Traditional Chinese medicine composition for hair growth and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition with a hair growing function and a preparation method thereof. The external traditional Chinese medicine composition for hair growth comprises the following components in parts by weight: 10-30 parts of caulis spatholobi, 10-30 parts of macrocarpa nucuma, 10-30 parts of pinto beans, 2-15 parts of angelica sinensis, 5-15 parts of glossy privet fruits, 10-30 parts of radix polygoni multiflori preparata, 5-15 parts of cacumen biotae, 5-15 parts of mint and 5-15 parts of fresh ginger. By optimizing a product formula process and an extraction process, an optimal hair-growing product formula, extraction and other preparation processes are obtained, and the traditional Chinese medicine composition disclosed by the invention has the hair-growing effects of promoting hair growth, nourishing scalp and the like; the traditional Chinese medicine composition is not added with any pigment and preservative, the medicinal materials are all traditional Chinese medicines which can be taken orally and are non-toxic, and the traditional Chinese medicine composition can be used by old people, children and pregnant women, is non-irritant to skin and eyes and is safe to use.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an external Chinese medicine composition with hair growth function, an internal Chinese medicine composition and a preparation method thereof. Background Art

[0002] Chinese people generally have hair and scalp problems, among which hair loss is particularly serious. It is generally believed that hair loss is affected by many factors, including disease (endocrine disorders), psychology, drugs, genetics, spirit, age, environment and season. Western medicine believes that the main cause of hair loss is genetic factors and excessive secretion of male hormones. Among them, androgenic alopecia is seborrheic alopecia, which is the most common cause of hair loss; in addition, mental stress, work pressure, long-term anxiety, depression, staying up late and lack of sleep can also easily lead to hair loss. Traditional Chinese medicine believes that hair loss is related to internal dampness and heat, blood heat, liver depression and blood stasis, spleen deficiency and dampness, qi and blood deficiency, liver and kidney deficiency, etc.

[0003] Currently, the hair growth liquids on the market are divided into two categories: Western medicine and Chinese herbal medicine. Among them, Western medicine is mainly minoxidil and finasteride, but its side effects are large, causing patients to have side effects such as accelerated heartbeat and upper body pain, and it also has rebound effects. Chinese herbal hair growth liquids have problems such as poor hair growth effect and poor use experience. Summary of the invention

[0004] The object of the present invention is to provide an external hair growth Chinese medicinal composition.

[0005] Another object of the present invention is to provide a hair growth Chinese medicinal composition for oral administration.

[0006] Another object of the present invention is to provide a method for preparing the above-mentioned Chinese medicine composition.

[0007] According to a specific embodiment of the present invention, the external Chinese medicine composition for hair growth comprises the following components in parts by weight: 10-30 parts of Millettia spatholobi, 10-30 parts of Spatholobi nigra, 10-30 parts of Solanum lycopersicum, 2-15 parts of Angelica sinensis, 5-15 parts of Ligustrum lucidum, 10-30 parts of processed Polygonum multiflorum, 5-15 parts of Platycladus orientalis leaves, 5-15 parts of mint and 5-15 parts of ginger.

[0008] According to a specific embodiment of the present invention, the external Chinese medicine composition includes the following medicinal materials in parts by weight: 10 parts of Millettia reticulata, 10 parts of Spatholobi nigra, 10 parts of Solanum lycopodii, 2 parts of Angelica sinensis, 5 parts of Ligustrum lucidum, 10 parts of processed Polygonum multiflorum, 5 parts of Platycladus orientalis leaves, 5 parts of mint and 5 parts of ginger.

[0009] According to a specific embodiment of the present invention, the external Chinese medicine composition includes the following components by weight: 30 parts of Millettia reticulata, 30 parts of Spatholobi nigra, 30 parts of Solanum lycopersicum, 10 parts of Angelica sinensis, 15 parts of Ligustrum lucidum, 30 parts of processed Polygonum multiflorum, 15 parts of Platycladus orientalis leaves, 15 parts of mint and 15 parts of ginger.

[0010] According to a specific embodiment of the present invention, the external Chinese medicine composition includes the following components by weight: 20 parts of Millettia reticulata, 20 parts of Spatholobi nigra, 20 parts of Solanum lycopodii, 6 parts of Angelica sinensis, 10 parts of Ligustrum lucidum, 20 parts of processed Polygonum multiflorum, 10 parts of Platycladus orientalis leaves, 10 parts of mint and 10 parts of ginger.

[0011] According to a specific embodiment of the present invention, the oral Chinese medicine composition for hair growth includes the following components in parts by weight: 10-30 parts of Millettia spatholobi, 10-30 parts of Black Blood Caulis, 10-30 parts of Pig Kidney Bean, 2-15 parts of Angelica sinensis, 5-15 parts of Ligustrum lucidum, 10-30 parts of processed Polygonum multiflorum, 5-15 parts of mint, 5-15 parts of ginger, 10-30 parts of black beans, 10-30 parts of black sesame, and 10-30 parts of mulberries.

[0012] According to a specific embodiment of the present invention, the preparation method of the Chinese medicine composition comprises the following steps:

[0013] (1) taking each component and extracting it multiple times using a solvent;

[0014] First extraction: using ethanol-water solution (9:1, v / v) to extract, to obtain a first filter residue and a first extract;

[0015] Second extraction: extract the first filter residue with ethanol-water (5:5, v / v) to obtain a second filter residue and a second extract;

[0016] The third extraction: extracting the second filter residue with water to obtain a third filter residue and a third extract; combining the first extraction, the second extract and the third extract to obtain an extract;

[0017] (2) The extract obtained in step (1) is concentrated under reduced pressure to obtain an extract, which is the Chinese medicine composition.

[0018] Preferably, in step (1), the first extraction is performed at 35-55° C. for 6-72 h; and / or

[0019] The second extraction is carried out at 35-55°C for 6-72h.

[0020] Preferably, in step (1), during the third extraction, water is used for two extractions, and each extraction is performed for 0.5 h after boiling.

[0021] Preferably, in step (2), reduced pressure concentration is performed under the conditions of a vacuum degree of -0.08-0.1 MPa and a temperature of 0-55°C.

[0022] The efficacy of each medicine in the formula and modern research on its efficacy and pharmacology

[0023] Spatholobus suberectus is the dried vine stem of Spatholobus suberectus of the Leguminosae family. It has the effects of promoting blood circulation and replenishing blood, regulating menstruation and relieving pain, and relaxing meridians and activating collaterals; improving the levels of blood indicators related to iron deficiency anemia, and has anti-platelet aggregation, regulating lipid metabolism, protecting blood vessels, anti-tumor, anti-oxidation, anti-virus, regulating tyrosinase activity, and anti-depression effects; currently, the main compounds from Spatholobus suberectus are flavonoids (about half of the reported number), followed by phenolic acids, phenylpropanoids, terpenes and sterols; the flavonoid skeleton types include isoflavones, flavanols, chalcones, dihydroflavonoids and flavonoids; the active substances are mainly flavonoid compounds such as quercetin, (+)-catechin, formononetin, and luteolin.

[0024] Black Blood Twig is the root and stem of Mucuna macrocarpa, a plant of the genus Mucuna in the Leguminosae family. It has the effects of promoting blood circulation and nourishing blood, clearing the lungs and moistening dryness, and promoting blood circulation and activating collaterals. Modern pharmacology has proven that it has the effects of lowering blood lipids, improving blood rheology, and lowering blood sugar.

[0025] Pig kidney bean is the root and stem of Padbruggea filipes, a plant of the genus Padbruggea in the legume family. Its effects are similar to those of Millettia reticulata.

[0026] Angelica sinensis is the dried root of the Umbelliferae family, which has the effects of nourishing blood and activating blood circulation, regulating menstruation and relieving pain, and moistening the intestines and promoting bowel movements. It mainly contains compounds such as ligustilide, ferulic acid, and luteolin.

[0027] Processed Polygonum multiflorum is a processed product of the dried root tubers of Polygonum multiflorum, a plant of the Polygonaceae family. It has the effects of nourishing the liver and kidneys, replenishing the essence and blood, blackening the hair, and strengthening the tendons and bones. Its main chemical components are stilbene, flavonoids, and quinone compounds, followed by alkaloids, organic acids, sterols, lignans, etc. Modern pharmacological research shows that processed Polygonum multiflorum has anti-tumor, anti-inflammatory, immunomodulatory, anti-atherosclerotic, and antibacterial effects.

[0028] Fructus Ligustri Lucidi is the fruit of Ligustrum lucidum of the family Oleaceae. It has the effects of nourishing the liver and kidneys, strengthening the waist and knees, and improving eyesight. Its main chemical components are triterpenes, iridoids, flavonoids, and phenylethanol. Modern pharmacological research has shown that it has antioxidant, anti-inflammatory, antibacterial, anti-tumor, antiviral, anti-osteoporosis, hypoglycemic, lipid-regulating, immunomodulatory and liver-protecting effects.

[0029] Platycladus orientalis leaves are the young branches and leaves of Platycladus orientalis, a plant of the Cupressaceae family. They have the medicinal effects of cooling blood and stopping bleeding, resolving phlegm and relieving cough, and promoting hair growth and blackening hair. Their main components are volatile oils, flavonoids, tannins, etc.

[0030] Mint is the dried aerial part of the plant of the genus Menthae in the family Lamiaceae. It has the effects of dispersing wind-heat, clearing the head, relieving sore throat and relieving rashes, soothing the liver and promoting qi. Its main ingredients are L-menthol, luteolin-7-glucoside, amino acids, rosmarinic acid, and rheum officinale.

[0031] Ginger is the fresh rhizome of the ginger plant of the ginger family. It has the effects of dispelling cold, warming the middle and stopping vomiting, warming the lungs and stopping cough, and detoxifying. Modern pharmacological research has shown that it has anti-inflammatory, antioxidant, immunomodulatory, and neuroprotective effects.

[0032] Black sesame is the black seed of sesame of the genus Sesame of the Sesamum family. It has the effects of nourishing the liver and kidneys, nourishing the blood and improving the essence. Modern research shows that it is rich in nutrients such as vitamin B1 (thiamine), calcium, potassium, iron, zinc, and has the effects of improving immune function and anti-aging.

[0033] Black beans have the effects of promoting blood circulation, promoting diuresis, detoxifying, strengthening the spleen and kidney, and contain a variety of essential amino acids for the human body. Mulberries have the effects of detoxifying alcohol, promoting diuresis, reducing swelling, nourishing yin and replenishing blood, etc. Modern pharmacological research shows that they have the effects of improving immunity, improving sleep, delaying aging, lowering blood sugar, lowering blood lipids, and preventing atherosclerosis.

[0034] Beneficial effects of the present invention:

[0035] 1. Nourishing the scalp. The present invention uses Millettia reticulata and similar medicinal materials of Leguminosae that have the effects of promoting blood circulation and nourishing blood as the main medicine, and is supplemented by processed Polygonum multiflorum, ginger and other Chinese medicinal materials. It can deeply nourish the scalp and promote healthy hair growth.

[0036] 2. Promote the healthy growth of hair roots. The Chinese medicine composition of the present invention contains various ingredients such as flavonoids and volatile oils, which can improve blood circulation, reduce blood lipids, inhibit skin oil secretion, resist inflammation and oxidation, reduce hair follicle atrophy, prevent oxidative damage, inhibit oil secretion, and stimulate the healthy growth of hair roots. At the same time, the composition of the present invention can resist dandruff, relieve itching, and relieve scalp discomfort.

[0037] 3. By optimizing the product formula process and extraction process, the best preparation process such as the formula and extraction of the hair growth product for internal and external use is obtained. The product prepared by this preparation process has the effect of promoting hair growth and other hair growth effects. The Chinese medicine composition of the present invention does not add any pigments and preservatives, and the medicinal materials used are all oral, non-toxic Chinese medicines, which can be used by the elderly, children, and pregnant women, and are non-irritating to the skin and eyes and safe to use.

[0038] 4. The external Chinese medicine composition of the present invention is mainly aimed at the TCM pathogenesis of liver and kidney deficiency, blood stasis, and qi and blood deficiency of hair loss. The Millettia Spatholobi and Spatholobi nigra in the prescription are used as the main medicines, which have the effect of promoting blood circulation and unblocking collaterals, improving scalp blood circulation, and regulating hair follicle function; processed Polygonum multiflorum and Angelica sinensis are used as the ministerial medicines, which respectively play the effect of nourishing the liver and kidney and nourishing qi and blood, especially processed Polygonum multiflorum also has the effect of black hair, which helps to enhance the hair regeneration ability. The adjuvant ginger and Platycladus orientalis strengthen the local metabolic function by warming the meridians and dispersing cold, and promoting blood circulation, and improve the absorption efficiency of hair follicles to drugs. In addition, peppermint, as the adjuvant, has the effect of clearing away heat and detoxifying, dispersing wind and opening the orifices, which can not only promote the health of the scalp, but also make the drug better penetrate into the scalp.

[0039] 5. The oral Chinese medicine composition of the present invention regulates the fundamental cause of hair loss from the perspective of nourishing the liver and kidney, nourishing blood and activating blood circulation, and nourishing kidney essence. The main medicines are Millettia reticulata, Caulis nigra and Solanum lycopersicum, which mainly promote blood circulation and blood stasis, improve the circulation of qi and blood, and play a role in promoting blood circulation in the body and enhancing blood supply to hair follicles; the assistant medicines Polygonum multiflorum, Angelica sinensis and Fructus Ligustri Lucidi, respectively nourish the liver and kidney, nourish blood and activate blood circulation, and Polygonum multiflorum can also promote black hair regeneration and prevent premature alopecia. The adjuvant medicines include black beans, black sesame seeds and mulberries. These three medicines are classic medicines for nourishing kidney essence, have the effect of black hair, replenish human essence and blood, and help hair growth; the adjuvant medicines are mint and ginger, which harmonize the medicines and promote the effect of the medicines. Therefore, the oral prescription takes both internal and external factors into consideration by nourishing, activating blood circulation and regulating physical constitution, and effectively solves the problem of hair loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0041] Figure 1 The results of the negative control group (red line on the left) and the external application formula 3 group (red line on the right) on the hair growth of mice are shown (the 1st, 2nd and 3rd rows are 0, 3 and 14 days after administration, respectively).

[0042] Figure 2 The results of the effects of external application formula 2 (red line on the left) and external application formula 1 (red line on the right) on mouse hair growth are shown (the 1st, 2nd, and 3rd rows are 0, 3, and 14 days after administration, respectively).

[0043] Figure 3 The negative control group and external application formula 1-3 groups showed the effects of promoting the hair growth length of mice (unit: mm) (*P<0.05; **P<0.01; ***P<0.001).

[0044] Figure 4 Shows the efficacy results of extracts prepared by different extraction processes.

[0045] Figure 5 Showing the effect of extracts prepared by different extraction processes on promoting the growth length of mouse hair (unit: mm), (*P<0.05; **P<0.01; ***P<0.001).

[0046] Figure 6 Shown are the effects of the negative control group (left), the positive control group (middle), and the topical formulation 1 group (right) on hair growth in mice.

[0047] Figure 7 The results show the effects of the negative control group, the positive control group and the external application formula 1 group on the body surface temperature of mice (*P<0.05; **P<0.01; ***P<0.001).

[0048] Figure 8 The negative control group, positive control group and external application formula 1 group showed the effects of promoting the hair growth length of mice (unit: mm, (*P<0.05; **P<0.01; ***P<0.001).

[0049] Fig. 9 Shows the results of the effects of the positive control group (PCG) and external formula group 1 (EF1G) on skin inflammation in C56BL / 6 mice.

[0050] Fig.10 The results of the skin tissue structure effects of the negative control group (left), the positive control group (middle) and the topical formulation 1 group (right) are shown (HE, the first to third rows are 20X, and the fourth row is 40X).

[0051] Fig.11 The negative control group, positive control group and external application formula 1 group show the number of mouse hair follicles (left) and the thickness of the dermis (unit: mm, right figure).

[0052] Fig.12 Shows the effects of minoxidil and topical formula 1 on key proteins (immunofluorescence, Confocal detection, DAPI nuclear staining) in the Wnt / β-Catenin signaling pathway in the skin of mice with seborrheic alopecia (the left, middle, and right pictures are β-Catenin, β-Catenin, and GSK-3β in skin tissue, respectively).

[0053] Fig.13 Shows the effects of minoxidil and topical formula 1 on the expression of key proteins in the Wnt / β-Catenin signaling pathway in the skin of mice with seborrheic alopecia (Western Blot method,) (the left, middle and right pictures are β-Catenin, β-Catenin and GSK-3β in skin tissue, respectively).

[0054] Fig.14 Shows the effects of the negative control group and oral prescription groups 1-3 on hair growth in rats.

[0055] Fig.15 Shows the effects of the negative control group and oral prescription groups 1-3 on the hair growth length of rats (unit: mm), (*P<0.05; **P<0.01; ***P<0.001). DETAILED DESCRIPTION

[0056] To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0057] Example 1 Chinese medicine composition for external use

[0058] With 1 part by weight being 1 g, the Chinese medicine composition of the present invention comprises:

[0059] 1.1 The external Chinese medicine composition comprises the following components by weight: 10 parts of Millettia spatholobi, 10 parts of Spatholobi nigra, 10 parts of Solanum lycopersicum, 2 parts of Angelica sinensis, 5 parts of Ligustrum lucidum, 10 parts of processed Polygonum multiflorum, 5 parts of Platycladus orientalis leaves, 5 parts of mint and 5 parts of ginger.

[0060] 1.2 The external Chinese medicine composition includes the following components by weight: 30 parts of Millettia reticulata, 30 parts of Black Blood Caulis, 30 parts of Pig Kidney Bean, 10 parts of Angelica sinensis, 15 parts of Ligustrum lucidum, 30 parts of processed Polygonum multiflorum, 15 parts of Platycladus orientalis leaves, 15 parts of mint and 15 parts of ginger.

[0061] 1.3 The external Chinese medicine composition includes the following components by weight: 20 parts of Millettia reticulata, 20 parts of Black Blood Caulis, 20 parts of Pig Kidney Bean, 6 parts of Angelica sinensis, 10 parts of Ligustrum lucidum, 20 parts of processed Polygonum multiflorum, 10 parts of Platycladus orientalis leaves, 10 parts of mint and 10 parts of ginger.

[0062] The preparation method of the above-mentioned Chinese medicine composition is:

[0063] (1) taking each component and extracting it multiple times using a solvent;

[0064] First extraction: using ethanol-water solution (9:1, v / v) for extraction at a constant temperature of 50°C for 36 hours to obtain a first filter residue and a first extract;

[0065] Second extraction: The first filter residue was extracted with ethanol-water (5:5, v / v) at a constant temperature of 50°C for 36 h to obtain a second filter residue and a second extract;

[0066] The third extraction: extract the second filter residue with water, and extract for 0.5 hours after the water boils; extract twice with water to obtain the third filter residue and the third extract;

[0067] combining the first extraction, the second extraction and the third extraction to obtain an extraction solution;

[0068] (2) The extract obtained in step (1) is concentrated under reduced pressure at a vacuum degree of -0.1 MPa and a temperature of 40° C. to obtain an extract, which is a traditional Chinese medicine composition.

[0069] How to use: After shampooing, shake well and apply an appropriate amount on the head so that it fully contacts the scalp. Use your fingertips to gently press and rub the scalp for 5-10 minutes and then wash off.

[0070] Example 2 Effect verification of external Chinese medicine composition

[0071] 2.1 Effect of using the seborrheic alopecia mouse model

[0072] Six-week-old mice (strain: KM, male, weight: 40±10 g) were fed adaptively for 1 week, and then their back skin was depilated (depilatory range 2 cm×3 cm) using a depilatory cream to replicate the seborrheic alopecia mouse model.

[0073] The model group mice were randomly divided into negative control group (NCG), external formulation 1 group (EF1G), external formulation 2 group (EF2G) and external formulation 3 group (EF3G), with 10 mice in each group, and fed normal feed. At the same time, after one day of adaptation, the external formulation 1-3 groups were smeared with the corresponding formula, and the negative control group was smeared with aqueous solution, and the application was continued for 14 days. Within 14 days, photos of the back skin of the mice were taken.

[0074] External use formulation group 1: using the components of Example 1.1.

[0075] External use formula group 2: The components are the same as those of external use formula group 1, except that kidney beans are missing.

[0076] External use formula group 3: The components are the same as those of external use formula group 1, except that pig kidney bean and black blood vine are missing.

[0077] The effects of negative control group and external application formula 1-3 groups on hair growth in KM mice are shown in Figure 1-3 The results showed that the efficacy of external application formula group 1 was the best at 13 days.

[0078] 2.2 Effect of using the seborrheic alopecia mouse model:

[0079] Six-week-old mice (strain: C57BL / 6J, male, body weight: 40±10 g) were selected and, after adaptive feeding for 1 week, the seborrheic alopecia mouse model was established according to the above method.

[0080] The model mice were randomly divided into a water extract group, a 50% ethanol extract group, and a 90% ethanol extract group, with 10 mice in each group, and fed with normal feed. After one day of adaptation, the drug was applied continuously for 14 days. Within 14 days, photos of the back skin of the mice were taken.

[0081] 90% ethanol gradient extract group: the preparation method is the same as Example 1.

[0082] Water extract group: The first extraction and the second extraction steps were omitted, and only water was used for two extractions, each time after boiling for 0.5 hours. The extracts were combined to obtain a water extract group; the other steps were the same as in Example 1.

[0083] 50% ethanol gradient extract group: the first extraction step was omitted, and ethanol-water (5:5; v / v) and water were used for extraction in sequence, and the other steps were the same as in Example 1.

[0084] The results are as follows Figure 4 and Figure 5 As shown, the effect of 90% ethanol gradient extract is better than that of 50% ethanol gradient extract, and the effect of 50% ethanol gradient extract is better than that of water extract.

[0085] 2.3 Comparison of the effect with minoxidil

[0086] Seborrheic alopecia model mice were randomly divided into negative control group (NCG), positive control group (5% minoxidil, PCG) and optimal external formula and optimal extraction process group (external formula group 1, EF1G), with 10 mice in each group, and they were fed normal feed.

[0087] At the same time, the 2cm×3cm area of ​​the mouse back skin was depilated, and after one day of adaptation, the drug was applied continuously for 14 days. Photos of the mouse back skin were taken and the mice were killed 14 days later. The mice were fasted but not watered for 12 hours and anesthetized by intraperitoneal injection of 20% urethane (dose: 7.5ml / kg). The skin of the drug-applied area was taken for sectioning and mechanism research. Five hairs were taken from each mouse to measure the hair length and calculate the average value.

[0088] Hematoxylin-eosin (HE) staining: The back skin of 10 mice in each group was fixed with 4% paraformaldehyde fixative, and the back skin tissue of the mice was embedded in paraffin, 5 μm sections were dewaxed, and HE staining was performed, and the skin sections were observed and photographed under a microscope. The differentiation degree of the skin hair follicle sections was observed by scanning the HE-stained skin sections. The hair follicle differentiation changed from spherical to flagellar, and the degree of differentiation increased.

[0089] Use Image J software to mark and count the HE-stained hair follicles. Open the image in ImageJ → Type 8-bit, Process → Enhance Contrast, and the Enhance Contrast window will pop up. Fill 0.3 in Saturated pixels, check Normalize, and uncheck Equalize histogram. Process → Find Maxima, and the Find Maxima window will pop up. Select Count for output type, and you can see that all the hair follicles in the image are marked. Compare the average number of hair follicles between each group.

[0090] Use Image J software to measure the HE-stained sections, select the image, set the known length, Analyze-SetMeasurements, click Measure, and randomly select three locations on each section to measure the dermis thickness and take the average value.

[0091] Western blotting: Four mice were randomly selected from each group, and the dorsal skin was taken. The total protein was extracted by lysis buffer and the concentration was determined by BCA method. Equal amount of protein was subjected to 5% SDS-PAGE electrophoresis, transferred to the membrane, blocked with fast blocking solution at room temperature for 15 min, and 50 μg protein samples were analyzed by Western blotting with corresponding antibodies; Wnt-3α (1:1000, Santa), Akt (1:1000, Santa), LRP5 (1:1000, Santa), β-actin (1:2000, SantaCruz), VEGF (1:1000, Santa), β-Catenin (1:2000, Santa), frizzled-1 (1:1000, Santa), Akt (1:1000, Santa). Western blot was analyzed by chemiluminescence detector iBrightFL1000 (Invitrogen, Waltham, MA, USA).

[0092] Immunofluorescence: Dewax the skin paraffin sections in a 60°C oven for 30 minutes, and place the sections in xylene for 20 minutes, twice. Then place them in 100%, 100%, 95%, and 80% ethanol for 5 minutes each, and then rinse them in distilled water 3 times, 3 minutes each time. Add 1×EDTA (pH8.0) antigen repair solution enough to submerge the sections in a 1.5L pressure cooker and heat to boiling. Place the dewaxed sections on the staining rack and set the pressure to maintain for 4 minutes. Take out the sections after the solution temperature is balanced with room temperature. 3% HO, incubate at room temperature for 10 minutes, 5% (goat) blocking serum, room temperature for 1 hour. Incubate with blind primary antibody, fluorescent secondary antibody (1:2000, diluted with 5% BSA), and quencher DAPI for color development. Images were taken using a laser scanning confocal microscope (FV300).

[0093] The results of the effects on hair growth, body surface temperature and hair length in the negative control group (NCG), the positive control group (minoxidil group, PCG) and the best external formula group (external formula 1, EF1G) in mice with seborrheic alopecia are shown in Figure 6-8 .

[0094] The results showed that both topical formula 1 and minoxidil could promote hair growth in mice, and the efficacy of topical formula 1 was better than that of minoxidil.

[0095] Body surface temperature is related to hair growth.

[0096] The results of the effect of external application formula 1 on inflammation of mouse skin are shown in Fig. 9 As shown, the results showed that the degree of inflammation caused by Formula 1 was much lower than that of minoxidil.

[0097] The results of the effects on skin tissue structure, hair follicle number and dermis thickness in the negative control group, minoxidil group and optimal topical formula group are shown in Fig.10 and Fig.11 The results showed that both topical formula 1 and minoxidil could promote the differentiation of small hair follicles and nourish the dermis, and the efficacy of topical formula 1 was better than that of minoxidil.

[0098] The results of minoxidil and topical formula 1 on the expression of key proteins in the Wnt / β-Catenin signaling pathway in the skin of mice with seborrheic alopecia are shown in Fig.12 and Fig.13 .

[0099] The results of immunofluorescence detection showed that compared with the negative control group, the fluorescence intensity of β-Catenin in the external formula 1 group was significantly enhanced, and the fluorescence intensity of GSK-3β was significantly weakened, while the positive drug group had little change. The results of Western Blot showed that compared with the negative control group, the relative contents of Wnt-3α, LRP5, GSK-3β, β-Catenin, AKT, VEGF, and frizzled-1 proteins in the external formula 1 group were significantly increased, while the positive drug group had little change, indicating that the pharmacodynamic mechanism of formula 1 is related to the key proteins Wnt-3α, LRP5, GSK-3β, β-Catenin, AKT, VEGF, and frizzled-1 that affect the Wnt / β-Catenin pathway.

[0100] The above results showed that the three topical formulas could promote the growth of mice with seborrheic alopecia, and the efficacy of topical formula 1 was the best; the best extraction process was ethanol-water (9 1, 5 5, 0 10; v / v) gradient extraction. The efficacy of topical formula 1 was better than that of the positive drug minoxidil, and its mechanism was related to the effects on key proteins of the Wnt / β-Catenin pathway, Wnt-3α, LRP5, GSK-3β, β-Catenin, AKT, VEGF, and frizzled-1.

[0101] Example 3 Oral Chinese medicine composition

[0102] With 1 part by weight being 1 g, the Chinese medicine composition of the present invention comprises:

[0103] 3.1 The Chinese medicine composition for oral administration comprises the following components in parts by weight: 10 parts of Millettia reticulata, 10 parts of Black Blood Caulis, 10 parts of Pig Kidney Bean, 5 parts of Angelica sinensis, 5 parts of Ligustrum lucidum, 10 parts of processed Polygonum multiflorum, 5 parts of mint, 5 parts of ginger, 1 part of black bean, 1 part of black sesame, and 1 part of mulberry.

[0104] 3.2 The Chinese medicine composition for oral administration comprises the following components in parts by weight: 30 parts of Millettia reticulata, 30 parts of Black Blood Caulis, 30 parts of Pig Kidney Bean, 10 parts of Angelica sinensis, 15 parts of Ligustrum lucidum, 30 parts of processed Polygonum multiflorum, 15 parts of mint, 15 parts of ginger, 30 parts of black beans, 30 parts of black sesame, and 30 parts of mulberries.

[0105] 3.3 The Chinese medicine composition for oral administration includes the following components by weight: 20 parts of Millettia reticulata, 20 parts of Black Blood Caulis, 20 parts of Pig Kidney Bean, 6 parts of Angelica sinensis, 10 parts of Ligustrum lucidum, 20 parts of processed Polygonum multiflorum, 10 parts of mint, 10 parts of ginger, 15 parts of black beans, 15 parts of black sesame, and 15 parts of mulberries.

[0106] The preparation method of the above-mentioned Chinese medicine composition is the same as the preparation method in Example 1.

[0107] Directions for use: Put 2 ml of extract in a cup, then add 200 ml of warm water, stir evenly with a spoon and take orally, once in the morning and once in the evening.

[0108] Example 4 Oral Formula Pharmacodynamics Study

[0109] Forty rats aged 77-83 days (species: SD, male, weight: 350±30g) were fed adaptively for 1 week, and then their back skin was depilated (depilatory range 5cm×8cm) using depilatory cream to replicate the seborrheic alopecia rat model.

[0110] The model group rats were randomly divided into negative control group (NCG), oral prescription group 1 (OP1G), oral prescription group 2 (OP2G) and oral prescription group 3 (OP3G), with 10 rats in each group, and fed with normal feed. At the same time, after one day of adaptation, the drug was applied continuously for 18 days. Within 18 days, photos of the back skin of the rats were taken.

[0111] Oral formulation 3: components of Example 3.1.

[0112] Oral formula 2: The ingredients are the same as those of oral formula 1, except that kidney beans are missing.

[0113] Oral formula 1: The ingredients are the same as those of oral formula 1, except that pig kidney beans and black blood vine are missing.

[0114] The results of the effects of each group on rat hair growth are shown in Fig.14 The results showed that each group could promote the hair growth of rats, and the oral prescriptions 1-3 could promote the hair growth of rats. The efficacy of the three prescriptions was slightly different, and the efficacy of oral prescription 3 was the best.

[0115] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art who is familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A Chinese medicine composition for external use for hair growth, characterized in that: The Chinese medicine composition comprises the following components in parts by weight: 10-30 parts of Millettia reticulata, 10-30 parts of Spatholobi nigra, 10-30 parts of Solanum lycopodii, 2-15 parts of Angelica sinensis, 5-15 parts of Ligustrum lucidum, 10-30 parts of processed Polygonum multiflorum, 5-15 parts of Platycladus orientalis leaves, 5-15 parts of mint and 5-15 parts of ginger.

2. The external Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition comprises the following components by weight: 10 parts of Millettia reticulata, 10 parts of Spatholobi nigra, 10 parts of Solanum lycopodii, 2 parts of Angelica sinensis, 5 parts of Ligustrum lucidum, 10 parts of processed Polygonum multiflorum, 5 parts of Platycladus orientalis leaves, 5 parts of mint and 5 parts of ginger.

3. The external Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition comprises the following components by weight: 30 parts of Millettia reticulata, 30 parts of Spatholobi nigra, 30 parts of Solanum lycopersicum, 10 parts of Angelica sinensis, 15 parts of Ligustrum lucidum, 30 parts of processed Polygonum multiflorum, 15 parts of Platycladus orientalis leaves, 15 parts of mint and 15 parts of ginger.

4. The external Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition comprises the following components by weight: 20 parts of Millettia reticulata, 20 parts of Spatholobi nigra, 20 parts of Solanum lycopersicum, 6 parts of Angelica sinensis, 10 parts of Ligustrum lucidum, 20 parts of processed Polygonum multiflorum, 10 parts of Platycladus orientalis leaves, 10 parts of mint and 10 parts of ginger.

5. The method for preparing the Chinese medicine composition according to any one of claims 1 to 4, characterized in that: The preparation method comprises the following steps: (1) taking each component and extracting it multiple times using a solvent; First extraction: using ethanol-water solution (9:1, v / v) to extract, to obtain a first filter residue and a first extract; Second extraction: extract the first filter residue with ethanol-water (5:5, v / v) to obtain a second filter residue and a second extract; The third extraction: extracting the second filter residue with water to obtain a third filter residue and a third extract; combining the first extraction, the second extract and the third extract to obtain an extract; (2) The extract obtained in step (1) is concentrated under reduced pressure to obtain an extract, which is the Chinese medicine composition.

6. The preparation method according to claim 5, characterized in that: In step (1), the first extraction is performed at 35-55° C. for 6-72 hours; and / or, The second extraction is carried out at 35-55°C for 6-72h.

7. The preparation method according to claim 5, characterized in that: In step (2), reduced pressure concentration is performed under the conditions of vacuum degree of -0.08-0.1 MPa and temperature of 0-55°C.

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