Antioxidant plant composition for relieving alopecia and promoting hair growth

Abstract

The invention provides an antioxidant plant composition for relieving alopecia and promoting hair growth. According to the plant-derived composition disclosed by the invention, a remarkable treatment result is realized through mutual synergy and synergism, and a complementary comprehensive effect is formed. More importantly, the diversified plant components also have excellent effects of scavenging free radicals and resisting oxidation. The innovative combination provides a brand-new, efficient and multifunctional solution for preventing hair loss and promoting hair growth.

Description

Technical Field

[0001] The present invention belongs to the technical field of daily chemical products. More specifically, the present invention relates to an antioxidant plant composition for relieving hair loss and promoting hair growth. Background Art

[0002] Hair loss and scalp health problems are becoming increasingly common worldwide, seriously affecting people's quality of life and self-confidence. In particular, scalp health is directly related to hair growth and hair loss. Therefore, it is of great significance to develop a comprehensive solution that is both safe and effective.

[0003] Generally speaking, hair loss refers to the shedding of hair from the scalp and is caused by various factors. It can be divided into natural characteristics and acquired characteristics formed by other factors. Nowadays, the cause of hair loss is the same as the action of male hormones. There are internal factors and external factors such as mental stress and scalp peroxidized lipid accumulation in daily life. As is well known, these factors are involved in a complex way, leading to hair loss symptoms. In recent years, not only male hair loss, but also the number of female hair loss has increased due to changes in eating habits and increased stress caused by the social environment. The number of people suffering from the above-mentioned scalp and hair abnormal symptoms has also increased, and the age has become older and lower.

[0004] In recent years, with the increasing pursuit of beauty and health of people's appearance, the health and beauty of hair have become the focus of social attention. However, many people are faced with problems such as hair thinning and hair loss, which not only affect the individual's appearance self-confidence, but may also have a negative impact on their psychology. Hair growth is regulated by multiple factors, among which the growth of dermal papilla cells is a key link. The health and vitality of dermal papilla cells directly affect hair growth, quality and appearance. In past studies, some methods for promoting hair growth have been proposed, but there are still many challenges and limitations.

[0005] There are some hair growth products on the market, but their effects are uneven and often accompanied by adverse side effects. Minoxidil, Finasteride and Propecia belong to the drugs approved by the US FDA. Although they can promote hair growth to a certain extent, the potential side effects of these drugs (including but not limited to scalp irritation, sexual dysfunction, palpitations, etc.) and high costs make people very cautious about their long-term use.

[0006] For example, minoxidil treats hair loss mainly through the effect of dilating small blood vessels, which helps to increase scalp microcirculation. By improving blood supply, hair follicles can obtain more oxygen and nutrients, thus promoting hair growth. Minoxidil can prolong the growth stage of hair follicle cells (the anagen phase of the hair growth cycle), keeping it for a longer time, thereby increasing the hair growth time and length. Minoxidil may cause an increase in the volume of hair follicle cells, making them healthier and more active. This helps to prevent hair atrophy and slow down the hair loss process. Minoxidil may promote hair growth by stimulating the division of hair follicle cells and increasing the generation of new cells. Reducing the effect of male hormones: Although its exact mechanism is not clear, minoxidil is thought to reduce the negative effects of male hormones (such as dihydrotestosterone) on hair. This is particularly important for the treatment of male pattern hair loss, as male hormones are associated with this type of hair loss. Although minoxidil has an obvious effect on promoting hair growth, it may have side effects such as itchy eyes, itchy treatment area, redness or irritation, and palpitations, as well as hair growth in other parts of the body.

[0007] Another commonly used product is finasteride, whose treatment of male pattern hair loss (Androgenetic Alopecia, AGA) is related to genetics and male hormones. Among them, a male hormone, testosterone, will be converted into a more active form - dihydrotestosterone (DHT) in hair follicles. DHT has a negative impact on hair follicles in the hair growth cycle, causing them to gradually shrink, hair to become thinner, and eventually stop growing. The role of 5α-reductase: The key enzyme in this process is 5α-reductase, which promotes the conversion of testosterone to DHT. As a 5α-reductase inhibitor, finasteride blocks the conversion of testosterone to DHT by inhibiting the activity of this enzyme. Slowing down hair loss: By reducing DHT levels, finasteride slows down the adverse effects on hair follicles, enabling them to maintain a larger size, thus slowing down hair loss. However, finasteride mainly plays a role in maintaining existing hair and slowing down the hair loss process, and it cannot completely reverse the atrophy of already atrophied hair follicles. Some patients using finasteride report problems such as decreased libido. These side effects usually recover after discontinuation, but sometimes they may be persistent. Some men may experience breast tissue hyperplasia. Women should also avoid contact with finasteride, especially if they are pregnant or planning to become pregnant.

[0008] Therefore, developing other methods beneficial to promoting hair growth is urgently needed in this field. Summary of the Invention

[0009] The object of the present invention is to provide a plant composition that relieves hair loss, promotes hair growth, has good DPPH free radical scavenging ability at the biochemical level, and helps to protect scalp cells from oxidative stress damage.

[0010] In the first aspect of the present invention, there is provided an application of a composition for preparing a preparation for promoting the growth of hair follicle cells (including dermal papilla cells of hair follicles), alleviating hair loss, promoting hair growth, and antioxidation; the composition includes: grape (VITIS VINIFERA) fruit extract, scutellaria (SCUTELLARIA BAICALENSIS) root extract, mulberry (MORUS ALBA) root extract, saxifraga sarmentosa extract, and scutellaria alpina extract; wherein, the ratio of grape fruit extract: scutellaria root extract: mulberry root extract: saxifraga sarmentosa extract: scutellaria alpina extract is (0.004 - 0.05):(0.001 - 0.015):(0.001 - 0.015):(0.0002 - 0.004):(0.0015 - 0.02).

[0011] In another aspect of the present invention, there is provided a composition for promoting the growth of hair follicle cells, alleviating hair loss, promoting hair growth, and antioxidation, including: grape fruit extract, scutellaria root extract, mulberry root extract, saxifraga sarmentosa extract, and scutellaria alpina extract; wherein, the ratio of grape fruit extract: scutellaria root extract: mulberry root extract: saxifraga sarmentosa extract: scutellaria alpina extract is (0.004 - 0.05):(0.001 - 0.015):(0.001 - 0.015):(0.0002 - 0.004):(0.0015 - 0.02).

[0012] In another aspect of the present invention, there is provided a method for preparing a composition for promoting the growth of hair follicle cells, alleviating hair loss, promoting hair growth, and antioxidation, including: mixing grape fruit extract, scutellaria root extract, mulberry root extract, saxifraga sarmentosa extract, and scutellaria alpina extract; wherein, the ratio of grape fruit extract: scutellaria root extract: mulberry root extract: saxifraga sarmentosa extract: scutellaria alpina extract is (0.004 - 0.05):(0.001 - 0.015):(0.001 - 0.015):(0.0002 - 0.004):(0.0015 - 0.02).

[0013] In one or more embodiments, the ratio of grape fruit extract: scutellaria root extract: mulberry root extract: saxifraga sarmentosa extract: scutellaria alpina extract is (0.005 - 0.04):(0.0012 - 0.01):(0.0012 - 0.01):(0.0003 - 0.003):(0.002 - 0.015).

[0014] In one or more embodiments, the grape fruit extract: skullcap root extract: mulberry root extract: Saxifraga stolonifera extract: Scutellaria amoena extract is (0.035 ± 0.002):(0.0093 ± 0.0003):(0.0093 ± 0.0003):(0.0023 ± 0.0003):(0.014 ± 0.003).

[0015] In one or more embodiments, the concentration of the total extract in the mixed system (such as a preparation) is 0.005 - 5%, preferably 0.005 - 3%, more preferably 0.005 - 2% (for example, 0.005 - 0.2%, 0.01 - 0.15%, 0.02 - 0.12%, 0.03 - 0.1%, 0.06 - 0.08%; more specifically, such as 0.04%, 0.05%, 0.07%, 0.09%).

[0016] In one or more embodiments, the total extract exists in the mixed system (such as a preparation) in a diluted (such as diluted 1 - 50 times, specifically such as 5, 10, 20, 25, 30, 40 times) or concentrated (such as concentrated 1 - 50 times, specifically such as 5, 10, 20, 25, 30, 40 times) form (forming a mixed system).

[0017] In one or more embodiments, in the mixed system in the diluted or concentrated form:

[0018] a) Grape fruit extract, with a mass percentage of 0.01 - 10%;

[0019] b) Skullcap root extract, with a mass percentage of 0.005 - 10%;

[0020] c) Mulberry root extract, with a mass percentage of 0.005 - 10%;

[0021] d) Saxifraga stolonifera extract, with a mass percentage of 0.002 - 10%;

[0022] e) Scutellaria amoena flower / leaf / stem extract, with a mass percentage of 0.002 - 10%;

[0023] f) Dipotassium glycyrrhizinate, with a mass percentage of 0.001 - 5%;

[0024] In one or more embodiments, based on the above components, the balance is made up to 100% with a carrier (such as a cosmetic carrier), excipient or mixture to obtain a preparation.

[0025] In one or more embodiments, the total extract is mixed in Phase A, Phase B, and Phase C, wherein Phase A includes water, disodium EDTA, carbomer, sodium hydroxide (18% aqueous solution), phenoxyethanol, and ethylhexylglycerin; Phase B includes fragrance and hydrogenated castor oil; Phase C includes BLUE1 (0.1% aqueous solution); when mixing, the total extract is added to Phase A, Phase B, and Phase C to obtain a total mixture.

[0026] In one or more embodiments, the total extract is mixed in Phase A, Phase B, and Phase C, wherein Phase A includes water, grape fruit extract, disodium EDTA, sodium metabisulfite, sodium hydroxide, sodium sulfite, skullcap root extract, butylene glycol, mulberry root extract, and strawberry saxifrage extract; Phase B includes scutellaria baicalensis georgi extract, glycerin, water, and citric acid; Phase C includes dipotassium glycyrrhizinate and water; when mixing, Phase B is added to Phase A, and then Phase C is added to the mixture of Phase A and Phase B to obtain a total mixture (gel).

[0027] In one or more embodiments, the grape fruit extract is obtained by extracting grape fruits with an organic solvent (preferably ethanol); preferably, the preparation method includes: drying and crushing grape fruits, soaking them in an organic solvent (such as for 12 to 60 hours), filtering to obtain a filtrate, and removing the organic solvent (such as by vacuum evaporation); optionally, it further includes concentrating the extract (such as concentrating 2 to 50 times, specifically 5, 10, 15, 20, 30, 40 times), optionally further includes purification, and optionally further includes dilution before use (such as diluting 0.5 to 20 times, specifically 1, 2, 3, 5, 10, 15 times).

[0028] In one or more embodiments, the skullcap root extract is obtained by extracting skullcap roots with water or a water-organic solvent (preferably ethanol) mixture; preferably, the preparation method includes: drying and crushing skullcap roots, boiling them in water or a water-organic solvent mixture (such as for 30 to 120 minutes, specifically 45, 60, 75, 90, 100 minutes), filtering after cooling to obtain a filtrate, and concentrating under reduced pressure (such as concentrating 2 to 40 times, specifically 5, 10, 15, 20, 30, 35 times), and optionally further includes dilution before use (such as diluting 0.5 to 20 times, specifically 1, 2, 3, 5, 10, 15 times).

[0029] In one or more embodiments, the mulberry root extract is obtained by using mulberry roots as raw materials and extracting with an organic solvent (preferably methanol); preferably, the preparation method includes: drying and pulverizing mulberry roots, subjecting them to Soxhlet extraction with an organic solvent (such as for 5 to 12 hours, specifically 6, 8, 10, 12 hours), and filtering to obtain a filtrate; optionally, it further includes concentrating the extract (such as concentrating 2 to 50 times, specifically 5, 10, 15, 20, 30, 40 times), optionally further includes purification, and optionally further includes diluting before use (such as diluting 0.5 to 20 times, specifically 1, 2, 3, 4, 5, 10, 15 times).

[0030] In one or more embodiments, the Saxifraga stolonifera extract is obtained by using mulberry roots as raw materials and extracting with water; preferably, the preparation method includes: drying and pulverizing Saxifraga stolonifera, boiling it in water (such as for 30 to 120 minutes, specifically 45, 60, 75, 90, 100 minutes), filtering after cooling and obtaining a filtrate, concentrating under reduced pressure (such as concentrating 2 to 60 times, specifically 5, 10, 15, 20, 30, 40, 50 times), optionally further includes purification, and optionally further includes diluting before use (such as diluting 0.5 to 50 times, specifically 1, 3, 5, 10, 15, 20, 30, 40 times).

[0031] In one or more embodiments, the Scutellaria baicalensis Georgi extract is obtained by using Scutellaria baicalensis Georgi (flowers or leaves or stems or a combination thereof) as raw materials and extracting with a water-organic solvent (preferably ethanol) (70:30) mixture; preferably, the preparation method includes: drying and pulverizing Scutellaria baicalensis Georgi; soaking it in water or a water-organic solvent mixture (such as for 12 to 48 hours, specifically 18, 24, 30, 36, 42 hours), or soaking it in methanol (such as for 30 to 72 hours, specifically 36, 48, 55, 60 hours), or boiling it in a water-organic solvent mixture (such as for 30 to 120 minutes, specifically 45, 60, 75, 90, 100 minutes); filtering and obtaining a filtrate, concentrating under reduced pressure and removing the solvent (such as concentrating 2 to 40 times, specifically 5, 10, 15, 20, 30, 35 times), and optionally further includes diluting before use (such as diluting 2 to 40 times, specifically 5, 10, 15, 20, 30 times).

[0032] In one or more embodiments, the composition increases the expression of FGF-7, increases the expression of Wnt10B, increases the expression of β-catenin, decreases the expression of BMP4, and increases the DPPH free radical scavenging ability.

[0033] In one or more embodiments, the composition further includes a pharmaceutically acceptable carrier.

[0034] In one or more embodiments, each extract is in an effective amount.

[0035] In one or more embodiments, components other than the extract are adjuvants or solvents, and each adjuvant is in an effective amount.

[0036] In one or more embodiments, in the composition, grape fruit extract, Scutellaria baicalensis root extract, Morus alba root extract, Saxifraga stolonifera extract, and Scutellaria amoena extract are used as active components, or grape fruit extract, Scutellaria baicalensis root extract, Morus alba root extract, Saxifraga stolonifera extract, and Scutellaria amoena extract are used as main active components (constituting the main contribution).

[0037] In another aspect of the present invention, a method for promoting hair follicle cell growth, alleviating hair loss, promoting hair growth, and antioxidant is provided, including: administering the composition described in any one of the above to a subject in need.

[0038] In one or more embodiments, the method for promoting hair follicle cell growth, alleviating hair loss, promoting hair growth, and antioxidant is a non-therapeutic method.

[0039] In one or more embodiments, the promoting hair growth is directed to a subject with hair thinning or hair loss, and the hair thinning or hair loss is a behavior that does not belong to (does not reach) obvious disease symptoms, such as for cosmetic purposes.

[0040] In one or more embodiments, the composition further includes (but is not limited to): pH regulators, fillers, color toners (such as pigments), preservatives, stabilizers, flavoring agents, and other regulators applied in the preparation.

[0041] In one or more embodiments, the subject includes humans.

[0042] In one or more embodiments, the subject includes non-human mammals, preferably including (but not limited to): rodents (including rats, mice, hamsters, etc.), non-human primates (such as monkeys, orangutans, etc.).

[0043] In one or more embodiments, the composition is added to a cell culture medium for culturing hair follicle cells (such as expansion culture).

[0044] In another aspect of the present invention, a kit (or package (box)) for promoting hair follicle cell growth, alleviating hair loss, promoting hair growth, and antioxidant is provided, which includes the composition described above.

[0045] In one or more embodiments, in the kit, the composition is prepared as a unit-dose package / container. For example, the amount for one day is in one package / container, or the daily amount is split into separate packages / containers for several separate administrations.

[0046] Other aspects of the present invention will be apparent to those skilled in the art in view of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 The effects of plant extracts such as different amounts of grape fruits, Scutellaria baicalensis roots, Saxifraga stolonifera, etc. and the blank control group on the proliferation rate of human dermal papilla cells.

[0048] Figure 2 、 Four The effects of samples of several embodiments, samples of two comparative examples, the negative control group, and the positive control group on the expression of human FGF-7 gene; the vertical axis is "amplification multiple", which is the change multiple of the expression level of the FGF-7 gene in the experimental sample relative to the control group. That is, it indicates whether the expression of the treated sample is enhanced (up-regulated) or decreased (down-regulated) in terms of gene expression, and the degree of this change.

[0049] Figure 3 、 Four The effects of samples of several embodiments, samples of two comparative examples, the negative control group, and the positive control group on the expression of human Wnt10B gene; the vertical axis is "amplification multiple", which is the change multiple of the expression level of the Wnt10B gene in the experimental sample relative to the control group.

[0050] Figure 4 、 Four The effects of samples of several embodiments, samples of two comparative examples, the negative control group, and the positive control group on the expression of human β-catenin gene; the vertical axis "amplification multiple" refers to the change multiple of the expression level of the β-catenin gene in the experimental sample relative to the control group.

[0051] Figure 5 、 Four The effects of samples of several embodiments, samples of two comparative examples, the negative control group, and the positive control group on the activity of human BMP4; the vertical axis "amplification multiple" refers to the change multiple of the expression level of the BMP4 gene in the experimental sample relative to the control group.

[0052] Figure 6 、The calculation results of the number of shed hairs after the hair combing test for the subjects using Example 5, Comparative Example 3, and the negative control group.

[0053] Figure 7 、The local hair density test results for the subjects using Example 5, Comparative Example 3, and the negative control group.

[0054] Figure 8 The depilation test results of the subjects in Example 5, Comparative Example 3, and the negative control group were used. Detailed implementation manners

[0055] In the present invention, a composition containing plant-derived extracts is provided, which achieves significant therapeutic results through mutual synergy and enhancement. The diverse plant components form a complementary comprehensive effect, which can not only promote hair growth by activating hair growth stem cells, but also have a significant effect on treating hair loss. Using natural ingredients such as plant extracts as active ingredients greatly reduces the risk of potential side effects. This innovative combination provides a new, efficient, and multifunctional solution for preventing hair loss and promoting hair growth.

[0056] Most existing anti-hair loss products in the art usually only focus on a single treatment goal, such as only promoting hair growth, while ignoring other key needs of the scalp, such as antioxidant protection. This single nature makes consumers need to purchase multiple products to meet different needs, which is neither convenient nor cost-effective. The plant extracts of the present invention can not only effectively promote hair growth, but also have excellent antioxidant properties, providing a comprehensive and economical solution for consumers.

[0057] Term

[0058] As used herein, the term "the composition of the present invention" is a substance containing an effective amount of grape fruit extract, Scutellaria baicalensis root extract, mulberry root extract, Saxifraga stolonifera extract, and Scutellaria amoena extract as the main active ingredients (components) as described in the present invention. The weight of the active ingredients can account for 0.00001-20% of the total weight of the composition, preferably 0.0001-10%, more preferably 0.005-5%. In some required cases, dilution or concentration can also be carried out.

[0059] As used herein, the terms "comprising" or "including" include "containing", "consisting essentially of", and "consisting of".

[0060] As used herein, the term "consisting essentially of" means that in the said composition, in addition to containing the necessary ingredients or components (grape fruit extract, Scutellaria baicalensis root extract, mulberry root extract, Saxifraga stolonifera extract, and Scutellaria amoena extract), it may also contain a small amount of minor components and / or impurities that do not affect the active ingredients. For example, it may contain additives commonly used in this field or other similar fields.

[0061] As used herein, the term "pharmaceutically acceptable" component is a substance that is suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, a substance with a reasonable benefit / risk ratio.

[0062] As used herein, the term "effective amount" refers to an amount that can produce a function or activity in humans and / or animals and is acceptable to humans and / or animals.

[0063] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier used to deliver the active ingredient (effective ingredient) of the composition of the present invention into the body, including various excipients and diluents. This term refers to carriers that are not necessarily the active ingredients themselves and have no excessive toxicity after administration. Suitable carriers are well known to those of ordinary skill in the art. Auxiliary substances such as fillers, disintegrants, lubricants, glidants, effervescent agents, wetting agents or emulsifiers, flavoring agents, antioxidants, pH buffering substances, etc. may be present in these carriers.

[0064] As used herein, the term "unit dosage form" refers to a dosage form prepared for convenient administration of the composition of the present invention for a single dose, including but not limited to various gels, ointments, powders, or emulsions, etc. The unit dosage form contains the composition of the present invention suitable for a single dose, single-day, or unit-time dosage.

[0065] As used herein, "parts by weight" or "number of parts by weight" can be used interchangeably, and the parts by weight can be any fixed weight expressed in milligrams, grams, or kilograms (such as 1 mg, 1 g, 2 g, 5 g, or 1 kg, etc.). For example, a composition composed of 1 part by weight of component a and 9 parts by weight of component b can be a composition composed of 1 g of component a + 9 g of component b, or a composition composed of 10 g of component a + 90 g of component b, etc. In the said composition, the percentage content of a certain component = (the number of parts by weight of the component / the sum of the number of parts by weight of all components) × 100%. Therefore, in a composition composed of 1 part by weight of component a and 9 parts by weight of component b, the content of component a is 10%, and component b is 90%.

[0066] Composition

[0067] The present invention provides a composition for promoting hair follicle cell growth, relieving hair loss, promoting hair growth, and having antioxidant properties, including: an effective amount of grape fruit extract, Scutellaria baicalensis root extract, Morus alba root extract, Saxifraga stolonifera extract, Scutellaria amoena extract as the main activities, and they are used as the main active ingredients.

[0068] In a preferred embodiment, the grape fruit extract: Scutellaria baicalensis root extract: Morus alba root extract: Saxifraga sarmentosa extract: Scutellaria alpina extract is (0.004 - 0.05):(0.001 - 0.015):(0.001 - 0.015):(0.0002 - 0.004):(0.0015 - 0.02).

[0069] Preferably, in a specific embodiment, the composition is further prepared into a usable preparation, and the preparation comprises the following components: grape (VITIS VINIFERA) fruit extract, Scutellaria baicalensis root extract, Morus alba root extract, Saxifraga sarmentosa extract, Scutellaria alpina flower / leaf / stem extract, dipotassium glycyrrhizinate; and adjuvants / carriers: glycerol, butanediol, citric acid, sodium sulfite, sodium metabisulfite, disodium EDTA, sodium hydroxide, water.

[0070] In a preferred example, the weight percentage content of each component of the above preparation is: grape (VITIS VINIFERA) fruit extract, with a mass percentage of 0.01 - 10%; Scutellaria baicalensis root extract, with a mass percentage of 0.005 - 10%; Morus alba root extract, with a mass percentage of 0.005 - 10%; Saxifraga sarmentosa extract, with a mass percentage of 0.002 - 10%; Scutellaria alpina flower / leaf / stem extract, with a mass percentage of 0.002 - 10%; dipotassium glycyrrhizinate, with a mass percentage of 0.001 - 5%; glycerol, with a mass percentage of 1 - 20%; butanediol, with a mass percentage of 5 - 50%; citric acid, with a mass percentage of 0.001 - 5%; sodium sulfite, with a mass percentage of 0.001 - 3%; sodium metabisulfite, with a mass percentage of 0.001 - 3%; disodium EDTA; sodium hydroxide; water, the balance being made up.

[0071] In a preferred example, the above composition preparation is an aqueous solution or a spray.

[0072] The above formula range serves as a guide for the preferred solution. However, it should be understood that when used for the research and development of the preparation of the composition, the effective dose of the composition used may vary according to the actual application situation. For example, it may be made into a concentrated form or a diluted form, etc., and these variant forms should also be included in the present invention.

[0073] The present invention also includes equivalents of the active components listed in the above table, and pharmaceutical products.

[0074] There is no particular limitation on the dosage form of the composition of the present invention or the pharmaceutical preparation containing the composition, and it can be various types of dosage forms; the dosage forms that can be prepared include: solid dosage forms or liquid dosage forms, and can also be other dosage forms such as semi-solid dosage forms.

[0075] From the standpoint of easy preparation and administration, the preferred composition is a solid composition, especially a topical preparation. For example, compositions / preparations in the form of external application such as ointments or gels are preferred. Various carriers suitable for formulating external dosage forms can be applied in the present invention. As a preferred embodiment of the present invention, the external dosage form includes (but is not limited to) those selected from: ointments (such as creams, pastes), gels, foams, sprays, powders, creams, etc. Other types of dosage forms, such as powders, granules, capsules, tablets or suspensions, etc. can also be prepared.

[0076] In some preferred embodiments of the present invention, the composition is in unit dosage form. When the composition is prepared into a unit dosage form, for example, 1 dose of the unit dosage form can be administered daily / every other day; or 1 - 3 or 1 - 2 doses can be administered daily, depending on the amount of the active component in the unit dosage form.

[0077] The preparation method of the composition of the present invention is determined according to the dosage form to be prepared and the administration route.

[0078] The composition of the present invention can be added to cell culture media for the amplification culture of hair follicle cells, etc.

[0079] The compositions of the present invention are stable under accelerated conditions and they have a long shelf life (the stability is longer than three months and more than three months).

[0080] Efficacy and Applications

[0081] The present invention discloses a plant composition that relieves hair loss, promotes hair growth, has good DPPH free radical scavenging ability at the biochemical level, and helps protect scalp cells from oxidative stress damage. The composition provides a natural, safe, efficient and multifunctional anti - hair loss and scalp care solution.

[0082] The composition of the present invention promotes hair growth and hair follicle regeneration. By activating the Wnt / β - catenin signaling pathway and significantly increasing the expression levels of bone morphogenetic protein - 4 (BMP4) and fibroblast growth factor 7 (FGF7) genes, the dual promotion of hair growth and hair follicle cell regeneration is achieved, effectively promoting hair growth and the regeneration of hair follicle cells. This not only accelerates the hair growth rate, but also helps the health and vitality of hair follicle cells, providing a more comprehensive and lasting solution for anti - hair loss.

[0083] The composition of the present invention enhances the antioxidant protection of the scalp. The composition exhibits strong antioxidant properties at the biochemical level, helping to protect scalp cells from oxidative stress damage.

[0084] The composition of the present invention achieves comprehensive scalp care. In addition to its anti - hair - loss and hair - growth - promoting effects, the composition can also effectively relieve scalp inflammation, reduce dandruff and alleviate scalp itching, thus providing comprehensive scalp care.

[0085] The composition of the present invention is economical and efficient. Due to the multiple functions of the composition, consumers can avoid purchasing multiple single - function products, achieving more economical and efficient scalp and hair care.

[0086] By achieving the above - mentioned effects, the present invention not only solves multiple deficiencies of the prior art, but also provides a new and multi - functional solution with broad application prospects and commercial value.

[0087] The present invention not only discloses an effective method for preventing hair loss and promoting hair growth, but also provides a wide range of dosage form options for the comprehensive application of plant extracts through the flexible use of cosmetic processes to meet the needs of different consumers.

[0088] The combination of the present invention is composed of all - natural plant extracts, avoiding possible side effects caused by common chemical components, such as scalp irritation or allergic reactions. This feature gives the composition significant advantages for long - term use and adaptability to sensitive skin. This not only helps to maintain scalp health, but also reduces hair loss and scalp inflammation caused by oxidative stress. Therefore, in addition to the obvious anti - hair - loss and hair - growth - promoting effects, the composition also has multiple effects such as relieving scalp inflammation, reducing dandruff and alleviating scalp itching, providing a comprehensive scalp care solution.

[0089] Taking all aspects above into consideration, the present invention not only solves multiple deficiencies of the existing anti - hair - loss and scalp care technologies, but also provides a new, multi - functional, safe and efficient solution. This innovative technology has high application prospects and commercial value.

[0090] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. For the experimental methods without specific conditions noted in the following embodiments, they are generally carried out under conventional conditions such as those described in Molecular Cloning: A Laboratory Manual (4th Edition) edited by (USA) M.R. Green (Michael R. Green) and (USA) J. Sambrook (Joseph Sambrook), translated by He Fuchu (2017), published by Science Press, or according to the conditions recommended by the manufacturer. For plant extraction conditions, unless otherwise stated, they refer to the conventional operation techniques in the art.

[0091] I. Preparation of Extracts

[0092] Preparation of grape fruit extract:

[0093] Plant material: Grape fruits.

[0094] Solvent: Ethanol.

[0095] Extraction process:

[0096] 1. Wash the grape fruits, dry and crush them.

[0097] 2. Soak the crushed grape fruits in ethanol for 48 hours.

[0098] 3. Filter the mixture and collect the filtrate.

[0099] 4. Evaporate ethanol under reduced pressure to obtain a concentrated extract (the extract can be further purified, such as by column chromatography).

[0100] Concentration multiple: 10-fold concentration.

[0101] Dilution multiple: It can be diluted to 1 - 5 times according to needs before use. The grape fruit extract used in the subsequent examples is the product after diluting the concentrated extract to 1.5 times.

[0102] Preparation of Scutellaria baicalensis root extract:

[0103] Plant material: Scutellaria baicalensis roots.

[0104] Solvent: Water or water-ethanol mixture.

[0105] Extraction process:

[0106] 1. Wash the Scutellaria baicalensis roots, dry and crush them.

[0107] 2. Put the crushed Scutellaria baicalensis roots into the water-ethanol mixture and boil for 1 hour. In terms of the use of the solvent, a mixed solvent of water and ethanol (30% water and 70% ethanol by volume) can more effectively extract the water-soluble and fat-soluble components in Scutellaria baicalensis roots, ensuring the broad-spectrum activity of the extract.

[0108] 3. Filter the mixture after cooling and collect the filtrate.

[0109] 4. Concentrate the filtrate under reduced pressure to obtain the extract (the extract can be further purified, such as by recrystallization).

[0110] Concentration multiple: 8-fold concentration.

[0111] Dilution multiple: Dilute to 1 - 3 folds as needed before use. The Scutellaria root extract used in the subsequent examples was diluted to 4 folds.

[0112] Preparation of mulberry root extract:

[0113] Plant material: Mulberry roots.

[0114] Solvent: Methanol.

[0115] Extraction process:

[0116] 1. Wash the mulberry roots, dry and crush them.

[0117] 2. Perform Soxhlet extraction with methanol for 8 hours.

[0118] 3. Filter and concentrate the extract (it can be further purified using liquid partitioning or column chromatography).

[0119] Concentration multiple: 12-fold concentration.

[0120] Dilution multiple: Dilute as needed before use. The mulberry root extract used in the subsequent examples was diluted to 6 folds.

[0121] Preparation of Saxifraga stolonifera extract:

[0122] Plant material: Whole plant of Saxifraga stolonifera.

[0123] Solvent: Water.

[0124] Extraction process:

[0125] 1. Wash the Saxifraga stolonifera, dry and crush it.

[0126] 2. Boil the crushed Saxifraga stolonifera in water for 1 hour.

[0127] 3. Cool and filter the mixture and collect the filtrate.

[0128] 4. Evaporate the water under reduced pressure to obtain the concentrated extract.

[0129] 5. The extract can be further purified using column chromatography.

[0130] Concentration / dilution multiple:

[0131] Concentration multiple: 15-fold concentration.

[0132] Dilution multiple: Dilute to 3 - 5 times as needed before use. The strawberry saxifrage extract used in the subsequent examples was diluted to 30 times.

[0133] Preparation of Scutellaria baicalensis Georgi flower / leaf / stem extract:

[0134] Plant materials: Scutellaria baicalensis Georgi flowers, leaves or stems.

[0135] Solvents: Ethanol-water mixture or methanol.

[0136] Extraction process:

[0137] 1. Preparation: Wash, dry and crush the flowers, leaves and stems of Scutellaria baicalensis Georgi (equal amounts of flower / leaf / stem mixture) separately.

[0138] 2. Soaking:

[0139] Flowers: Soak in ethanol-water mixture (70:30 by volume) for 24 hours.

[0140] Leaves: Soak in methanol for 48 hours.

[0141] Stems: Boil in water-ethanol mixture for 1 hour.

[0142] 3. Filtration: Filter the soaked mixture and collect the filtrate.

[0143] 4. Concentration: Evaporate the solvent under reduced pressure to obtain the concentrated extract.

[0144] Flower extract: Concentrated ethanol (10-fold concentration).

[0145] Leaf extract: Concentrated methanol (12-fold concentration).

[0146] Stem extract: Water-ethanol mixture (15-fold concentration) (the extract can be further purified, such as by liquid-liquid partitioning or column chromatography).

[0147] For the flower / leaf / stem extracts used in the subsequent examples, dilute the above extracts to 4 times as needed before use.

[0148] Each of the above-prepared extracts is used for the preparation of the subsequent composition formulations.

[0149] 2. Preparation of composition formulations

[0150] Prepare a plant-derived anti-hair loss and / or hair growth composition preparation, see Examples 1-4 and Comparative Examples 1-2 (Table 1). The composition preparation is used to treat the proliferation rate of human dermal papilla cells in hair follicles, as well as the mRNAs of Wnt / β-catenin, bone morphogenetic protein-4 (BMP4), and fibroblast growth factor 7 (FGF7) factors in human dermal papilla cells.

[0151] Table 1

[0152]

[0153] II. The composition preparations of Examples 1-4 are composed of Processes A, B, and C

[0154] Prepare A, B, and C separately, and then mix them into a system:

[0155] Phase A: Water, VITIS VINIFERA fruit extract, disodium EDTA, sodium metabisulfite, sodium hydroxide, sodium sulfite, SCUTELLARIA BAICALENSIS root extract, butylene glycol, MORUS ALBA root extract, SAXIFRAGA SARMENTOSA extract, and mix to obtain Phase A.

[0156] Phase B: SCUTELLARIA ALPINA flower / leaf / stem extract, glycerin, water, citric acid, and mix to obtain Phase B.

[0157] Phase C: Dipotassium glycyrrhizinate, water, and mix to obtain Phase C.

[0158] During preparation:

[0159] Add the components of Phase A to the emulsifying pot in sequence, stir evenly at room temperature to obtain a mixture of Phase A components;

[0160] Add the components of Phase B to the emulsifying pot in sequence, stir evenly at room temperature to obtain a mixture of Phase A + Phase B components;

[0161] Add the components of Phase C to the emulsifying pot in sequence, stir evenly at room temperature to obtain a mixture of Phase A + Phase B + Phase C components, and discharge to obtain the total mixture.

[0162] Place the total mixture under appropriate conditions for 24 hours. During this period, sample the mixture regularly to ensure compliance with specific hygienic chemical and microbiological indicators through professional chemical and microbiological analyses. Verify that the mixture meets the required standards, inject it into the predetermined packaging bottles using a filling machine, and perform airtight packaging. Finally, store the sealed product in a cool place or in the refrigerator's freezer to ensure its quality and stability.

[0163] In Comparative Examples 1-2, the components were increased or decreased according to the formula, and the basic process remained unchanged compared to the procedures of Examples 1-4.

[0164] III. Determination of the proliferation rate of human dermal papilla cells using the MTT assay

[0165] The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay is a method commonly used to evaluate the cell proliferation rate. The steps for determining the proliferation rate of human dermal papilla cells using the MTT assay are as follows:

[0166] 1) Minoxidil was used as the positive control group, and the blank sample was used as the negative control. The samples to be tested from Examples 1-4 and Comparative Examples 1-2 were also included.

[0167] 2) Cell culture: Human dermal papilla cells were cultured under appropriate conditions until the appropriate growth stage.

[0168] 3) Cell seeding: An appropriate amount of cell suspension was added to each well of a 96-well microplate, usually 10 4 cells / well.

[0169] 4) Cell incubation: The microplate was placed in an incubator at 37°C and 5% CO 2 for 24 hours to allow the cells to attach and grow.

[0170] 5) Sample treatment: Four different concentrations of the samples to be tested were added to four groups of cells respectively. Minoxidil (500 μM) was added to the positive control group, and no treatment was added to the negative control group, only the culture medium was used.

[0171] 6) Continue incubation: Incubate for an appropriate time (e.g., 24 hours) to observe the effect of the samples.

[0172] 7) Addition of MTT solution: 10 μL to 100 μL of MTT solution (usually 5 mg / mL) was added to each well.

[0173] 8) Continue incubation: Incubate at 37°C for 1 - 4 hours to convert MTT into blue-violet formazan crystals.

[0174] 9) Stop the reaction: Remove the culture medium and add a solvent (e.g., DMSO or isopropanol) to dissolve the crystals.

[0175] 10) Measurement of absorbance: Measure the absorbance at 570 nm, with 630 nm as the reference wavelength.

[0176] 11) Data analysis: Calculate the cell proliferation rate using a specific formula.

[0177] The formula for calculating the cell proliferation rate is as follows:

[0178]

[0179] Among them,

[0180] ABSsample represents the absorbance data of the sample to be detected (with cells, with sample).

[0181] ABSblank represents the absorbance data of the blank control sample (without cells, without sample, without positive / negative control).

[0182] ABSnegative control represents the absorbance data of the negative control group (with cells, without sample).

[0183] Human dermal papilla cells play a key role in hair follicle development and growth. These cells interact with hair follicle growth. As Figure 1 shown, with the increase in the usage amount of plant extracts such as grape fruit, Scutellaria baicalensis root, and Saxifraga stolonifera in the plant composition, the proliferation rate of human dermal papilla cells has increased significantly compared to the blank control group. Starting from the extract concentration of 0.01%, the cell proliferation rate shows a trend of increasing with the increase in concentration. Specifically, when the extract concentration is 0.07%, the cell proliferation rate reaches the highest value, which is 281%, and is significantly higher than the positive control group, with particularly excellent effects.

[0184] The above results suggest that the specific extract of the present invention is a preparation that can effectively promote hair follicle cell proliferation. Except for the extracts or extract combinations in Examples 4 and 1 - 3, although their proliferation rates are lower than those of the positive control group, they are still significantly higher than the baseline control group (240%), showing their positive effects on cell proliferation. Papilla cell proliferation is usually a good indicator of increased hair growth activity. More papilla cells mean stronger signal transduction and nutrient supply, thus contributing to the rapid and healthy growth of hair.

[0185] IV. qRT-PCR analysis of key pathways for cell growth and differentiation

[0186] Use qRT-PCR to detect β-catenin, Wnt10b, FGF7, and BMP indicators.

[0187] Use qRT-PCR to detect β-catenin, Wnt10b, FGF7, and BMP indicators to accurately quantify the expression of these genes, so as to analyze the key pathways for cell growth and differentiation, and assist in disease treatment and tissue engineering analysis.

[0188] The steps for using qRT-PCR to detect β-catenin, Wnt10b, FGF7, and BMP indicators are as follows:

[0189] 1) Sample processing and condition setting: Four experimental groups were established, including a blank control (culture medium, no induction condition), a negative control (culture medium, DHT induction condition), a positive control (minoxidil, 500 μM), and a test sample (culture medium, sample, DHT induction condition).

[0190] 2) RNA extraction and cDNA synthesis: Total RNA in human dermal papilla cells was extracted by the Trizol method and converted into cDNA using a reverse transcriptase reaction system.

[0191] 3) Primer design and preparation of qRT-PCR mixture: Specific primers were designed for the β-catenin, Wnt10b, FGF7, and BMP genes, and a qRT-PCR mixture including cDNA, appropriate buffers, primers, and SYBR Green or TaqMan probe was prepared.

[0192] 4) qRT-PCR reaction: qRT-PCR was performed under appropriate thermal cycling conditions for quantitative analysis.

[0193] 5) Ct value recording: The Ct values of the target genes and the Ct values of the selected reference genes in each group were recorded, and the latter were used as internal standards to eliminate inter-sample variation.

[0194] 6) Calculate the ΔCt value, and the ΔΔCt value is used to quantify the relative change in gene expression levels.

[0195] 7) Statistical analysis and result interpretation: Appropriate statistical analysis was performed on the obtained data to determine the significant differences between groups and to interpret the possible biological significance of the expression differences of β-catenin, Wnt10b, and BMP in the growth and other processes of human dermal papilla cells.

[0196] ΔCt (Delta Ct) is the difference between the Ct value of the target gene and the Ct value of the internal reference gene. The calculation formula is as follows:

[0197] ΔCt = Ct target gcnc - Ct reference genc ;

[0198] ΔΔCt (Delta Delta Ct) is the difference between the ΔCt values of the treatment group and the control group. The calculation formula is as follows:

[0199] ΔΔCt = ΔCt experimental group - ΔCt control group ;

[0200] The relative expression of the target gene was calculated using the ΔΔCt value, and the calculation formula is as follows:

[0201] Relative Expression=2 -ΔΔCt 。

[0202] (1) FGF-7 (Fibroblast Growth Factor 7) Expression Analysis

[0203] FGF-7 (Fibroblast Growth Factor 7) plays a key role in various biological processes, especially in cell proliferation, migration, and tissue repair, where it serves as an important regulator. During hair growth, FGF-7 is considered a key regulatory factor in the interaction between dermal papilla cells and peripheral follicular cells. Its function can activate dermal papilla cells, thereby promoting hair growth and regeneration.

[0204] As Figure 2 shown, by increasing the concentrations of plant extracts such as grape fruit, Scutellaria baicalensis, and Saxifraga stolonifera in the composite plant composition, the FGF-7 gene expression was detected in six different samples, showing a significant upregulation effect, and the upregulation effect was positively correlated with the sample concentration. This indicates that the active ingredients in the samples may act as activators of FGF-7, effectively enhancing the activity of dermal papilla cells, thereby playing a promoting role in cell proliferation and hair regeneration.

[0205] Specifically, as the concentration of the plant composition gradually increases, the upregulation effect of FGF-7 gene expression gradually strengthens. In Example 1 (0.01%) to Example 4 (0.07%), the upregulation rate of FGF-7 gene expression gradually increases from 0.859 to 1.425, showing a significant effect of concentration increase on gene expression upregulation.

[0206] Especially in Example 4, the upregulation effect of FGF-7 gene expression is the most significant, reaching 1.425, indicating that under high-concentration conditions, these plant extracts have the strongest activation effect on FGF-7. In comparison, Comparative Example 1 and Comparative Example 2 are significantly lower than Example 4. This shows that in the absence of certain synergistic plant components, the overall effect of the composition will be significantly weakened.

[0207] In addition, the FGF-7 expression value shown by the negative control (NC) is only 0.624, while the positive control (minoxidil 500 μM) reaches 1.416, further verifying the effectiveness of the composition's activation effect on FGF-7.

[0208] Overall, the experimental results show that the composite composition containing multiple plant extracts (especially Example 4) is effective in activating FGF-7 gene expression, promoting the activity of dermal papilla cells, and supporting hair regeneration, while the effect of a single extract is far less than that of a combination of multiple components. This finding supports the potential application of plant compositions in hair growth and provides a scientific basis for the development of new plant extract hair care products.

[0209] (2) Wnt10B Expression Analysis

[0210] Wnt10B is an important signaling molecule in the Wnt family and is involved in various biological processes, including cell proliferation, differentiation, and migration. During hair growth, Wnt10B is considered a key factor promoting the proliferation of hair follicle cells and the activation of dermal papilla cells. It plays a crucial role in hair growth and regeneration by regulating the activity of key cells in the hair follicle.

[0211] As Figure 3 shown, the samples of Examples 1 to 4 and Comparative Examples 1 to 2 showed different degrees of upregulation levels in Wnt10B gene expression, and these upregulation effects were closely related to their potential hair growth-promoting ability. It can be seen from the figure that as the concentration of the plant extract increased, the upregulation effect of Wnt10B gene expression gradually enhanced. Especially in Example 4, the upregulation effect was the most significant, reaching 2.2371, indicating that it had the strongest potential for promoting hair growth.

[0212] This positive correlation between concentration and upregulation of gene expression further suggests that as a key regulatory molecule in hair growth, the upregulation of Wnt10B expression is closely related to the increase in the concentration of the plant composition. Therefore, the synergistic effect of multiple effective plant extracts in Example 4 may have maximally promoted the upregulation of Wnt10B, and thus is expected to significantly enhance the proliferation of hair follicle cells and the activity of dermal papilla cells, ultimately resulting in the promotion of hair growth or the effect of anti-hair loss.

[0213] On the other hand, the Wnt10B gene upregulation effects of Comparative Examples 1 and 2 were 0.8046 and 0.9074 respectively, which were significantly lower than that of Example 4. This phenomenon indicates that a single extract or a combination of some components may not be sufficient to fully activate the expression of Wnt10B, further emphasizing the significance of the synergistic effect between multiple plant extracts on gene upregulation and its biological effects.

[0214] Generally speaking, these data clearly show that the upregulation effect of Wnt10B is proportional to the concentration of the plant composition and the synergistic effect of the components. The significant upregulation effect of Wnt10B in the examples indicates that these compositions have good potential for promoting hair growth or anti-hair loss effects, especially when multiple plant components act synergistically, the effect is more significant.

[0215] (3) β-catenin Expression Analysis

[0216] β-catenin is a key regulator in the Wnt signaling pathway and plays important roles in various biological processes, including cell proliferation, differentiation, and migration. During hair growth, the activation of β-catenin usually reflects the upregulation of the Wnt signaling pathway, which is crucial for the biological activity, proliferation ability of hair follicle cells, and hair regeneration.

[0217] As Figure 4 shown, the different upregulation effects of the four example samples and the two comparative example samples on β-catenin gene expression indicate their differences in promoting the biological state of hair follicle cells. The diffusion multiple of Example 1 is 0.7594, which is relatively low, indicating that the expression level of β-catenin is low or signal transduction may be inhibited to a certain extent, which may be related to the weakening of the proliferation or differentiation ability of hair follicle cells.

[0218] With the increase in the concentration of the plant extract, the diffusion multiple of Example 2 increases to 0.7939, which may reflect the initial activation of the β-catenin signaling pathway, thus potentially leading to a gradual increase in the proliferation and differentiation ability of hair follicle cells. The diffusion multiple of Example 3 further increases to 0.8681, and the diffusion multiple of Example 4 is the highest, reaching 0.9112, indicating that as the concentration of the plant extract gradually increases, the activation effect of the β-catenin signaling pathway also enhances, thus potentially significantly promoting the activity and regeneration ability of hair follicle cells.

[0219] The diffusion multiples of Comparative Example 1 and Comparative Example 2 are 0.6799 and 0.7496 respectively, which are relatively low. Although their extract concentrations are the same as those of Example 4, due to the lack of multiple components with synergistic effects, their β-catenin activation effects are significantly inferior to those of Example 4. This indicates that the synergistic effect of multiple plant extracts is crucial for the activation effect of the Wnt signaling pathway, and the effects of single components or combinations of some components are often insufficient to significantly enhance gene expression.

[0220] In addition, the β-catenin gene expression value of the negative control (NC) is 0.6489, which is lower than that of all experimental groups, further verifying the positive effect of the plant extract on the activation of the Wnt signaling pathway. The expression value of the positive control (minoxidil 500 μM) is 0.9496, reaching the highest level, demonstrating its significant effect in promoting hair follicle cell proliferation and signal pathway activation.

[0221] In summary, the experimental results show that the plant extract combinations in Examples 1-4 can effectively upregulate the expression of β-catenin, activate the Wnt signaling pathway, and thus promote the proliferation and differentiation of hair follicle cells, contributing to hair growth and regeneration. In contrast, Comparative Examples 1 and 2, due to the lack of multiple components with synergistic effects, have significantly inferior effects compared to the Examples. These findings provide a scientific basis for the further development of plant extracts for hair growth and anti-hair loss products.

[0222] Wnt10B is a key component of the Wnt signaling pathway, which can activate β-catenin, thereby affecting cell proliferation, differentiation, and migration. The interaction between Wnt10B and β-catenin is particularly crucial in the growth and development of hair follicles.

[0223] Combined with the previous data analysis on β-catenin and Wnt10B factors, it can be understood that:

[0224] Example 1: Exhibits stable activation of the Wnt signaling pathway, which is necessary for the normal function of hair follicles but may not be sufficient to significantly promote hair follicle growth.

[0225] Example 2: A certain degree of Wnt10B activation indicates significant hair follicle proliferation potential.

[0226] Example 3: Shows further activation of Wnt10B and β-catenin, indicating a significant promoting effect on hair follicle growth and differentiation.

[0227] Example 4: A significant increase in Wnt10B may reflect strong activation of the Wnt signaling pathway. This may be related to an increase in the significant proliferation or differentiation ability of hair follicle cells, indicating that Example 4 has strong potential to promote hair follicle growth and development.

[0228] It can be seen that Example 3 (concentration 0.05%) and Example 4 (concentration 0.07%) particularly show the potential to promote hair follicle growth.

[0229] (4) BMP4 expression analysis

[0230] BMP4 (bone morphogenetic protein 4) is a member of the TGF-β (transforming growth factor-β) superfamily. It plays a key role in many biological processes, including cell proliferation, differentiation, migration, and apoptosis. The hair follicle cycle consists of three main stages in the hair growth process: anagen, catagen, and telogen. BMP4 plays a role in all three stages.

[0231] 1) Anagen

[0232] Inhibitory effect of BMP4: During the anagen phase, hair follicle cells proliferate rapidly, promoting hair growth. BMP4 generally plays an inhibitory role during this stage. It restricts the activation and differentiation of hair follicle stem cells, preventing excessive growth.

[0233] 2) Catagen

[0234] Promoting effect of BMP4: The catagen phase is a reverse stage of hair growth where hair follicle cells begin to apoptose and hair growth stops. An increase in BMP4 activity may drive the hair follicle into this stage, promoting the apoptosis and structural remodeling of hair follicle cells.

[0235] 3) Telogen

[0236] BMP4 maintains the resting state: During the telogen phase, the hair follicle is in a quiescent state and hair no longer grows. BMP4 may help maintain the resting state of hair follicle stem cells during this stage.

[0237] BMP4 plays a key role in each stage of the hair follicle cycle, affecting hair growth and regression by finely regulating the behavior of hair follicle stem cells. A decrease in BMP4 may cause the hair follicle to enter the anagen phase and promote hair growth. This mechanism may provide new therapeutic strategies for hair regeneration and alopecia treatment.

[0238] Based on the role of BMP4 in the hair follicle cycle and hair growth, by analyzing Figure 5 , it can be seen that the sample with a concentration of 0.07% has the highest percentage of BMP4 downregulation, indicating that it may be the most effective in promoting hair growth because the inhibition of BMP4 activity is reduced, which may enhance hair follicle activity; although the other concentration samples have a lower percentage of BMP4 downregulation, they may still be beneficial to hair growth, but the effect is not as significant as that of the 0.07% concentration. The biological significance of these results may point to the potential use of these extracts in regulating BMP4 activity, thus being able to be used in the treatment or products for promoting hair growth.

[0239] Analysis Figures 1 - 5 It can be known that the mixture of plant components such as Scutellaria baicalensis, Fragaria vesca, and Saxifraga stolonifera provided by the present invention can effectively activate and regenerate hair follicle cells when treating human hair follicle dermal papilla cells, with a significant activation effect on the Wnt / β-catenin signaling pathway, a significant increase in the expression levels of bone morphogenetic protein-4 (BMP4) and fibroblast growth factor 7 (FGF7) genes, and a significant proliferation in the number of hair follicle cells observed after application.

[0240] V. Analysis of DPPH free radical scavenging ability

[0241] Determine the DPPH free radical scavenging rate of the plant composition to evaluate its antioxidant capacity.

[0242] When studying the antioxidant capacity of plant compositions, the use of DPPH (1,1-diphenyl-2-picrylhydrazyl) data can provide strong support. By testing the DPPH radical scavenging ability of plant compositions, quantitative data on their antioxidant potential can be obtained. A high DPPH radical scavenging rate means that the plant composition helps to neutralize free radicals, thereby slowing down scalp aging, reducing inflammation, and promoting the growth of healthy hair. These ingredients provide an effective multi-dimensional solution for comprehensively improving scalp and hair health.

[0243] The DPPH method is used to quantitatively determine the antioxidant capacity of biological samples, classified drugs, and foods. This method is based on the fact that DPPH free radicals have a single electron and have a strong absorption at 517 nm, and its alcohol solution is purple. When an antioxidant is present, the DPPH free radicals are scavenged, the solution color fades, and the degree of fading is quantitatively related to the degree of scavenging (i.e., the change in absorbance). Therefore, rapid quantitative analysis can be carried out using a spectrophotometer or a microplate reader. The experimental steps are as follows:

[0244] 1) Prepare the DPPH solution: Prepare a DPPH ethanol solution of a certain concentration (usually 0.1 mM) for the experiment. Ensure that the solution is stored in a dark place to prevent photodegradation.

[0245] 2) Prepare the sample solution: Extract or prepare the plant composition to obtain an appropriate concentrate or solution.

[0246] ① Take a set of test tubes and add the same volume of DPPH solution to each test tube.

[0247] ② For each test tube, gradually add different concentrations of the plant composition sample so that the final concentration of each test tube is different.

[0248] ③ At the beginning of the experiment, record the absorbance baseline of the DPPH solution before adding the sample, which can be measured using a spectrophotometer.

[0249] 3) Reaction process: Let the sample react with the DPPH solution under dark conditions for a certain period of time, usually between 30 minutes and 1 hour, to ensure that the reaction proceeds fully.

[0250] 4) Absorbance measurement: Measure the absorbance of the solution in each test tube using a spectrophotometer, usually at a specific wavelength (such as 517 nm). Record the absorbance value of each test tube.

[0251] Use the following formula to calculate the DPPH radical scavenging rate of each sample:

[0252]

[0253] Among them, A0 is the absorbance value of the control sample, and A1 is the absorbance value of the sample. The control sample is a solution sample containing only DPPH.

[0254] Graph plotting and data analysis: Plot the relationship between the DPPH radical scavenging rate and the sample concentration as a graph. The IC50 value can be calculated by fitting the data. The IC50 value refers to the concentration at which the DPPH radical scavenging rate reaches 50%. To calculate the IC50 value, a non-linear regression model is usually used to fit the data. The most commonly used one is the logarithmic-response model (Logistic Function). The mathematical expression of this model is as follows:

[0255]

[0256] Among them,

[0257] y represents the DPPH radical scavenging rate (%).

[0258] x represents the concentration (μg / ml).

[0259] Max represents the maximum scavenging rate.

[0260] Hillslop represents the slope.

[0261] logIC50 is the corresponding value of IC50.

[0262] According to the classification of Blois (1958), when the IC50 value is used to evaluate the antioxidant activity of a compound, it is defined as follows:

[0263] Having very strong antioxidant activity: The IC50 value is less than 50 ppm.

[0264] Having strong antioxidant activity: The IC50 value is between 50 - 100 ppm.

[0265] Having medium antioxidant activity: The IC50 value is between 100 - 150 ppm.

[0266] Having weak antioxidant activity: The IC50 value is between 150 - 200 ppm.

[0267] Table 2 shows the test results of the DPPH radical scavenging rate of Examples 1 - 4 of the plant composition of the present invention. The positive control sample is α-tocopherol. α-tocopherol is the most active form in the tocopherol family and is widely used in antioxidant research. Its DPPH radical scavenging activity has been widely studied.

[0268] Table 2

[0269] Sample name IC50 (ppm) α-Tocopherol 11.45 Example 4 13.85 Comparative Example 1 135.21 Comparative Example 2 78.32

[0270] As can be seen from the results in Table 2, Example 4 showed a very significant DPPH radical scavenging rate.

[0271] Therefore, the composition preparation of the present invention exhibits obvious DPPH radical scavenging ability, and its activity shows a dose-dependent relationship. As the tested concentration increases, its scavenging rate of DPPH radicals shows a significant increase. This phenomenon is particularly significant within a certain concentration range. The research results clearly reveal that the developed plant composition exhibits significant antioxidant activity at the biochemical level.

[0272] VI. Preparation and Effect Analysis of Example 5 and Comparative Example 3

[0273] 1. Formulation of the Composition Preparation (Gel)

[0274] Considering that the efficacy of the drug at the biological level will decrease relative to the cellular level. Therefore, the concentration of the active ingredient of the composition in Example 4 of the present invention was amplified, and the composition preparation of Example 5 was provided to investigate its effect on the human scalp. Specifically, a refreshing gel for the scalp was prepared.

[0275] Example 5 provides a composition and preparation containing a higher dosage, which is 25 times the amplification of Example 4, and a cosmetic composition with an effective plant extract (target plant extract) at a weight ratio of 1.75%. At the same time, a cosmetic composition containing Platycladus orientalis extract was used as Comparative Example 3, and a blank composition without any plant extract was used as the negative control group (NC).

[0276] Table 3

[0277]

[0278] 2. Preparation of the Compositions of Example 5, Comparative Example 3, and Negative Control Group (NC)

[0279] The raw materials and manufacturers used in the composition formulations of Example 5, Comparative Example 3, and Negative Control Group (NC) are shown in Table 4.

[0280] Table 4

[0281]

[0282] The preparation process of this scalp gel includes the following steps:

[0283] Phase A: Dissolve disodium EDTA in water.

[0284] Phase A: Uniformly disperse carbomer into Phase A, and control the stirring speed at about 800 rpm.

[0285] Phase A: Slowly add an aqueous solution of 18% sodium hydroxide to Phase A. After neutralization, the measured pH is around 7.0 - 7.2. Stir evenly. Control the stirring speed at around 1000 rpm.

[0286] Phase A: Add the preservatives phenoxyethanol and ethylhexylglycerin.

[0287] Phase B: In a separate beaker, stir the fragrance and PEG - 40 hydrogenated castor oil evenly and add them to Phase A.

[0288] Phase C: Add colorants such as dyes to the previous separated phase.

[0289] Phase D: Add the plant extract composition and stir evenly.

[0290] The composition preparation of Example 5 obtained above is a scalp gel with a light texture and non - sticky hands. Calculated by weight ratio, the raw materials include 1.5% carbomer, 3.7% of an 18% sodium hydroxide aqueous solution, 0.30% PEG - 40 hydrogenated castor oil, 0.10% disodium EDTA, 0.72% phenoxyethanol, 0.08% ethylhexylglycerin, 0.4% of a 0.1% aqueous solution of BLUE 1, and a plant extract composition accounting for 6.8% of the total obtained by magnifying Example 4 by 10 times.

[0291] The difference between Comparative Example 3 and Example 5 is only that the plant extract composition in the formula is different. The negative control group is a blank formula without any plant extract.

[0292] According to the following evaluation methods, the scalp - refreshing gel formulations of Example 5, Comparative Example 3, and the control group were evaluated. The evaluation items include: "effect of reducing hair loss", "effect of promoting hair growth", "effect of reducing dandruff", "reduction of scalp itching condition", and "stability of the preparation".

[0293] 3. Analysis of the effect in reducing hair loss

[0294] Overall, refer to the "Test Method for Evaluating the Anti - hair Loss Efficacy of Cosmetics" in the "Technical Specifications for Cosmetics Safety (2015 Edition)" to evaluate the effect of reducing hair loss of the test composition preparation.

[0295] (1) Subject inclusion criteria

[0296] a. Healthy women aged 35 - 50.

[0297] b. Hair length between 5 - 40 cm.

[0298] c. Those who are troubled by excessive hair loss and mild hair thinning, and who, according to the 60 - combing method, that is, in the order from the front left to the back left, and from the front right to the back right, comb their hair at a uniform speed 60 times (30 times on each side). Those whose hair loss count is more than 10 roots after combing and still more than 10 roots after a 2 - week washout period.

[0299] d. Those who have not had special hair styling treatments such as hair dyeing, perming, or styling within the past month.

[0300] e. Those who can understand the test process, voluntarily participate in the test, and sign a written informed consent form.

[0301] Those who meet the above conditions simultaneously are the subjects eligible for this test.

[0302] (2) Test equipment and materials

[0303] a. One NIKON D850 single - lens reflex camera, with about 45.75 million pixels.

[0304] b. Image shooting bracket: It can fix the position and direction of the subject's head and cooperate with the camera to take pictures of the top - of - the - head hair.

[0305] c. Youyingcai handheld dermoscope: Magnification from 50 to 1500 times, LED supplementary light, detection diameter 2.3 cm.

[0306] d. Comb: The tooth density is 7 teeth / cm, not replaced during the test process, and high - pressure sterilized after each use.

[0307] e. Hair analysis system software.

[0308] (3) Hair loss test evaluation method

[0309] During each follow - up visit of the subject, trained staff use the 60 - combing method to comb the subject's hair, count the number of fallen hairs and record it. The same - specification comb must be used throughout the test process.

[0310] (4) Local hair density evaluation method

[0311] A 1.5 cm × 1.5 cm hair - cutting area is fixed on the subject's head for positioning. Keep the hair - cutting area consistent during each follow - up visit. The hair is cut to a residual length not exceeding 1 mm; during the image acquisition process, the operator needs to place the subject in a comfortable position, place the dermoscope in the center of the hair - cutting area to take local scalp hair images. When taking pictures, the dermoscope lens is completely attached to the scalp and kept perpendicular, and check the clarity of the captured images. Load the pictures with image analysis software, count the number and density of hairs and record.

[0312] (5) Pull - test evaluation method

[0313] The hair pulling test is used to determine the tightness between the hair and the hair follicle. During each follow-up visit of the subject, a trained staff member slides a finger along the hair shaft of the subject while firmly pulling the hair away from the scalp, and the number of hairs extracted is counted.

[0314] (6) Basis for determining the effectiveness of the test items

[0315] a. At any visit time point during the trial, there is no significant increase in the hair loss count before and after using the test product, or the difference in hair loss count before and after (the hair loss count at a certain visit time point after product use - the hair loss count before product use) is significantly lower than that of the control group (p < 0.05).

[0316] b. Or, if there is no significant decrease in the local hair density, or the difference in hair density before and after (the hair density at a certain visit time point after product use - the hair density before product use) is significantly higher than that of the control group (p < 0.05), then the test product is considered to have anti - hair loss efficacy; otherwise, the test product is considered to have no anti - hair loss efficacy.

[0317] c. Or, at any visit time point during the trial, there is no significant increase in the hair pulling count before and after using the test product, or the difference in hair pulling count before and after (the hair pulling count at a certain visit time point after product use - the hair pulling count before product use) is significantly lower than that of the control group (p < 0.05).

[0318] (7) Relevance between the efficacy determination index and the efficacy claim

[0319] After using the product, the improvement in hair loss count, local hair density, and the decrease in the number of hairs in the hair pulling test all reflect that the product helps to improve or reduce hair loss and achieve the anti - hair loss effect.

[0320] (8) Results of the hair loss reduction test

[0321] After 30 Chinese adult female subjects continuously used the test product for 90 days according to the test method requirements, the hair loss count of the refreshing gel in Example 5 did not show a significant increase at each follow - up time point, and the overall hair density did not show a significant decrease at each follow - up time point; the difference in hair loss count of the refreshing gel in Example 5 was significantly lower than that of the control product group at the 8 - week follow - up after use (p < 0.05); the difference in overall hair density was higher than that of the formula in Comparative Example 3 and significantly higher than that of the control product group at the 12 - week follow - up after use (p < 0.05).

[0322] Based on the above test results, the refreshing gel in Example 5 has anti - hair loss efficacy.

[0323] (9) Results of the hair loss count

[0324] From Figure 6It can be seen that after the subjects used Example 5, the number of hair loss decreased by 54.4% at the 8th week compared to the starting point, and decreased by 73.3% at the 12th week. The number of hair loss in Comparative Example 3 also decreased at the 8th week and the 12th week compared to the starting point, which were 39.9% and 55.86% respectively, but the results were not as significant as those of Example 5. The number of hair loss in the negative control group (NC) increased at the 8th week and the 12th week, increasing from 32.43 at the starting point to 35 at the 12th week. This indicates that the formulation of Example 5 can more effectively reduce hair loss.

[0325] (10) Local hair density test results

[0326] According to Figure 7 the data analysis, through the statistical analysis of the average hair density rate in the test products measured, it shows that after the subjects used the refreshing gel of Example 5, the average hair density in the 1.5 cm × 1.5 cm area of the scalp increased from 148.35 per square centimeter at the starting point to 169.67 at the 12th week, and the density increased by 14.37%. For the subjects using Comparative Example 3, the hair density increased from 149.34 per square centimeter at the starting point to 163.32 at the 12th week, an increase of 9.36%, and the growth effect was not as significant as that of Example 5. The hair density of the negative control group (NC) decreased slightly within 12 weeks, from 147.23 per square centimeter to 142.87. These results indicate that Example 5 is more effective in promoting hair density.

[0327] (11) Hair pulling test results

[0328] According to Figure 8 the data, after using the refreshing gel of Example 5, the average number of hair pulled out by the subjects decreased from 4.35 at the starting point to 2.49 at the 12th week, a decrease of 42.8%, and a statistically significant change was observed at the 4th week. For the refreshing gel of Comparative Example 3, the number of hair pulled out also decreased from 4.03 at the starting point to 3.4 at the 12th week, a decrease of 15.6%, which was not as large as the decrease of Example 5. The number of hair pulled out in the negative control group (NC) did not change significantly during the entire study period, indicating that the refreshing gel of Example 5 is significantly effective in enhancing the tightness between hair and hair follicles.

[0329] 3. Analysis of self - assessment questionnaires before and after the subjects' use

[0330] During the product testing phase, no adverse events or serious adverse events were reported. However, hair and scalp conditions such as itching, dryness, and dandruff were data obtained as part of the "Subject Self-Assessment Questionnaire" during the Week 0 visit, and the problems reflected by these data were resolved after using the product of Example 5. During the study, no subjects had erythema, allergic reactions, folliculitis, oiliness, burning, or scalp sores, etc.

[0331] Subjects scored themselves on the self-assessment questionnaire before and after using Example 5.

[0332] The self-assessment is presented in the form of the average score of self-condition.

[0333] (i) Explanation of self-assessment data:

[0334] N = quantity, SD = standard deviation, M = month.

[0335] (ii) Explanation of the assessment score of hair loss rate:

[0336] 1: Feeling that more than 100 hairs fall out when combing hair every day (worst condition);

[0337] 2: Feeling that about 50 - 100 hairs fall out when combing hair every day;

[0338] 3: Feeling that about 30 - 50 hairs fall out when combing hair every day;

[0339] 4: Feeling that about 10 - 30 hairs fall out when combing hair every day;

[0340] 5: Feeling that less than 10 hairs fall out when combing hair every day (best condition).

[0341] (iii) Explanation of the assessment score of hair quality condition:

[0342] 1: Feeling that one's own hair is rough and frizzy (worst condition);

[0343] 2: Feeling that one's own hair is rough;

[0344] 3: Feeling that one's own hair is neither rough nor smooth;

[0345] 4: Feeling that one's own hair is relatively smooth;

[0346] 5: Feeling that one's own hair is very smooth (best condition).

[0347] (iv) Explanation of the assessment score of hair volume condition:

[0348] 1: Feeling that one's own hair volume is very small (worst condition - already affecting daily life);

[0349] 2: Feeling that one's own hair volume is relatively small;

[0350] 3: Feeling that the hair volume is normal;

[0351] 4: Feeling that the hair volume is relatively large;

[0352] 5: Feeling that the hair volume is very large (optimal condition).

[0353] (v) Instructions for scoring the dandruff assessment:

[0354] 1: Feeling that the dandruff is very much (the worst condition - has affected daily life);

[0355] 2: Feeling that the dandruff is relatively much;

[0356] 3: Feeling that the dandruff is normal;

[0357] 4: Feeling that the dandruff is less;

[0358] 5: Feeling that the dandruff is very little (optimal condition).

[0359] (vi) Instructions for scoring the scalp itching assessment:

[0360] 0: No itching sensation (optimal condition);

[0361] 1: Negligible itching sensation (has little impact);

[0362] 2: Slight itching sensation;

[0363] 3: Moderate itching sensation;

[0364] 4: Severe itching sensation;

[0365] 5: Very severe itching sensation (the worst condition - has affected daily life).

[0366] The evaluation results of the self-assessment questionnaire of the subjects before and after use are shown in Table 5 (the values in parentheses are the standard deviation values). It can be seen that with the increase of the continuous use time, the conditions of hair loss, hair quality, hair volume, dandruff, and scalp itching have significant improvements.

[0367] Table 5

[0368]

[0369] In summary, the composition of the present invention has excellent anti-hair loss effect and hair growth promoting effect, as well as significant DPPH free radical scavenging ability, which helps to reduce scalp inflammation, reduce dandruff and relieve scalp itching. And it also has excellent stability in the formula, so it is very suitable for scalp hair growth conditioning products (such as hair growth conditioning liquid, scalp essence, hair growth spray, hair growth hairspray, etc.), hair nourishing agents, drugs for promoting blood circulation, anti-hair loss agents, etc.

[0370] The embodiments described above merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation to the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the appended claims.

Claims

1. An application of a composition for preparing a preparation for promoting hair follicle cell growth, alleviating hair loss, promoting hair growth, and anti-oxidation; the composition comprises: Grape (VITIS VINIFERA) fruit extract, Scutellaria baicalensis root extract, Mourus Alba root extract, Saxifragia sarmentosa extract and Scutellaria alfalfa extract; among them, the grape fruit extract: Scutellaria baicalensis root extract: Mulberry root extract: Scutellaria sarmentosa extract: Scutellaria alfalfa extract are (0.004-0.05): (0.001-0.015): (0.001-0.015): (0.0002-0.004): (0.0015-0.02).

2. A composition for promoting the growth of hair follicle cells, alleviating hair loss, promoting hair growth and anti-oxidation, comprising: Grape fruit extract, scutellaria root extract, mulberry root extract, strawberry saxifrage extract and alpine scutellaria extract; among them, the grape fruit extract: scutellaria root extract: mulberry root extract: strawberry saxifrage extract: alpine scutellaria extract are (0.004-0.05):(0.001-0.015):(0.001-0.015):(0.0002-0.004):(0.0015-0.02).

3. A method for preparing a composition for promoting hair follicle cell growth, alleviating hair loss, promoting hair growth, and anti-oxidation, comprising: Grape fruit extract, scutellaria root extract, mulberry root extract, strawberry saxifrage extract and alpine scutellaria extract are mixed; wherein the ratio of grape fruit extract: scutellaria root extract: mulberry root extract: strawberry saxifrage extract: alpine scutellaria extract is (0.004-0.05):(0.001-0.015):(0.001-0.015):(0.0002-0.004):(0.0015-0.02).

4. The use according to claim 1, the composition according to claim 2 or the method according to claim 3, characterized in that: The grape fruit extract: scutellaria root extract: mulberry root extract: strawberry saxifrage extract: alpine scutellaria extract are (0.005-0.04):(0.0012-0.01):(0.0012-0.01):(0.0003-0.003):(0.002-0.015).

5. The use according to claim 1, the composition according to claim 2 or the method according to claim 3, characterized in that: The concentration of the total extract in the mixed system is 0.005-5%, preferably 0.005-3%, more preferably 0.005-2%; Preferably, the total extract is present in the mixed system in a diluted or concentrated state.

6. The use, composition or method according to claim 5, characterized in that The total extract is mixed in phase A, phase B and phase C, wherein phase A includes water, disodium EDTA, carbomer, sodium hydroxide, phenoxyethanol, ethyl ethyl glycerin; phase B includes essence and hydrogenated castor oil; phase C includes BLUE1; during mixing, the total extract is added to phase A, phase B and phase C to obtain a total mixture; or The total extract is mixed in phase A, phase B and phase C, wherein phase A includes water, grape fruit extract, disodium EDTA, sodium metabisulfite, sodium hydroxide, sodium sulfite, scutellaria root extract, butylene glycol, mulberry root extract, and strawberry saxifrage extract; phase B includes alpine scutellaria extract, glycerin, water, and citric acid; phase C includes dipotassium glycyrrhizate and water; during mixing, phase B is added to phase A, and then phase C is added to phases A and B to obtain a total mixture.

7. The use, composition or method according to claim 5, characterized in that: The grape fruit extract is obtained by extracting grape fruit with an organic solvent; preferably, the preparation method comprises: drying and crushing the grape fruit, soaking with an organic solvent, filtering to obtain a filtrate, and removing the organic solvent; optionally, the extract may be concentrated, purified, or diluted before use; The Scutellariae Radix extract is obtained by extracting Scutellariae Radix with water or a water-organic solvent mixture using Scutellariae Radix as a raw material; preferably, the preparation method comprises: drying and crushing the Scutellariae Radix, boiling it in water or a water-organic solvent mixture, filtering it after cooling and obtaining the filtrate, and concentrating it under reduced pressure, and optionally further comprising diluting it before use; The mulberry root extract is obtained by using mulberry roots as raw materials and extracting with organic solvents; preferably, the preparation method comprises: drying and crushing the mulberry roots, extracting with an organic solvent Soxhlet, and filtering to obtain a filtrate; optionally, the extract may also be concentrated, optionally, purified, and optionally, diluted before use; The strawberry saxifrage extract is obtained by water extraction using mulberry roots as raw materials; preferably, the preparation method comprises: drying and crushing the strawberry saxifrage, boiling in water, filtering after cooling and obtaining the filtrate, concentrating under reduced pressure, optionally further comprising purification, and optionally further comprising diluting before use; The alpine scutellaria extract is obtained by extracting the alpine scutellaria with a water-organic solvent mixture as the raw material; preferably, the preparation method includes: drying and crushing the alpine scutellaria; soaking it in water or a water-organic solvent mixture, or soaking it in methanol, or boiling it in a water-organic solvent mixture; filtering and obtaining the filtrate, concentrating it under reduced pressure and removing the solvent, and optionally also diluting it before use.

8. The use according to claim 1, the composition according to claim 2 or the method according to claim 3, characterized in that: The composition increases the expression of FGF-7, increases the expression of Wnt10B, increases the expression of β-catenin, decreases the expression of BMP4, and increases the DPPH free radical scavenging ability; or The composition also includes a pharmaceutically acceptable carrier.

9. A method for promoting the growth of hair follicle cells, alleviating hair loss, promoting hair growth, and anti-oxidation, comprising: Any of the above compositions is administered to a subject in need thereof.

10. A kit for promoting the growth of hair follicle cells, alleviating hair loss, promoting hair growth and anti-oxidation, comprising the composition according to claims 2, 4 to 7.

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