove large particulate impurities, and then filter through 5μm and 0.22μm filter membranes to obtain a clear filtrate.

[0030] Preparation of ginseng root extract by alcohol extraction: Ginseng slices were ground into 60-mesh fine powder. 10g of ginseng powder was mixed with 100mL of 10% and 20% 1,3-butanediol aqueous solutions, respectively. The mixture was heated and stirred in a water bath at 70℃ for 4 hours. The extract was coarsely filtered through a 1000-mesh filter cloth to remove large particulate impurities. Then, it was filtered through 5μm and 0.22μm filter membranes to obtain a clear filtrate.

[0031] Preparation of extracts from alpine flax flowers, leaves and stems: Alpine flax (the whole above-ground part) was dried and pulverized into 60-mesh fine powder. The powder was then mixed with a 70% ethanol aqueous solution at a ratio of 1:10. The mixture was heated and refluxed at 80°C for 2 hours. The extracts were combined and then distilled under reduced pressure to concentrate and dry.

[0032] Assay for inhibiting the expression of inflammatory cytokine IL-6:

[0033] 1. Cytotoxicity test:

[0034] RAW264.7 cells in the logarithmic growth phase with approximately 90% confluence were collected, and a cell suspension was prepared. These cells were seeded into 96-well plates and cultured at 37°C and 5% CO2 for 48 hours. A blank control group and a sample group were set up, with 3-6 replicates in each group. The cells were cultured for another 24 hours. 100 μL of CCK-8 working solution was added to each well, and the plates were incubated in a cell culture incubator for 1-4 hours. The absorbance was measured at 450 nm using a microplate reader.

[0035] 2. Inhibition of IL-6 expression as an inflammatory cytokine:

[0036] Cell suspensions were re-prepared in 96-well plates and cultured at 37°C and 5% CO2 for 48 h. A blank control group (BC group), a model group (NC group), and a sample group were set up, with 3-6 replicates per group. The sample group contained LPS working solution (LPS concentration 0.5 μg / mL) and a non-cytotoxic sample solution, the NC group contained LPS working solution, and the BC group contained only cell culture medium. Cells were cultured for another 20-24 h after sample addition. After the treatment, the supernatant was collected, and its concentration was determined according to the IL-6 ELISA kit instructions. The inhibition rate of IL-6 was calculated.

[0037] 3. Calculation of IL-6 inhibition rate:

[0038] The IL-6 inhibition rate is calculated using the following formula:

[0039] IL-6 inhibition rate (%) = (1-C) 样品 / C NC )×100

[0040] In the formula: C 样品 The IL-6 content in the sample group during the IL-6 expression inhibition assay; C NC This refers to the IL-6 content in the NC group during the IL-6 expression inhibition assay.

[0041] The effect of different concentrations of extract on macrophage survival after 24 hours of treatment was detected by CCK-8 assay. The test results are shown in Table 1.

[0042] Table 1. Cytotoxicity test results

[0043]

[0044] Using cell viability ≥85% as the criterion for non-toxicity, the maximum non-toxic concentration of the four extracts was determined. The maximum non-toxic concentration of the extracts was selected as the highest test point, and a concentration gradient was set downwards to detect the inhibitory effect of the extracts on LPS-induced macrophage secretion of IL-6. The results are shown in Table 2.

[0045] Table 2 Results of IL-6 Inhibition Test

[0046]

[0047] The data in the table compare the sample group with the blank control group; × indicates p < 0.05, meaning the data are statistically significant. Table 2 shows that all three extracts exhibited a dose-dependent inhibitory trend. Comparing ginseng root extracts prepared by two different processes revealed that the ethanol-extracted ginseng root extract showed better inhibitory effects on IL-6 than the water-extracted ginseng root extract. Furthermore, stability results showed that the water-extracted ginseng root extract became turbid after one day at room temperature, while the ethanol-extracted ginseng root extract showed no significant abnormalities after two weeks at both room temperature and 50°C. This indicates that the ethanol-extracted ginseng root extract has better stability than the water-extracted ginseng root extract.

[0048] VEGF expression promotion assay:

[0049] 1. Cytotoxicity test:

[0050] Human dermal papilla cells (HDPCs) in the logarithmic growth phase with approximately 90% confluence were collected, and a cell suspension was prepared. These cells were then seeded into 96-well plates and cultured at 37°C and 5% CO2 for 24 hours. A blank control group and a sample group were set up, with 3-6 replicates per group. The cells were cultured for another 48 hours. 100 μL of CCK-8 working solution was added to each well, and the plates were incubated in a cell culture incubator for 1-4 hours. The absorbance was measured at 450 nm using a microplate reader.

[0051] 2. VEGF content determination test:

[0052] Cell suspensions were re-prepared in 6-well plates and cultured at 37°C and 5% CO2 for 24 h. The culture medium was discarded, and a blank control group (NC group) and a sample group were set up, with two replicates for each group. The sample group contained a non-cytotoxic sample solution, while the NC group contained only the culture medium. After adding samples, the cells were cultured for another 48 h. At the end of the treatment, VEGF levels were measured according to the ELISA kit instructions.

[0053] 3. Calculation of VEGF expression upregulation rate:

[0054] The VEGF upregulation rate is calculated using the following formula:

[0055] VEGF upregulation rate (%) = (C样品 / C NC -1)×100

[0056] In the formula: C 样品 C refers to the VEGF content of the sample group in the VEGF content determination test. NC This refers to the VEGF content in the NC group during the VEGF content determination test.

[0057] The effect of different concentrations of extract on the survival rate of HDPCs was detected by CCK-8 assay. The test results are shown in Table 3.

[0058] Table 3. Cytotoxicity test results

[0059]

[0060] Using cell viability ≥85% as the criterion for non-toxicity, the maximum non-toxic concentration of three ginseng root extracts was determined. The maximum non-toxic concentration was selected as the highest test point, and a concentration gradient was set downwards to detect the promoting effect of the extracts on VEGF secretion. The results are shown in Table 4.

[0061] Table 4 Results of VEGF Expression Level Experiment

[0062]

[0063] The data in the table are compared between the sample group and the blank control group. × indicates p < 0.05, meaning the data are statistically significant. Table 4 shows that ginseng root extracts obtained through all three extraction processes have a certain upregulation effect on VEGF expression, with the ginseng root extract extracted using 20% ​​butylene glycol as the solvent exhibiting the best effect in promoting VEGF secretion.

[0064] Tests on the synergistic inhibition of the inflammatory factor IL-6 by the composition:

[0065] Based on the test results of single extracts, an extract of flaxseed flowers / leaf / stem and a ginseng root extract extracted with 20% butylene glycol were selected for combination, and the combined inhibitory effect of the two extracts on IL-6 was further studied. First, the macrophage toxicity of different concentration ratios of the combination was evaluated, and safe concentration combinations were screened based on macrophage toxicity results (cell viability ≥85%). Then, the inhibitory effect of these concentrations on LPS-induced IL-6 secretion was tested. The inhibitory effect of the combined use of flaxseed flower / leaf / stem extract and ginseng root extract on IL-6 was analyzed using the Bliss independent model. The Combination Index (CI) was calculated to evaluate the effect of the two extract combinations. The specific formula for calculating CI is as follows:

[0066] E Bliss =EA +E B -E A *E B

[0067] CI=E Bliss / E AB

[0068] In the formula: E Bliss E refers to the expected additive effect of combining drug A and drug B; A The effect of drug A when used alone; E B The effect of drug B when used alone; E AB CI refers to the actual effect of combining drug A and drug B. If CI < 1, the drug combination is considered to have a synergistic effect; if CI = 1, the drug combination is considered to have an additive effect; if CI > 1, the drug combination is considered to have an antagonistic effect.

[0069] The calculation results of macrophage toxicity, IL-6 inhibition and synergistic effect of the compound composition are shown in Table 5.

[0070] Table 5. Results of IL-6 inhibition assay using the combination of two extracts.

[0071]

[0072] As shown in Table 5, the CI values ​​of the combined use of alpine flax flower / leaf / stem extract and ginseng root extract in the mass ratio range of (1:6) to (1:30) were all less than 1, indicating a synergistic inhibitory effect on IL-6.

[0073] Composition inhibits inflammatory mediator NO test:

[0074] 1. NO Inhibition Test

[0075] The cell suspension was re-prepared in 96-well plates (RAW264.7 cells) and cultured at 37℃ and 5% CO2 for 48 h. A blank control group (BC group), a model group (NC group), and a sample group were set up, with 3–6 replicates in each group. The sample group contained LPS working solution (LPS concentration 0.5 μg / mL) and a non-cytotoxic sample solution, the NC group contained LPS working solution, and the BC group contained only cell culture medium. After adding samples, the cells were cultured for another 20–24 h. After the reaction, the supernatant was mixed with Gliese working solution and cultured in the dark for approximately 10 min, and the OD value was measured at 540 nm.

[0076] 2. Calculation of NO relative inhibition rate

[0077] The relative inhibition rate of NO is calculated according to the following formula:

[0078] NO relative inhibition rate (%) = (1-OD样品 / OD NC )×100

[0079] Where: OD 样品 OD refers to the OD value of the sample group in the Griess test; NC This refers to the OD value of the NC group in the Griess test.

[0080] The calculated NO inhibition effect of the compound composition is shown in Table 6.

[0081] Table 6. Results of NO inhibition test of the two extracts combined.

[0082]

[0083] In the LPS-induced macrophage inflammation model, LPS induces macrophages to release the inflammatory mediator NO. Excessive NO promotes the release of inflammatory factors such as IL-6 and TNF-α, which in turn promote the secretion of even more NO, creating a vicious cycle and exacerbating the inflammatory response. As shown in Table 6, the combined extracts of flaxseed flower / leaf / stem and ginseng root extract significantly inhibited the release of the inflammatory mediator NO, exhibiting multi-target anti-inflammatory effects.

[0084] β-galactosidase staining test:

[0085] 1. Cytotoxicity test

[0086] HDPCs cells in the logarithmic growth phase with approximately 90% confluence were collected, and a cell suspension was prepared. These cells were then seeded into 96-well plates and cultured at 37°C and 5% CO2 for 24 hours. A blank control group and a sample group were set up, with 3–6 replicates in each group. The cells were cultured for another 48 hours. 100 μL of CCK-8 working solution was added to each well, and the plates were incubated in a cell culture incubator for 1 hour. The absorbance was measured at 450 nm using a microplate reader.

[0087] 2. β-galactosidase staining test

[0088] Cell suspensions were re-prepared in 24-well plates and cultured at 37°C and 5% CO2 for 24 h. Three control groups (BC group), a model control group (NC group), and a sample group were set up. The blank group was not irradiated with UVB and 2 mL of complete culture medium was added. The model control group was irradiated with UVB and then 2 mL of complete culture medium was added. The sample group was irradiated with UVB and then 2 mL of complete culture medium containing the sample was added. Each group was divided into two replicates, and the cells were cultured for a further period after adding the sample. After culture, the cell culture medium was discarded, and the cells were stained according to the β-galactosidase staining kit instructions. The staining was then observed under a regular optical microscope.

[0089] After determining the effective concentration range for non-cytotoxicity using the CCK-8 assay, a UVB-induced senescence model of human dermal papilla cells was established. The senescence level of cells in each group was detected using β-galactosidase staining. Typical staining images are shown below. Figure 1 As shown. Figure 1 The β-galactosidase staining results showed that after UVB irradiation, the model group cells showed obvious blue staining, indicating that the photoaging cell model was successfully established. The staining intensity of HDPCs cells treated with the composition of the present invention was lower than that of the model group, indicating that the activity of β-galactosidase related to cell senescence was reduced. The composition of the present invention can alleviate UVB-induced cell senescence.

[0090] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A plant-based composition for delaying scalp aging, characterized in that, The plant composition for delaying scalp aging consists of ginseng root extract and alpine flax flower / leaf / stem extract, wherein the mass ratio of the alpine flax flower / leaf / stem extract to the ginseng root extract is 1:(5-60).

2. The plant composition according to claim 1, characterized in that, The mass ratio of the alpine flax flower / leaf / stem extract to the ginseng root extract is 1:(6-30); the plant composition for delaying scalp aging is used to prepare cosmetics with the effects of delaying scalp aging and anti-inflammation.

3. The plant composition according to claim 1 or 2, characterized in that, The ginseng root extract was obtained by extraction using an aqueous solution of 1,3-butanediol as a solvent; the volume fraction of the aqueous solution of 1,3-butanediol was 10-30%.

4. The plant composition according to claim 3, characterized in that, The volume fraction of the 1,3-butanediol aqueous solution is 20%.

5. The plant composition according to claim 1 or 2, characterized in that, The alpine flax flower / leaf / stem extract was obtained by extraction using an ethanol aqueous solution as a solvent; the volume fraction of the ethanol aqueous solution was 60-90%.

6. The plant composition according to claim 5, characterized in that, The volume fraction of the ethanol-water solution is 70%.

7. The method for preparing the plant composition for delaying scalp aging according to claim 1, characterized in that, The preparation method of the ginseng root extract includes: pulverizing ginseng root into 40-100 mesh fine powder, mixing it with 1,3-butanediol aqueous solution with a volume fraction of 10-30% at a mass ratio of 1:5-15, extracting it at 60-90℃ for 3-6 hours, and then filtering and collecting the filtrate. The preparation method of the alpine flax flower / leaf / stem extract includes: pulverizing alpine flax flower / leaf / stem into 40-100 mesh fine powder, mixing it with an ethanol aqueous solution with a volume fraction of 60-90% at a mass ratio of 1:5-20, heating and refluxing at 60-90℃ for 1-3 hours, combining the extracts, distilling under reduced pressure, concentrating and drying.

8. The method for preparing the plant composition for delaying scalp aging according to claim 7, characterized in that, In the preparation method of the ginseng root extract, the extraction temperature is 70℃, the extraction time is 4 hours, and the material-to-liquid ratio is 1:

10.

9. The method for preparing the plant composition for delaying scalp aging according to claim 8, characterized in that, In the preparation method of the ginseng root extract, the filtration includes sequentially using a filter cloth, a 5μm filter membrane, and a 0.22μm filter membrane.

10. The method for preparing the plant composition for delaying scalp aging according to claim 7, characterized in that, In the preparation method of the alpine flax flower / leaf / stem extract, the extraction temperature is 80℃, the extraction time is 2 hours, and the material-liquid ratio is 1:10.

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