Traditional Chinese medicine composition for treating alopecia, traditional Chinese medicine preparation and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating alopecia, a traditional Chinese medicine preparation and application thereof. The traditional Chinese medicine composition comprises the following traditional Chinese medicine raw materials in parts by weight: 1-50 parts of radix polygoni multiflori preparata, 1-50 parts of fructus ligustri lucidi, 1-50 parts of herba ecliptae, 1-50 parts of rhizoma polygonati, 1-50 parts of mulberries, 1-50 parts of radix angelicae sinensis, 5-100 parts of radix salviae miltiorrhizae, 1-50 parts of rhizoma chuanxiong and 1-50 parts of flos carthami. The traditional Chinese medicine composition can inhibit apoptosis, prolong the hair growth cycle and promote hair growth.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine, and in particular to a traditional Chinese medicine composition and preparation for treating hair loss and applications thereof. Background Art

[0002] Androgenetic alopecia (AGA) is a terminal hair loss that develops gradually after puberty and is classified as either male pattern hair loss (MPHL) or female pattern hair loss (FPHL). In Traditional Chinese Medicine (TCM), AGA is often referred to as "hair worm hair loss," "hair worm ringworm," or "white dandruff." It is characterized by hair loss and miniaturization of hair follicles. The condition is most common after puberty, with incidence increasing with age, affecting approximately half of the world's population.

[0003] Currently, commonly used AGA treatments include topical minoxidil and oral finasteride, but their side effects, particularly impacting male sexual function, have been widely reported. In contrast, Traditional Chinese Medicine (TCM) offers extensive experience in treating AGA, demonstrating significant efficacy, minimal side effects, and is economical and practical. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a traditional Chinese medicine composition, a traditional Chinese medicine preparation and applications thereof for treating hair loss.

[0005] A first aspect of the present invention provides a traditional Chinese medicine composition for treating hair loss, wherein the raw materials are mixed by weight as follows: 1-50 parts of processed Polygonum multiflorum, 1-50 parts of Ligustrum lucidum fruit, 1-50 parts of Eclipta prostrata, 1-50 parts of Polygonatum sibiricum, 1-50 parts of mulberry, 1-50 parts of Angelica sinensis, 5-100 parts of Salvia miltiorrhiza, 1-50 parts of Chuanxiong and 1-50 parts of Carthamus tinctorius.

[0006] In the prior art, it is generally believed that hair loss is related to internal organs such as the liver, spleen, and kidneys, and the pathogenesis is mostly manifested as deficiency of the root and excess of the superficial symptoms. In the early stage, excess symptoms such as damp heat and liver depression are the main ones, while in the later stage, deficiency symptoms such as liver and kidney deficiency and insufficient qi and blood are the main ones. The Chinese medicine composition of the present invention is a summary of the inventor's many years of clinical experience, and advocates the treatment of hair loss by nourishing the liver and kidneys, replenishing qi and activating blood circulation. Studies have shown that the Chinese medicine composition can increase the expression level of key proteins in the Wnt signaling pathway, reduce androgen content, inhibit the binding of androgens to AR, thereby reducing the degradation of β-catenin, and then regulating the Wnt / β-catenin signaling pathway. In addition, the Chinese medicine composition can also inhibit cell apoptosis, prolong the hair growth cycle, and promote hair growth.

[0007] Processed Shouwu is a processed product of Polygonum multiflorum, which has the effect of nourishing and strengthening the kidneys.

[0008] In some embodiments, the weight ratio of the Chinese medicinal materials is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts or any value therebetween.

[0009] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts or any value therebetween.

[0010] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts or any value therebetween.

[0011] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts or any value therebetween.

[0012] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts of mulberry or any value therebetween.

[0013] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts of angelica or any value therebetween.

[0014] In some embodiments, the weight ratio of the Chinese medicinal raw materials is: 5 parts, 7 parts, 10 parts, 12 parts, 15 parts, 17 parts, 19 parts, 21 parts, 23 parts, 25 parts, 27 parts, 29 parts, 30 parts, 32 parts, 34 parts, 36 parts, 40 parts, 60 parts, 80 parts, 100 parts or any value therebetween.

[0015] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts of Chuanxiong or any value therebetween.

[0016] In some embodiments, the weight ratio of the Chinese medicinal material is: 1 part, 3 parts, 5 parts, 7 parts, 10 parts, 12 parts, 14 parts, 16 parts, 18 parts, 20 parts, 22 parts, 24 parts, 26 parts, 30 parts, 35 parts, 40 parts, 45 parts, 50 parts of safflower or any value therebetween.

[0017] In some embodiments, the weight ratio of the Chinese medicinal raw materials is: 5-20 parts of processed Polygonum multiflorum, 5-20 parts of Ligustrum lucidum fruit, 5-20 parts of Eclipta prostrata, 5-20 parts of Polygonatum sibiricum, 5-20 parts of mulberry, 5-20 parts of Angelica sinensis, 10-30 parts of Salvia miltiorrhiza, 5-20 parts of Chuanxiong and 5-20 parts of Carthamus tinctorius.

[0018] In some embodiments, the Chinese medicinal raw materials are proportioned by weight as follows: 8-12 parts of processed Polygonum multiflorum, 8-12 parts of Ligustrum lucidum fruit, 8-12 parts of Eclipta prostrata, 8-12 parts of Polygonatum sibiricum, 8-12 parts of mulberry, 8-12 parts of Angelica sinensis, 12-17 parts of Salvia miltiorrhiza, 8-12 parts of Chuanxiong and 8-12 parts of Carthamus tinctorius.

[0019] In some embodiments, the weight ratio of the Chinese medicinal raw materials is: 10 parts of processed Polygonum multiflorum, 10 parts of Ligustrum lucidum fruit, 10 parts of Eclipta prostrata, 10 parts of Polygonatum sibiricum, 10 parts of mulberry, 10 parts of Angelica sinensis, 15 parts of Salvia miltiorrhiza, 10 parts of Chuanxiong and 10 parts of Carthamus tinctorius.

[0020] The second aspect of the present invention provides a traditional Chinese medicine preparation, comprising the traditional Chinese medicine composition described in the first aspect.

[0021] In some embodiments, the dosage form of the traditional Chinese medicine preparation is selected from one or more of granules, decoctions, tablets, lotions, ointments, creams, hydrogels, or essential oils.

[0022] In some embodiments, the preparation is in the form of granules and / or decoction.

[0023] In some embodiments, when the dosage form of the traditional Chinese medicine preparation is granules, the granules further include excipients.

[0024] In some embodiments, the excipients include inulin, sucralose, and isomalt.

[0025] In some embodiments, when the dosage form of the traditional Chinese medicine preparation is granules, the preparation method of the granules comprises:

[0026] (1) Pretreatment: Grind isomalt and pass through an 80-mesh sieve for later use. If the remaining materials are lumpy, pass through an 80-mesh sieve;

[0027] (2) Decoction: According to the ingredient list, weigh processed Polygonum multiflorum, Ligustrum lucidum fruit, Eclipta prostrata, Polygonatum sibiricum, mulberry, Chinese angelica, Salvia miltiorrhiza, Ligusticum chuanxiong, and Carthamus tinctorius, add 10 times the amount of water and decoct twice, the first decoction for 2 hours, the second decoction for 1 hour, filter, and set aside;

[0028] (3) Concentration: The filtrate is concentrated under reduced pressure (-0.06 to -0.10 MPa, ≤70°C) to a thick paste with a relative density of 1.15 to 1.25 (60 ± 2°C);

[0029] (4) Drying: Dry the thick paste under reduced pressure (-0.06 to -0.10 MPa, ≤70°C) until dry, grind through an 80-mesh sieve to obtain dry extract powder, and set aside;

[0030] (5) Granulation: Weigh isomalt, inulin, and dry extract powder according to the formula amount, place them together in a high-efficiency wet granulator and mix for 5 minutes to obtain a mixed powder; dissolve sucralose in 95% ethanol (about 18% of the total formula amount; the ethanol concentration and amount can be adjusted according to actual conditions) as a wetting agent to prepare a soft material, granulate into 14 mesh, dry at 55±5°C (the moisture content of the granules is controlled to be below 5%), and granulate into 14 mesh to obtain dry granules for later use;

[0031] (6) Inner packaging: composite film packaging, 10g / bag;

[0032] (7) Outer packaging: labeling, boxing, and packing, and storage after passing inspection;

[0033] (8) Inspection and warehousing: Finished products are put into storage after passing the inspection.

[0034] In some embodiments, the isomalt is added to a total amount (small test yield of about 20-25%).

[0035] In some embodiments, the amount of inulin is 3 to 10 parts, for example, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts or any value therebetween.

[0036] In some embodiments, the amount of sucralose is 0.04 to 0.16 parts, for example, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.16 parts, or any value therebetween.

[0037] In some embodiments, when the dosage form of the traditional Chinese medicine preparation is a decoction, the preparation method of the decoction comprises:

[0038] (1) Weighing the raw materials: Weigh the raw materials of the composition according to the formula: Polygonum multiflorum, Ligustrum lucidum fruit, Eclipta prostrata, Polygonatum sibiricum, mulberry, Angelica sinensis, Salvia miltiorrhiza, Ligusticum chuanxiong, and Carthamus tinctorius;

[0039] (2) Soaking: Put the formulated amount of raw materials into a container dedicated to Chinese medicine decoction, add 6 to 8 times the amount of cold water (drinking water that meets national health standards), and soak for 30 minutes to prepare for decoction;

[0040] (3) First decoction: Boil the soaked Chinese medicine and water over high heat until boiling, then simmer for 10-20 minutes over low heat. Turn off the heat, pour out the hot liquid and filter it to obtain 200 mL of liquid. Store in a sterilized container for later use.

[0041] (4) Second decoction: Add 6 to 8 times the amount of cold water (drinking water that meets national health standards) to the residue from the first decoction, boil over high heat until boiling, then switch to low heat and continue boiling for 10 to 20 minutes. Turn off the heat, pour out the hot liquid and filter it, and squeeze the residue to filter it, obtaining a total of 200 mL of liquid.

[0042] (5) Mixing: Pour the second decoction into the container containing the first decoction and mix evenly to obtain the decoction. Then, package the decoction in 200 mL bags.

[0043] The third aspect of the present invention provides a use of the traditional Chinese medicine composition described in the first aspect or the traditional Chinese medicine preparation described in the second aspect in the preparation of a medicament for treating or preventing hair loss.

[0044] In some embodiments, the hair loss is androgenetic alopecia.

[0045] The fourth aspect of the present invention provides a use of the traditional Chinese medicine composition of the first aspect or the traditional Chinese medicine preparation of the second aspect in a product for promoting hair growth.

[0046] The fifth aspect of the present invention provides a use of the traditional Chinese medicine composition described in the first aspect or the traditional Chinese medicine preparation described in the second aspect in the preparation of a drug for lowering androgen levels, inhibiting 5-AR, promoting Wnt / β-catenin signaling pathway activity and / or inhibiting cell apoptosis.

[0047] Compared with the prior art, the present invention has the following beneficial effects:

[0048] Androgenic alopecia, also known as seborrheic alopecia, is a multi-gene inherited disorder whose pathogenesis is linked to androgen metabolism, characterized by elevated levels of dihydrotestosterone (DHT) in the patient's serum and scalp. Currently, Western medical treatments (such as finasteride and minoxidil) and surgical therapies (autologous hair follicle transplantation) are ineffective in effectively treating hair loss and are associated with varying degrees of adverse reactions and recurrence after discontinuation. Traditional Chinese medicine (TCM) offers unique therapeutic benefits, holistic regulation, and minimal adverse reactions in the prevention and treatment of hair loss.

[0049] Traditional Chinese Medicine refers to this condition as "worm-eaten hair loss" or "hair tinea." The Suwen (Suwen) Treatise on the Original Nature of Ancient Things states: "At the age of seven, a woman's kidney qi is strong, her teeth grow and her hair grows... At the age of fifty-seven, the Yangming meridian weakens, her face begins to become sallow, and her hair begins to fall..." "At the age of eight, a man's kidney qi is strong, his hair grows and his teeth grow... At the age of fifty-eight, his kidney qi weakens, his hair falls, and his teeth become dry." This demonstrates the close correlation between hair growth and the strength and weakness of kidney qi. The Yilin Gaicuo (Yilin Gaicuo) states: "Blood stasis within the skin and flesh blocks the blood circulation, preventing new blood from nourishing the hair, resulting in hair loss and failure to grow. Hair loss without disease is also due to blood stasis." This suggests that blood stasis is also a cause of this condition. The Yizong Jinjian (Yizong Jinjian) states: "Although hair belongs to the five internal organs, it is all produced by blood, and therefore desires lustre." This suggests a close connection between the five internal organs, qi, and blood, and hair growth. The Huangdi Neijing (Yellow Emperor's Internal Classic) states: "Hair is the residue of blood, and the kidney's essence is reflected in the hair." As can be seen from this, liver and kidney deficiency and qi stagnation and blood stasis are the main pathogenesis of this disease. Liver and kidney deficiency cannot transform essence and blood, resulting in an empty hair root and a lack of growth source; qi stagnation and blood stasis prevent blood from nourishing the hair, causing large scalp hair to fall out. To this end, the present invention uses Polygonum multiflorum as the main drug to nourish the liver and kidney, benefit essence and blood, and blacken hair. The Compendium of Materia Medica states that Polygonum multiflorum "can nourish blood and benefit the liver, consolidate essence and benefit the kidney, strengthen tendons and bones, and blacken hair, making it a good tonic." Assistant herbs: Ligustrum lucidum fruit nourishes the liver and kidneys and blackens hair. "Compendium of Materia Medica" states that Ligustrum lucidum fruit "benefits the liver and kidneys, soothes the five internal organs, strengthens the waist and knees, brightens the eyes and ears, blackens the hair, replenishes wind deficiency, and cures all ailments." Herba Ecliptae (Herba Ecliptae) nourishes the liver and kidneys. "Compendium of Materia Medica" states that "Herba Ecliptae enters the kidneys to replenish yin and promote hair growth. It also enters the blood, cooling the blood and stopping bleeding." Polygonatum sibiricum nourishes qi and yin, strengthens the spleen, moistens the lungs, and benefits the kidneys. Mulberry fruit nourishes yin and blood, promotes fluid production and moistens dryness. "Compendium of Materia Medica in Southern Yunnan" states that "Mulberry fruit benefits the kidneys and strengthens essence; long-term use leads to black hair and bright eyesight." Together, these herbs nourish the liver and kidneys, nourish blood and blacken hair. Auxiliary herbs: Angelica sinensis nourishes blood and invigorates blood circulation; Salvia miltiorrhiza invigorates blood circulation and removes blood stasis; Ligusticum chuanxiong invigorates blood circulation and qi; and Safflower nourishes blood, invigorates blood circulation and removes blood stasis. Together, these herbs nourish the liver and kidneys, invigorate blood circulation and remove blood stasis, and promote hair growth and blacken hair. It is suitable for patients with liver and kidney deficiency and qi stagnation and blood stasis-induced hair loss.

[0050] Compared to the Chinese patent CN 103239634 B, the present invention's traditional Chinese medicine formula consists of raw rehmannia root, cooked rehmannia root, angelica root, ligusticum chuanxiong, white peony root, glossy privet fruit, wolfberry fruit, dodder seed, mulberry, black sesame, atractylodes macrocephala, poria cocos, eclipta prostrata, oriental arborvitae, salvia miltiorrhiza, millet vine, processed shouwu root, polygonatum sibiricum, cordyceps sinensis, gastrodia elata, calamus tatarinowii, and dittany bark. The present invention's formula is refined, and according to the Yellow Emperor's Classic of Internal Medicine, "hair is the residue of blood, and the kidney's essence is in the hair," the addition of safflower enhances blood-nourishing, blood-activating, and blood-stasis-removing properties.

[0051] Compared to Chinese patent number CN 106822399 A, the present invention's traditional Chinese medicine formula consists of angelica sinensis, polygonum multiflorum, ligusticum chuanxiong, rehmannia root, dodder seed, codonopsis pilosula, eclipta prostrata, eclipta herba, wolfberry, glossy privet fruit, white peony root, moutan bark, oriental water chestnut, and safflower. The present invention's formula is refined, with the addition of salvia miltiorrhiza to promote blood circulation and dispel blood stasis; polygonatum to tonify qi and nourish yin, strengthen the spleen, moisten the lungs, and benefit the kidneys; and mulberry to nourish yin and blood, resulting in superior liver and kidney nourishment, blood-nourishing, blood-activating, and blood-stasis-removing effects.

[0052] Compared with Chinese patent number CN 107362235 A, the traditional Chinese medicine formula of CN 107362235 A comprises processed Polygonum multiflorum, moutan bark, rehmannia root, glossy privet fruit, angelica root, ligusticum chuanxiong, raw rehmannia root, red peony root, slough of snake skin, American ginseng, mulberry, donkey-hide gelatin, black sesame, and Eclipta prostrata. The present invention has a refined formula, with the addition of polygonatum to tonify qi and nourish yin, strengthen the spleen, moisten the lungs, and benefit the kidneys; safflower to nourish blood, promote blood circulation, and remove blood stasis; and salvia miltiorrhiza to promote blood circulation and remove blood stasis, resulting in superior liver and kidney nourishing, blood-nourishing, blood-activating, and blood-stasis-removing effects.

[0053] Compared with Chinese patent number CN101485831A, the present invention comprises the following internal medicines: Codonopsis pilosula, Astragalus membranaceus, Angelica sinensis, Radix Polygoni Multiflori Preparata, Platycladus orientalis leaves, Eclipta prostrata, Rehmannia root, Morus alba, Ligustrum lucidum fruit, Candida altissima leaves, Polygonatum sibiricum, Lycium barbarum fruit, Black sesame, and Cuscuta seed; while the external medicines comprise Polygonum multiflorum, Salvia miltiorrhiza, Platycladus orientalis leaves, Candida altissima leaves, Ligustrum lucidum fruit, Ligustrum lucidum rhizome, Dictamnus cortex, Eclipta prostrata, Cuscuta seed, Allium sibiricum seeds, and ginger. The present invention has a refined formula, with the addition of safflower to nourish blood, promote blood circulation, and dispel blood stasis; Salvia miltiorrhiza to promote blood circulation, and Ligusticum chuanxiong to promote blood circulation and qi circulation, resulting in superior blood-nourishing, blood-activating, and blood-stasis-dispelling efficacy.

[0054] The present Chinese medicine composition, a summary of the inventors' years of clinical experience, advocates treating hair loss by nourishing the liver and kidneys, tonifying Qi and activating blood circulation. Studies have shown that the granules can increase the expression of key proteins in the Wnt signaling pathway, reduce androgen levels, inhibit the binding of androgens to AR, thereby reducing the degradation of β-catenin and regulating the Wnt / β-catenin signaling pathway. Furthermore, the granules can inhibit cell apoptosis, prolong the hair growth cycle, and promote hair growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 This is a graph showing changes in body weight of mice in different groups in Application Example 1;

[0056] Figure 2 The area of ​​newly grown hair in the hair-removing area of ​​mice in different groups in Application Example 1;

[0057] Figure 3 This is a picture of the pathological changes of skin tissue in the hair removal area of ​​mice in different groups in Application Example 1 (HE×20);

[0058] Figure 4This is a graph showing the quantitative protein expression (×40) of Wnt10b, β-catenin, bcl-2, and BaX in the skin of mice in different groups in Application Example 1. DETAILED DESCRIPTION

[0059] To make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the following embodiments and accompanying drawings. The specific embodiments described herein are intended only to illustrate the present invention and are not intended to limit the present invention in any way. In addition, descriptions of known structures and techniques are omitted in the following description to avoid unnecessary confusion regarding the concepts of the present disclosure. Such structures and techniques are also described in many publications.

[0060] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0061] Example 1

[0062] The traditional Chinese medicine formula is: processed Polygonum multiflorum 10g, Ligustrum lucidum fruit 10g, Eclipta prostrata 10g, Polygonatum sibiricum 10g, mulberry 10g, Angelica sinensis 10g, Salvia miltiorrhiza 15g, Ligusticum chuanxiong 10g and Carthamus tinctorius 10g.

[0063] 1. Prepare the above formula into granules, wherein 5g of inulin, 0.06g of sucralose, and isomalt are added as excipients to a total amount (the yield of the paste in a small test is about 20%). The preparation is carried out according to the following steps:

[0064] (1) Pretreatment: Grind isomalt and pass through an 80-mesh sieve for later use. If the remaining materials are lumpy, pass through an 80-mesh sieve;

[0065] (2) Decoction: According to the ingredient list, weigh processed Polygonum multiflorum, Ligustrum lucidum fruit, Eclipta prostrata, Polygonatum sibiricum, mulberry, Chinese angelica, Salvia miltiorrhiza, Ligusticum chuanxiong, and Carthamus tinctorius, add 10 times the amount of water and decoct twice, the first decoction for 2 hours, the second decoction for 1 hour, filter, and set aside;

[0066] (3) Concentration: The filtrate is concentrated under reduced pressure (-0.06 to -0.10 MPa, ≤70°C) to a thick paste with a relative density of 1.15 to 1.25 (60 ± 2°C);

[0067] (4) Drying: Dry the thick paste under reduced pressure (-0.06 to -0.10 MPa, ≤70°C) until dry, grind through an 80-mesh sieve to obtain dry extract powder, and set aside;

[0068] (5) Granulation: Weigh isomalt, inulin, and dry extract powder according to the formula amount, place them together in a high-efficiency wet granulator and mix for 5 minutes to obtain a mixed powder; dissolve sucralose in 95% ethanol (about 18% of the total formula amount; the ethanol concentration and amount can be adjusted according to actual conditions) as a wetting agent to prepare a soft material, granulate into 14 mesh, dry at 55±5°C (the moisture content of the granules is controlled to be below 5%), and granulate into 14 mesh to obtain dry granules for later use;

[0069] (6) Inner packaging: composite film packaging, 10g / bag;

[0070] (7) Outer packaging: labeling, boxing, and packing, and storage after passing inspection;

[0071] (8) Inspection and warehousing: Finished products are put into storage after passing the inspection.

[0072] 2. Prepare the above formula into a decoction according to the following steps:

[0073] (1) Weighing the raw materials: Weigh the raw materials of the composition according to the formula: Polygonum multiflorum, Ligustrum lucidum fruit, Eclipta prostrata, Polygonatum sibiricum, mulberry, Angelica sinensis, Salvia miltiorrhiza, Ligusticum chuanxiong, and Carthamus tinctorius;

[0074] (2) Soaking: Put the formulated amount of raw materials into a container dedicated to Chinese medicine decoction, add 6 to 8 times the amount of cold water (drinking water that meets national health standards), and soak for 30 minutes to prepare for decoction;

[0075] (3) First decoction: Boil the soaked Chinese medicine and water over high heat until boiling, then simmer for 10-20 minutes over low heat. Turn off the heat, pour out the hot liquid and filter it to obtain 200 mL of liquid. Store in a sterilized container for later use.

[0076] (4) Second decoction: Add 6 to 8 times the amount of cold water (drinking water that meets national health standards) to the residue from the first decoction, boil over high heat until boiling, then switch to low heat and continue boiling for 10 to 20 minutes. Turn off the heat, pour out the hot liquid and filter it, and squeeze the residue to filter it, obtaining a total of 200 mL of liquid.

[0077] (5) Mixing: Pour the second decoction into the container containing the first decoction and mix evenly to obtain the decoction. Then, package the decoction in 200 mL bags.

[0078] Application Example 1

[0079] This study used a dihydrotestosterone dilution to establish an AGA mouse model and evaluated the therapeutic efficacy of the Jiuwei Shengfa Granule described in Example 1. Immunohistochemistry and real-time quantitative polymerase chain reaction (qRT-PCR) were used to quantitatively analyze the expression of key proteins and genes, including Wnt10b, β-catenin, bcl-2, and BaX, to explore the mechanism of action of Jiuwei Shengfa Granule in promoting hair regeneration and provide a reference for the clinical treatment of AGA.

[0080] 1. Materials

[0081] 1.1 Animal experiments

[0082] Sixty 6-week-old, SPF-grade C57BL / 6N male mice weighing 20-22 g were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. (License No. SYXK2023-0011). They were housed in the animal facility at the Liangxiang Campus of Beijing University of Chinese Medicine (Ethics Certificate No. BUCM-2024051303-2110) under a specific pathogen-free, SPF-grade environment with a humidity of 50% ± 10%, a temperature of 22 ± 2°C, and a 12-h light-dark cycle. They were given free access to food and water and were acclimated for 7 days. Experiments were initiated after the mice presented with no significant symptoms.

[0083] 1.2 Drugs and Reagents

[0084] Jiuwei Shengfa Granules, dihydrotestosterone injection (Beijing Solaibao Technology Co., Ltd., batch number ID0310), 4% tissue cell fixative, dimethyl sulfoxide (Beijing Solaibao Technology Co., Ltd., batch number 2306001, D8370), finasteride (Zhejiang Xianju Pharmaceutical Co., Ltd., batch number H200701112), Veet Hair Removal Cream (Reckitt Benckiser China Co., Ltd., batch number G20150999), isoflurane (Shandong Ante Animal Husbandry Technology Co., Ltd., batch number 2023090502), HRP goat anti-rabbit secondary antibody, wnt10B monoclonal recombinant antibody (Thermo Fisher Scientific, batch number GB 23303, HL2370), bcl-2, Bax, and β-catenin monoclonal recombinant antibodies (Wuhan Sanying Biotechnology Co., Ltd., batch numbers 12789-1-AP, 50599-2-Ig, and 80488-1-RR), mouse dihydrotestosterone (DHT), mouse 5α-reductase, and mouse testosterone (T) ELISA research kits (Shanghai Meike Biotechnology Co., Ltd., catalog numbers: MK3267A, RX2D234176, and MK3305A), and SteadyPure universal RNA extraction kit (Aikerui Biotechnology Co., Ltd., catalog number: AG21017).

[0085] 1.3 Instruments

[0086] Small animal anesthesia machine (Shenzhen Ruiwode Life Science Technology Co., Ltd., model R5201P), Aux electric shaver (No. 100013013621), slides and cover slips (Jiangsu Shitai Experimental Equipment Co., Ltd., models 188105 and 10212432C, respectively), microwave oven (model P70D20N1P-G5W0), pipette (model DragonBC004), atomizer pen biosharkBC004, microscope Nikon / OLYMPUS, models E100 / CX23, T100 TM Thermal Cycler (BIORAD model: 621BR45665), CFX Duet Real-Time PCR System (BIORAD model: 782B20794).

[0087] 2 Methods

[0088] 2.1 Animal grouping

[0089] The experimental animals were divided into six groups, with 10 C57 mice in each group, and the groups were randomly divided into groups using a random number table. During the experiment, mice whose skin was damaged after the first hair removal and seriously affected the experimental effect, as well as mice that died accidentally during the experiment, were removed. The groups were: normal control group, model group, finasteride group, low-, medium-, and high-dose groups of Jiuwei Shengfa Granules (the conversion coefficient between the human dose and the mouse body weight in the pharmacopoeia was 0.00026), and the low, medium, and high doses were (0.06g·kg -1 )、(0.12g·kg -1 ) and (0.24 g·kg -1 ) was conducted to evaluate the efficacy of Jiuwei Shengfa Granules at different doses. The positive control drug finasteride was administered at a dose of 0.13 mg kg -1 .

[0090] 2.2 Modeling and drug administration

[0091] After 7 days of adaptive feeding, the mice were anesthetized using a small animal anesthesia machine (4L / min). An electric shaver and a homemade plastic frame were used to shave a 2×3 cm hair removal area in the center of the mouse's back. Depilatory cream was applied to the remaining hair area, and after standing for 30 seconds, the remaining hair and depilatory cream were removed with an absorbent cotton ball. When the skin in the depilatory area was pink and there was no obvious damage, the hair follicles were judged to be in the resting phase. Mice with severe damage and scabs were eliminated. Subsequently, a dihydrotestosterone (DHT) dilution (5 mg kg -1 ) for subcutaneous injection. Since DHT is insoluble in saline, it was first dissolved in dimethyl sulfoxide to prepare 50 mg mL -1 solution, and then diluted with normal saline to 0.5 mg mL -1Subcutaneous multiple injections were performed (1 time / d). At the same time, oral administration was performed. The normal group and the model group were given equal volumes of normal saline, and the finasteride group was given finasteride (0.13 mg·kg -1 ) were administered orally. The low-, medium-, and high-dose groups of Jiuwei Shengfa Granules were given the corresponding doses of diluted solution. During the experiment, the mice were weighed and their body weights were recorded weekly. Mice were anesthetized with isoflurane (4 L / min), and photographs were taken on days 0, 7, 11, 14, 18, and 21.

[0092] 2.3 Detection indicators

[0093] 2.3.1 New hair growth area

[0094] Photographs were taken on days 0, 7, 11, 14, 18, and 21 to record the hair growth in the depilatory area of ​​the mice. Image J software was used to count and calculate the area of ​​new hair in the depilatory area on the back of each group of mice on the 21st day of the experiment.

[0095] 2.3.2 New hair length and weight

[0096] On the 21st day after modeling and drug administration, mice were anesthetized with isoflurane at a concentration of 4 L / min, their eyeballs were removed and blood samples were collected from the eyes in order to obtain skin from the back of the mice after sudden death. The six longest hairs in the new hair area on the back of the mice were selected, their lengths were measured and the average value was calculated. All new hairs were shaved off, and the remaining hair was removed with depilatory cream. The weight of the mice before and after depilation was weighed, and the difference was calculated as the weight of the new hair. The skin tissue in the depilatory area was cut into 1×1 cm 2 The samples were fixed in tissue fixative for subsequent experiments.

[0097] 2.3.3 Hematoxylin-eosin (HE) staining

[0098] Fresh tissue fixative (skin tissue from the depilated area) was fixed for at least 24 hours, flattened, and placed in a dehydrating box. The tissue was then treated sequentially with graded alcohol and xylene, dehydrated, and cleared in a dehydrator. Subsequently, the tissue was immersed in wax and embedded in an embedding machine. After the wax solidified, it was removed and trimmed into wax blocks suitable for sectioning. 4-μm-thick sections were cut using a microtome, flattened in 40°C warm water, and dried in a 60°C oven. Paraffin sections were dewaxed and rehydrated using xylene and graded alcohol. Sections were stained with hematoxylin solution, adjusted with bluing solution, and then rinsed with running water. For further staining, sections were dehydrated with graded alcohol and then immersed in eosin solution. Finally, sections were cleared with graded anhydrous ethanol and xylene and mounted with neutral gum. Hematoxylin sections were observed under a microscope, and the number and morphology of hair follicles were analyzed.

[0099] 2.3.4 Immunohistochemical assay to detect protein content in mouse skin tissue

[0100] First, sections were deparaffinized in xylene and then graded anhydrous ethanol. Sections were then immersed in PBS buffer (pH 7.4) and washed three times (3 min each time) on a decolorizing shaker. Blocking was performed with a 3% BSA solution for 20 minutes at room temperature. After removing the blocking solution, pre-prepared primary antibody solutions (Wnt10b 1:3000, β-catenin 1:500, Bcl-2 1:400, Bax 1:4000) were added dropwise and incubated overnight at 4°C. The following day, sections were washed three times with PBS, the primary antibody removed, and the secondary antibody solution added, incubating at room temperature for 50 minutes. For color development, DAB colorimetric developer was used to visualize the target area until it developed brownish-yellow. After completion, sections were rinsed with purified water. Sections were counterstained with hematoxylin solution for 3 minutes, washed with water, and then adjusted for staining depth with differentiation solution. After differentiation, sections were rinsed again and treated with a blue re-treatment to enhance contrast. Finally, dehydration and transparency were achieved by treatment with gradient anhydrous ethanol and xylene. After drying, the slides were sealed with neutral gum for microscopic observation and long-term storage.

[0101] 2.3.5 ELISA method to detect the expression levels of T, DHT and 5-AR in the skin tissue of the depilated area of ​​mice

[0102] According to the kit instructions, set up standard wells and sample wells separately. Add 50 μL of standard solution at varying concentrations to the standard wells to establish a standard curve. Add 10 μL of the sample to be tested, followed by 40 μL of diluent, to the sample wells. Add 100 μL of horseradish peroxidase (HRP) to each well, except for the blank well. After sealing, incubate the plate in a 37°C water bath for 60 minutes to allow antibody-antigen binding. After incubation, discard the reaction solution, pat the plate dry with absorbent paper, add wash solution to each well, let stand for 1 minute, then discard the solution. Repeat this wash five times to remove unbound material. After washing, add 50 μL of substrate A and 50 μL of substrate B. Substrates A and B are key components of the colorimetric reaction. Substrate A, typically hydrogen peroxide (H2O2), serves as the hydrogen acceptor, while substrate B, typically 3,3',5,5'-tetramethylbenzidine (TMB), serves as the hydrogen donor (DH2). Incubate at 37°C in the dark for 15 minutes to trigger the enzyme reaction. Finally, add 50 μL of stop solution to stop the reaction. Within 15 minutes after the reaction is terminated, measure the absorbance (OD) of each well at a wavelength of 450 nm using a microplate reader to calculate the concentration of the analyte in the sample.

[0103] 2.3.6 Real-time PCR detection of mRNA expression in mouse skin tissue

[0104] qRT-PCR RNA Extraction and Reverse Transcription: Total RNA was extracted from skin tissue using the AG Steadypure Universal RNA Kit. Following RNA extraction and concentration determination according to the kit's instructions, the reverse transcription reaction was prepared: 4 μL of 5X Evo M-MLV RT Master Mix, 2 μL of RNA sample (1000 ng), and the volume was made up to 20 μL with enzyme-free water. The reverse transcription reaction was performed at 37°C for 15 minutes, followed by heating to 85°C for 5 seconds, and finally incubated at 4°C for 3 minutes to complete cDNA synthesis. For qPCR amplification, the reaction system was supplemented with 10 μL of 2X SYBR Green, 0.4 μL of each forward and reverse primer, 2 μL of cDNA sample, and 7.2 μL of enzyme-free water. The PCR amplification program was as follows: initial denaturation at 95°C for 3 minutes, followed by 39 cycles of denaturation at 95°C for 10 seconds, annealing at 60°C for 30 seconds, and extension for 30 seconds. Results: Quantification was performed based on the CT values ​​of the qPCR results, and relative expression levels were calculated. Primer information was synthesized by Hunan Aikerui Bioengineering Co., Ltd. (see Table 1).

[0105] Table 1 Primer sequences

[0106]

[0107] 2.9 Statistical Methods

[0108] ImageJ and SPSS 27.0 software were used to perform statistical analysis of experimental data. All data were expressed as mean ± standard deviation (mean ± SD) to ensure data accuracy. In the comparison of multiple groups of data, the first step was to test for homogeneity of variance. If the test was met, a one-way analysis of variance (ANOVA) was performed. When the ANOVA results were significant, pairwise comparisons were performed using the least significant difference (LSD) method. For data with unequal variance, the Dunnett T test was used for analysis. Statistical significance was set at P < 0.05.

[0109] 3 Results

[0110] 3.1 Mouse weight

[0111] Changes in the weight of mice in different groups During the entire experimental period, as the experimental time increased, the weight of mice in each group (6 mice in each group) increased slowly, proving that the mice were in good growth condition during the experiment. Figure 1 .

[0112] 3.2 New hair growth area within the hair loss region of mice in different groups

[0113] Compared with the normal group, the area of ​​new hair in the hair loss area of ​​the model group mice was significantly reduced (P<0.01), indicating that subcutaneous dihydrotestosterone can successfully induce the AGA model; compared with the model group, the area of ​​new hair in the hair loss area of ​​the mice in the low, medium and high dose groups of Jiuwei Shengfa Granules were significantly increased (P<0.01). Although the area of ​​new hair in the finasteride group and the high dose group of Jiuwei Shengfa Granules showed an upward trend, it did not reach statistical significance. The above results show that Jiuwei Shengfa Granules can effectively alleviate the hair loss phenomenon caused by male hormones. See Table 2 and Figure 2 .

[0114] Table 2 New hair area in different mouse hair removal areas ( n=6)

[0115]

[0116] Note: Compared with the normal group, a: P < 0.05, b: P < 0.01; compared with the model group, c: P < 0.05, d: P < 0.01.

[0117] 3.3 Changes in the weight, length, and number of hair follicles of newly grown hair in the hair removal area of ​​mice in different groups

[0118] Compared with the normal group, the length, weight and unit area of ​​new hair in the hair loss area of ​​the model group mice were significantly decreased (P<0.05, P<0.01). Compared with the model group, the length of new hair in the low, medium and high dose groups of Jiuwei Shengfa Granules and the finasteride group was significantly increased (P<0.01). The hair weight of mice in the low and medium dose groups of Jiuwei Shengfa Granules was significantly increased (P<0.05), and the hair weight in the high dose group showed an increasing trend, but it was not statistically significant. HE results showed that compared with the normal group, the number of hair follicles in the model group mice was significantly reduced (P<0.01), and the arrangement was disordered, the hair morphology was irregular and poorly developed; compared with the model group, the number of hair follicles in each drug-treated group was significantly increased (P<0.01), and the arrangement was neat, the morphology was regular, and the development was good. In summary, Jiuwei Shengfa Granules can improve the reduction in hair length, weight and number of hair follicles in AGA mice induced by subcutaneous injection of dihydrotestosterone. See Table 3 and Figure 3 .

[0119] Table 3 Length of new hair and number of hair follicles in the hair removal area of ​​mice in different groups ( n=6)

[0120]

[0121]

[0122] Note: Compared with the normal group, a: P < 0.05, b: P < 0.01; compared with the model group, c: P < 0.05, d: P < 0.01.

[0123] 3.4 ELISA results showed the expression levels of T, DHT and 5-AR in the skin tissue of the depilated area of ​​mice in different groups

[0124] Compared with the normal group, the levels of testosterone (T) and dihydrotestosterone (DHT) in the hairless skin of mice in the model group were significantly increased (P < 0.05, P < 0.01). Compared with the model group, the levels of T and DHT in the hairless skin of mice in the low- and medium-dose Jiuwei Shengfa Granule groups were significantly decreased (P < 0.01, P < 0.05). Although there was a downward trend in the high-dose group, it was not statistically significant. Regarding 5-α-reductase (5-AR), no significant changes were observed in the model group compared with the normal group. However, compared with the model group, the 5-AR levels in the skin of mice in the low-, medium-, and high-dose Jiuwei Shengfa Granule groups were significantly decreased (P < 0.01). These results indicate that Jiuwei Shengfa Granule can effectively inhibit the DHT-induced increase in androgen levels in mouse skin. Specific data are shown in Table 4.

[0125] Table 4 T, DHT and 5-AR contents in skin tissue of depilatory area of ​​mice in different groups ( n=6)

[0126]

[0127] Note: Compared with the normal group, a: P < 0.05, b: P < 0.01; compared with the model group, c: P < 0.05, d: P < 0.01.

[0128] 3.5 Expression levels of Wnt10b, β-catenin, BaX, and bcl-2 proteins

[0129] The results of immunohistochemical analysis showed that compared with the normal group, the expression of Wnt10b, β-catenin and bcl-2 proteins in the skin of the hair loss area of ​​the model group mice was significantly decreased (P<0.01), while the expression of BaX protein was significantly increased (P<0.01). Compared with the model group, the expression of Wnt10b, β-catenin and bcl-2 proteins in the skin of mice in each dose group of Jiuwei Shengfa Granules was significantly increased (P<0.01), and the expression of BaX protein was significantly decreased (P<0.01). The above results indicate that Jiuwei Shengfa Granules may promote hair regeneration in AGA mice by upregulating the expression of Wnt10b, β-catenin and bcl-2 proteins and downregulating the expression of BaX protein, activating the Wnt / β-catenin signaling pathway and inhibiting cell apoptosis. See Table 5 and Figure 4 .

[0130] Table 5 Expression levels of Wnt10b, β-catenin, bcl-2 and BaX proteins in the skin tissue of the depilated area of ​​mice in different groups ( n=3)

[0131]

[0132] Note: Compared with the normal group a: P < 0.05, b: P < 0.01; compared with the model group c: P < 0.05, d: P < 0.01

[0133] 3.6 Expression levels of Wnt10b, β-catenin, BaX, and bcl-2 mRNA in different groups

[0134] qRT-PCR results: Compared with the normal group, the expression of Wnt10b, β-catenin, and bcl-2 genes in the skin of mice in the model group was significantly decreased (P < 0.01), while the expression of BaX gene was significantly increased (P < 0.01). Compared with the model group, the expression of Wnt10b, β-catenin, and bcl-2 genes in the hair loss area of ​​mice in all dose groups of Jiuwei Shengfa Granule was significantly increased (P < 0.01), while the expression of BaX gene was significantly decreased (P < 0.01). These results suggest that Jiuwei Shengfa Granule may promote hair regeneration in AGA mice by upregulating the expression of Wnt10b, β-catenin, and bcl-2 genes and downregulating the expression of BaX gene, thereby activating the Wnt / β-catenin signaling pathway and inhibiting cell apoptosis. See Table 6.

[0135] Table 6 Gene contents of Wnt10b, β-catenin, bcl-2 and BaX in skin tissues of mice in different groups ( n=3)

[0136]

[0137] Note: Compared with the normal group, a: P < 0.05, b: P < 0.01; compared with the model group, c: P < 0.05, d: P < 0.01.

[0138] 4 Discussions

[0139] The occurrence of AGA is related to androgen levels, genetic factors, hemorheology, and lifestyle. Testosterone regulates the interaction between hair follicle epithelial cells and endothelial mesenchymal cells, thereby affecting hair growth rate, dermal papilla size, dermal papilla cell activity, keratinocyte activity, and melanocyte activity. The pathogenesis of AGA is closely related to the effects of the androgen receptor (AR) and dihydrotestosterone (DHT). Type II 5α-reductase converts testosterone (T) to DHT in the bloodstream, and DHT binds to the androgen receptor with five times the affinity of testosterone. These hormone isomers play a key role in androgen metabolism. Although male patients have normal androgen levels, increased 5α-reductase activity leads to elevated DHT levels, which in turn causes hair loss. The results of this study showed that compared with the model group, Jiuwei Shengfa Granule significantly reduced DHT and 5-AR levels in the skin of mice with hair loss (P < 0.01).

[0140] The Wnt signaling pathway primarily consists of the canonical Wnt / β-catenin pathway and the non-canonical Wnt / polarity pathway. The Wnt / β-catenin pathway plays a key role in hair follicle developmental matrix cells (DPCs) in inducing hair growth and in the transition of hair follicles from telogen to anagen. Dihydrotestosterone (DHT) competitively binds to the androgen receptor (AR), forming a complex that activates glycogen synthase kinase 3β (GSK-3β) in the canonical Wnt / β-catenin pathway. Activation of GSK-3β promotes the phosphorylation of β-catenin, leading to its ubiquitination and proteasomal degradation. This process inhibits the differentiation of hair follicle stem cells in patients with androgenic alopecia (AGA). DHT stimulation inhibits the expression of the Wnt agonist Wnt10b while increasing the levels of the Wnt antagonist DKK1. As a catagen-inducing factor, DKK1 blocks the activation of the canonical Wnt / β-catenin signaling pathway by binding to Frizzled, thereby inducing the expression of the pro-apoptotic protein Bax. This process ultimately leads to apoptosis of outer root sheath keratinocytes, interfering with the differentiation and proliferation of hair follicle stem cells. Wnt10b is a unique ligand in the Wnt signaling pathway. It promotes hair follicle regeneration and hair growth through the canonical Wnt / β-catenin signaling pathway, prolonging the anagen phase. When β-catenin levels are elevated, it effectively activates stem cell proliferation and promotes hair follicle regeneration. This process accelerates the differentiation of hair follicle stem cells into hair follicles, thereby activating the hair growth cycle, shortening the telogen phase, and promoting a rapid transition to the anagen phase. Ultimately, hair maintains a state of continuous growth, leading to increased hair length and weight. Research on skin hair follicles has shown that Bcl-2 and Bax, members of the Bcl-2 protein family, play important roles in hair follicle cell apoptosis. Bcl-2, an inhibitor of apoptosis, and Bax, a pro-apoptotic factor, have been shown to correlate closely with the timing of hair follicle regeneration in cashmere goats. The Bcl-2 / Bax ratio is low during the anagen phase and significantly increases during the catagen and telogen phases. Previous studies have shown that plants can effectively inhibit apoptosis in HDPC cells by increasing the ratio of anti-apoptotic to pro-apoptotic Bcl-2 and Bax. Furthermore, Bax and Bcl-2 can independently regulate apoptosis in vivo. While topical minoxidil solution and oral finasteride are widely used for the treatment of hair loss, their significant adverse reactions necessitate the development of safer and more effective treatments. The results of this study showed that the expression levels of Wnt10b and β-catenin proteins and their genes in the hair loss areas of mice treated with Jiuwei Shengfa Granules were significantly higher than those in the model group (P < 0.01).In addition, the protein and gene expression levels of apoptosis inhibitor Bcl-2 were significantly increased, while the protein and gene expression levels of apoptosis initiator Bax were significantly decreased (P<0.01).

[0141] AGA is closely related to the state of qi, blood, and essence in the human body, as well as the functions of the liver, spleen, and kidney. Its main pathogenesis includes liver and kidney deficiency, liver depression and blood stasis, internal accumulation of deficiency dampness and heat, and blood heat turning into dryness.

[27] . Jiuwei Shengfa Granules takes nourishing the liver and kidneys, replenishing qi and activating blood circulation as its treatment principle, and explores the treatment plan for androgenic alopecia from the perspective of the liver and kidneys. Traditional Chinese medicine has few side effects in treating AGA, high patient compliance, and a wide spectrum of activity. It can regulate the Wnt / β-catenin signaling pathway through multiple pathways and multiple targets, thereby effectively inhibiting the occurrence of AGA and achieving therapeutic effects. The results of this study showed that Jiuwei Shengfa Granules can significantly increase the expression levels of Wnt10b, β-catenin and Bcl-2 in mouse skin, and significantly reduce the expression of Bax. These results suggest that Jiuwei Shengfa Granules exerts its hair growth effect by promoting the conduction of the Wnt / β-catenin signaling pathway, and slows down hair loss by inhibiting cell apoptosis.

[0142] In summary, this study used a dihydrotestosterone (DHT) dilution to establish an androgenic alopecia (AGA) mouse model. The efficacy of Jiuwei Shengfa Granules in promoting hair regeneration in AGA mice was validated by assessing the area, length, and weight of newly formed hair, as well as the number of hair follicles in local skin tissue. The results showed that the number of hair follicles in the treated group was significantly higher than that in the model group, with follicles of uniform size and good development. In contrast, the number of hair follicles in the model group was reduced, with irregular arrangement and gradual atrophy and shrinkage. Furthermore, further analysis of testosterone (T), dihydrotestosterone (DHT), and 5α-reductase levels in the mouse skin revealed that Jiuwei Shengfa Granules significantly reduced androgen levels and 5α-reductase levels in the local skin, thereby reducing hair loss. Immunohistochemistry and QRT-PCR analyses confirmed the expression levels of key proteins in the Wnt / β-catenin signaling pathway, as well as bcl-2 and BaX. These results suggest that Jiuwei Shengfa Granules may have a significant therapeutic effect on AGA mice by activating the Wnt / β-catenin signaling pathway and inhibiting cell apoptosis. This granule not only embodies the holistic concept and principle of syndrome differentiation and treatment in Traditional Chinese Medicine, but has also been validated by modern pharmacological research for its scientificity and effectiveness, providing a new treatment option for AGA patients. However, additional clinical trials with larger sample sizes are needed to further confirm the efficacy and mechanism of Jiuwei Shengfa Granule on hair growth in AGA patients.

[0143] Application Example 2 Clinical efficacy of the present invention

[0144] The present invention treated 35 patients with hair loss in the outpatient clinic of the National Medical Hall, aged 25-55 years, including 27 males and 8 females, and took the Nine-Ingredient Hair Growth Granules prepared in Example 1, three times a day, 10 grams each time; and achieved relatively satisfactory results, with 15 cases showing significant effects, 16 cases showing effective effects, and 4 cases showing ineffective effects (2 of which did not continue the treatment), with a total effective rate of 88.57%.

[0145] After 30 days of treatment, hair loss is generally reduced or eliminated, dandruff and oil secretion are significantly reduced, and white hair begins to appear in the alopecia area. After 60 days of treatment, the white hair of alopecia patients turns gray-black, and hair grows about 1-2 centimeters. For patients with seborrheic alopecia, the fine hair grows coarser and stronger, and new hair begins to grow through the skin. After 90 days of treatment, the patient's hair growth is essentially back to normal.

[0146] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for treating hair loss, wherein the raw materials are in the following weight ratios: 1-50 parts of processed Polygonum multiflorum, 1-50 parts of Ligustrum lucidum fruit, 1-50 parts of Eclipta prostrata, 1-50 parts of Polygonatum sibiricum, 1-50 parts of mulberry, 1-50 parts of Angelica sinensis, 5-100 parts of Salvia miltiorrhiza, 1-50 parts of Chuanxiong and 1-50 parts of Carthamus tinctorius.

2. The Chinese medicine composition according to claim 1, characterized in that The raw materials are prepared in the following proportions by weight: 5-20 parts of Radix Polygoni Multiflori, 5-20 parts of Fructus Ligustri Lucidi, 5-20 parts of Herba Ecliptae, 5-20 parts of Rhizoma Polygonati, 5-20 parts of Mulberry, 5-20 parts of Radix Angelicae Sinensis, 10-30 parts of Radix Salviae Miltiorrhizae, 5-20 parts of Rhizoma Chuanxiong and 5-20 parts of Carthamus Tinctorius; Preferably, the weight ratio of the Chinese medicinal raw materials is: 8-12 parts of processed Polygonum multiflorum, 8-12 parts of Ligustrum lucidum fruit, 8-12 parts of Eclipta prostrata, 8-12 parts of Polygonatum sibiricum, 8-12 parts of mulberry, 8-12 parts of Angelica sinensis, 12-17 parts of Salvia miltiorrhiza, 8-12 parts of Chuanxiong and 8-12 parts of Carthamus tinctorius.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that The weight ratio of the Chinese medicinal materials is as follows: 10 parts of processed Polygonum multiflorum, 10 parts of Ligustrum lucidum fruit, 10 parts of Eclipta prostrata, 10 parts of Polygonatum sibiricum, 10 parts of mulberry, 10 parts of Angelica sinensis, 15 parts of Salvia miltiorrhiza, 10 parts of Chuanxiong and 10 parts of Carthamus tinctorius.

4. A Chinese medicine preparation comprising the Chinese medicine composition according to any one of claims 1 to 3.

5. The Chinese medicine preparation according to claim 4, characterized in that The dosage form of the traditional Chinese medicine preparation is selected from one or more of granules, decoctions, tablets, lotions, ointments, creams, hydrogels or essential oils.

6. The Chinese medicine preparation according to claim 4, characterized in that The dosage form of the preparation is granules and / or decoction.

7. Use of the traditional Chinese medicine composition according to any one of claims 1 to 3 or the traditional Chinese medicine preparation according to any one of claims 4 to 6 in the preparation of a medicament for treating or preventing hair loss.

8. The use according to claim 7, characterized in that The hair loss is androgenic alopecia.

9. Use of the traditional Chinese medicine composition according to any one of claims 1 to 3 or the traditional Chinese medicine preparation according to any one of claims 4 to 6 in a product for promoting hair growth.

10. Use of the traditional Chinese medicine composition according to any one of claims 1 to 3 or the traditional Chinese medicine preparation according to any one of claims 4 to 6 in the preparation of a drug for lowering androgen levels, inhibiting 5-AR, promoting Wnt / β-catenin signaling pathway activity and / or inhibiting cell apoptosis.

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