Hair-nourishing traditional Chinese medicine composition for preventing and treating alopecia

Abstract

The invention discloses a hair-nourishing traditional Chinese medicine composition for preventing and treating alopecia. The hair-nourishing traditional Chinese medicine composition is prepared from the following raw materials: astragalus membranaceus, ginseng, glossy privet fruit, rhizoma drynariae, fructus evodiae, salvia miltiorrhiza, polygonum multiflorum, angelica sinensis, radix rehmanniae recen, prepared rehmannia root, semen coicis, poria cocos and bighead atractylodes rhizome. The invention provides a multi-component, multi-target and multi-channel comprehensive solution aiming at the core pathogenesis of alopecia through the combination of the traditional Chinese medicine holistic view and the modern pharmacology, overcomes the defects of the existing product, and opens up a safer and systematic health care path for alopecia prevention and treatment.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, and particularly relates to a hair-nourishing traditional Chinese medicine composition for preventing and treating hair loss. Background Art

[0002] Hair loss refers to the phenomenon of hair shedding. The normally shed hairs are all hairs in the telogen and exogen phases. Since the hairs entering the telogen phase and the newly entering anagen phase are constantly in a dynamic balance, the normal number of hairs can be maintained. Pathological hair loss is abnormal or excessive hair shedding, often caused by factors such as genetics, mental stress, and endocrine disorders, leading to damaged hair follicles and disrupted hair growth cycles.

[0003] In the current field of hair loss treatment, although there are various treatment methods and products, each method faces its unique challenges and limitations. First, minoxidil, as a widely used topical treatment drug, has certain efficacy for hair loss diseases such as male and female androgenetic alopecia and alopecia areata. However, long-term use of minoxidil may cause a series of side effects, including but not limited to scalp irritation, hirsutism (i.e., hair growth in unexpected locations), etc. In addition, although minoxidil can promote hair regrowth, its mechanism of action has not been fully clarified, which limits its effectiveness as a single therapy. On the other hand, finasteride is an oral drug designed specifically for men. By inhibiting 5α-reductase, it reduces the production of dihydrotestosterone (DHT), thereby slowing down the hair loss process and promoting hair regrowth. Although finasteride is effective for many patients, it may also bring a series of side effects, such as sexual dysfunction, hormonal level changes, etc. It is worth noting that finasteride is not suitable for female patients, which further limits its scope of application. Anti-hair loss shampoos usually contain active ingredients such as ketoconazole or caffeine. These ingredients can effectively control sebum secretion on the scalp, provide short-term oil control effects, and may help improve the scalp environment. However, long-term use of such products may lead to an imbalance in the scalp microecology and affect scalp health. In addition, the efficacy of these topical products mainly focuses on the surface level and cannot deeply address the root causes of hair loss. Traditional Chinese medicine compound prescriptions, as another treatment option, rely on traditional Chinese medicine theory to regulate the internal environment of the body to achieve the purpose of improving hair loss. Although some traditional Chinese medicine formulas have shown certain efficacy, due to the lack of modern pharmacological verification, their exact mechanism of action and safety are still unclear. In addition, since traditional Chinese medicine compound prescriptions are often adjusted based on individual differences, it is difficult to formulate standardized treatment plans, which also limits their large-scale application. Summary of the Invention

[0004] The purpose of the present invention is to provide a hair-nourishing traditional Chinese medicine composition for preventing and treating hair loss, in order to put forward health care ideas and theoretical bases for hair loss.

[0005] In order to achieve the above purpose, the technical solution adopted by the present invention is as follows: A traditional Chinese medicine composition for preventing and treating hair loss and nourishing hair is prepared from raw materials in the following parts by weight: 10-50 parts of astragalus membranaceus, 100-200 parts of ginseng, 100-200 parts of glossy privet fruit, 100-200 parts of drynaria rhizome, 3-50 parts of evodia rutaecarpa, 100-200 parts of salvia miltiorrhiza, 5-50 parts of polygonum multiflorum, 5-50 parts of angelica sinensis, 5-50 parts of raw rehmannia root, 5-50 parts of prepared rehmannia root, 3-50 parts of coix seed, 5-50 parts of poria cocos, and 3-50 parts of atractylodes macrocephala.

[0006] Preferably, the parts by weight of each raw material are: 10 parts of astragalus membranaceus, 158 parts of ginseng, 158 parts of glossy privet fruit, 158 parts of drynaria rhizome, 9 parts of evodia rutaecarpa, 158 parts of salvia miltiorrhiza, 15 parts of polygonum multiflorum, 15 parts of angelica sinensis, 15 parts of raw rehmannia root, 12 parts of prepared rehmannia root, 6 parts of coix seed, 15 parts of poria cocos, and 30 parts of atractylodes macrocephala.

[0007] The dosage forms of the traditional Chinese medicine composition for nourishing hair of the present invention include decoction, powder or ointment prepared from the raw materials in the above parts by weight.

[0008] The beneficial effects of the present invention compared with the prior art are as follows: Traditional Chinese medicine has a long history of understanding the growth and loss of hair. "Plain Questions · Chapter on the Genuine Conformation of Ancient People" states that "at the age of seven for girls, the kidney qi is strong, the teeth change and the hair grows... at the age of twenty-eight, the bones and tendons are strong, the hair grows to its fullest, and the body is strong and healthy... at the age of eight for men, the kidney qi is full, the hair grows and the teeth change... at the age of forty, the kidney qi declines, the hair falls out and the teeth become withered", which details the prosperity and decline evolution of hair with age. Traditional Chinese medicine treatment of seborrheic alopecia has the advantages of multiple targets, comprehensive consideration and individualized treatment. By regulating the internal organs of the five zang-organs, not only the purpose of treating seborrheic alopecia can be achieved, but also great benefits can be obtained for other parts of the body, and a healthier state can be maintained at a deeper level. The traditional Chinese medicine compound approved by the patent has a high recognition and universality. A study sorted out the drug compatibility in 44 patents on the treatment of alopecia with traditional Chinese medicine and found that the properties of the drugs are mainly cold and flat, the taste is most commonly sweet, the meridian tropism is mainly the liver, kidney and spleen meridians, and the efficacy is mainly nourishing the liver and kidney and clearing heat and detoxifying. However, the efficacy of traditional Chinese medicine compounds is usually the result of the combined action of multiple active ingredients in the drugs. Therefore, the complete identification of all active ingredients in traditional Chinese medicine compounds is the premise and basis for establishing a quality control system and clarifying the material basis of drug efficacy.

[0009] Ginseng, Glossy Privet Fruit and Drynaria Rhizome are the sovereign drugs in the traditional Chinese medicine composition of the present invention. Ginseng tastes sweet and slightly bitter, and is slightly warm in nature, entering the spleen, lung, heart and kidney meridians, having the effects of greatly replenishing vital energy, tonifying the heart qi, calming the mind and improving intelligence. Hair loss is often due to insufficient qi and blood and deficiency of essence and blood. By greatly replenishing vital energy, ginseng can effectively enhance the qi and blood biochemistry function of the body, promote the generation of essence and blood, and then nourish the hair roots, enabling the hair to obtain sufficient nutrition and playing a role in strengthening hair and promoting hair growth; Glossy Privet Fruit is slightly cold in nature, tastes sweet and bitter, and enters the liver and kidney meridians, having the effects of nourishing the liver and kidney, improving eyesight and blackening hair. Hair loss is mostly due to yin deficiency of the liver and kidney. By nourishing the liver and kidney, Glossy Privet Fruit can effectively improve the condition of yin deficiency of the liver and kidney, enhance the nourishing function of essence and blood of the liver and kidney, enable the hair to be fully nourished, and thus play a role in strengthening hair and promoting hair growth; Drynaria Rhizome is warm in nature, tastes bitter and sweet, and enters the liver and kidney meridians, having the effects of tonifying the kidney and strengthening bones, and relieving pain from fractures. Hair loss is related to insufficient kidney qi. By tonifying the kidney and strengthening bones, Drynaria Rhizome can effectively enhance kidney qi, stabilize hair roots and prevent hair loss.

[0010] Astragalus membranaceus, Salvia miltiorrhiza and Polygonum multiflorum Thunb. are the ministerial drugs in the traditional Chinese medicine composition of the present invention. Astragalus membranaceus tastes sweet and is slightly warm in nature, entering the spleen and lung meridians, having the effects of tonifying qi and lifting yang, securing the exterior and stopping sweating, promoting the production of body fluid and nourishing blood. Hair loss is often due to qi deficiency failing to secure and hold hair roots. By tonifying qi and lifting yang, Astragalus membranaceus can effectively enhance the securing and holding function of the body, stabilize hair roots and prevent hair loss; its effect of promoting the production of body fluid and nourishing blood can also provide sufficient nutrition for hair growth; Salvia miltiorrhiza tastes bitter and is slightly cold in nature, entering the heart and liver meridians, having the effects of promoting blood circulation to remove blood stasis, clearing the heart and relieving vexation. Hair loss is related to blood stasis obstruction and poor qi and blood circulation. By promoting blood circulation to remove blood stasis, Salvia miltiorrhiza can effectively improve the blood circulation of the scalp, promote qi and blood circulation, and provide good blood supply for hair growth; Polygonum multiflorum Thunb. tastes bitter, sweet and astringent, and is slightly warm in nature, entering the liver and kidney meridians, having the effects of tonifying the liver and kidney, benefiting essence and blood, and blackening hair and beard. Hair loss is mostly due to deficiency of the liver and kidney and deficiency of essence and blood. By tonifying the liver and kidney and benefiting essence and blood, Polygonum multiflorum Thunb. can effectively nourish hair roots, enable the hair to obtain sufficient nutrition, and play a role in strengthening hair and promoting hair growth.

[0011] Angelica sinensis, Rehmannia glutinosa, Rehmannia preparata, and Coix lacryma-jobi are the adjuvant drugs in the traditional Chinese medicine composition of the present invention. Angelica sinensis is sweet, pungent, and warm in nature, and belongs to the liver, heart, and spleen meridians. It has the effects of enriching blood and promoting blood circulation, regulating menstruation and relieving pain. Hair loss is related to the inability of blood deficiency to nourish the hair. Angelica sinensis can effectively improve the condition of blood deficiency by enriching blood and promoting blood circulation, providing sufficient blood nourishment for hair growth; Rehmannia glutinosa is sweet, bitter, and cold in nature, and belongs to the heart, liver, and kidney meridians. It has the effects of clearing heat and cooling blood, nourishing yin and promoting fluid production; Rehmannia preparata is sweet and slightly warm in nature, and belongs to the liver and kidney meridians. It has the effect of nourishing yin and enriching blood. Hair loss is often caused by yin deficiency and blood heat, and deficiency of essence and blood. Rehmannia glutinosa and Rehmannia preparata are used in combination, which can not only clear heat and cool blood, nourish yin and promote fluid production, but also nourish yin and enrich blood, jointly nourishing the hair roots and enabling the hair to be fully nourished; Coix lacryma-jobi is sweet, light, and cool in nature, and belongs to the spleen, stomach, and lung meridians. It has the effects of promoting diuresis to eliminate dampness, strengthening the spleen and stopping diarrhea. Hair loss is related to the accumulation of damp pathogen and spleen deficiency with excessive dampness. Coix lacryma-jobi can effectively resolve the damp pathogen in the body and improve the condition of spleen deficiency with excessive dampness by promoting diuresis to eliminate dampness, thus creating a good internal environment for hair growth.

[0012] Poria cocos, Atractylodes macrocephala, and Evodia rutaecarpa are the guiding drugs in the traditional Chinese medicine composition of the present invention. Poria cocos is sweet, light, and neutral in nature, and belongs to the heart, lung, spleen, and kidney meridians. It has the effects of promoting diuresis to eliminate dampness, strengthening the spleen and calming the mind. Hair loss is related to the accumulation of damp pathogen and spleen deficiency with excessive dampness. Poria cocos can effectively resolve the damp pathogen in the body and improve the condition of spleen deficiency with excessive dampness by promoting diuresis to eliminate dampness. At the same time, its effect of strengthening the spleen and calming the mind can also enhance the transportation and transformation function of the spleen and stomach, providing sufficient nutrition for hair growth; Atractylodes macrocephala is sweet, bitter, and warm in nature, and belongs to the spleen and stomach meridians. It has the effects of strengthening the spleen and replenishing qi, drying dampness and promoting diuresis. Hair loss is related to spleen deficiency with excessive dampness and qi and blood deficiency. Atractylodes macrocephala can effectively enhance the transportation and transformation function of the spleen and stomach and promote the generation of qi and blood by strengthening the spleen and replenishing qi. At the same time, its effect of drying dampness and promoting diuresis can also resolve the damp pathogen in the body and improve the condition of spleen deficiency with excessive dampness; Evodia rutaecarpa is pungent, bitter, and hot in nature, and belongs to the liver, spleen, stomach, and kidney meridians. It has the effects of warming the middle-jiao to stop pain and dispersing the stagnated liver qi downward. Hair loss is related to liver qi stagnation and qi and blood stagnation. Evodia rutaecarpa can effectively relieve symptoms such as chest tightness and emotional discomfort caused by liver qi stagnation by dispersing the stagnated liver qi downward. At the same time, its effect of warming the middle-jiao to stop pain can also improve the condition of spleen and stomach yang deficiency and promote the transportation and transformation function of the spleen and stomach.

[0013] The traditional Chinese medicine composition of the present invention takes ginseng, glossy privet fruit, and drynaria rhizome as the main drugs, supplemented by astragalus membranaceus, salvia miltiorrhiza, polygonum multiflorum, etc., which tonify qi and nourish blood, promote blood circulation to remove blood stasis, and nourish the liver and kidney, jointly nourishing hair follicles and stabilizing the root of hair; assisted by angelica sinensis, rehmannia glutinosa, prepared rehmannia root, coix seed, etc., which enrich blood and nourish yin, promote diuresis and percolate dampness, further improving the condition of qi and blood deficiency and internal accumulation of damp pathogen, providing sufficient nutrition and a good internal environment for hair growth; and made with poria cocos, atractylodes macrocephala, evodia rutaecarpa, etc., which strengthen the spleen and replenish qi, dry dampness and promote diuresis, and soothe the liver and lower qi, enhancing the transportation and transformation function of the spleen and stomach, resolving damp pathogen in the body, improving the condition of liver qi stagnation, promoting qi and blood circulation, and enabling hair to be fully nourished. The whole formula exerts the effects of tonifying qi and nourishing blood, nourishing the liver and kidney, promoting blood circulation to remove blood stasis, promoting diuresis and percolating dampness, and soothing the liver and lower qi, is applicable to various types of hair loss problems, can effectively nourish hair follicles, play a role in strengthening hair and promoting hair growth, and improve hair loss symptoms.

[0014] Based on traditional Chinese medicine theory, the present invention explores natural products for hair follicle maintenance from traditional Chinese medicine herbs, establishes a professional commercial hair problem detection system, determines the key proteins it triggers, and further verifies the effectiveness and molecular mechanism of the traditional Chinese medicine composition through animal experiments, using more scientific and effective research results products to solve hair problems for consumers.

[0015] The present invention screens and summarizes the active ingredients and potential action targets of each medicinal flavor in the traditional Chinese medicine composition proposed by the present invention through the TCMSP / BATMAN / HERB / ETCM traditional Chinese medicine databases and analysis platforms, retrieves the summary of disease-related targets in the databases OpenTargets / OMIM / GeneCards, constructs a compound target-disease Venn diagram, and obtains a list of intersection targets between drug targets and disease targets; uses Cytoscape3.9.1 software to draw a drug-ingredient-target-disease network diagram. Constructs a protein-protein interaction PPI network diagram through the STRING database and Cytoscape to screen the core targets among the intersection targets. Uses the clusterProfiler package of the R 4.2.1 software to perform GO (gene ontology) function and KEGG (Kyoto encyclopedia of genes and genomes) pathway enrichment analysis to explore the potential action mechanism of the hair-nourishing traditional Chinese medicine composition in the present invention on hair loss, and constructs a drug-ingredient-target-pathway network diagram with Cytoscape; finally, uses AutoDock Tools to perform molecular docking on the key active ingredients and core targets to verify the results of network pharmacology analysis.

[0016] Through oral bioavailability OB and drug-like property parameter DL, 282 active ingredients, 2214 active ingredient targets, 410 hair loss-related targets and 113 intersection targets were screened out in the hair care Chinese medicine composition; GO functional enrichment analysis obtained 1518 biological process BP entries, 8 cell composition CC entries, and 33 molecular function MF entries (p.adjust<0.01); KEGG pathway enrichment analysis screened 142 (P<0.05) significant signal pathways, mainly involving Inflammatory boweldisease, Th17 cell differentiation, JAK-STAT signaling pathway, Graft-versus-hostdisease, Influenza A, Allograft rejection, Cytokine-cytokine receptorinteraction, Type I diabetesmellitus, Rheumatoid arthritis, Hepatitis B, AGE-RAGEsignaling pathway in diabetic, Proteoglycans incancer, Chagas disease and other signal pathways; molecular docking showed that the TOP active ingredients MOL010861 and MOL008691 screened out in the drug had good affinity with the screened core target CTNNB1. MOL008691 had good affinity with the screened core target IFNG. MOL000358, MOL010861 and MOL008691 all had good affinity with the screened core target IL-10.

[0017] The present invention solves the complex pathogenesis of hair loss through multiple synergistic approaches such as invigorating qi and nourishing blood (ginseng, astragalus), nourishing the liver and kidney (ligustrum lucidum, polygonum multiflorum), promoting blood circulation and removing blood stasis (salvia miltiorrhiza), and promoting dampness and strengthening the spleen (coix seed, atractylodes macrocephala); network pharmacology confirms that the composition can regulate 113 intersection targets and 142 pathways (such as JAK-STAT, inflammatory pathways), covering multiple dimensions such as hair follicle cycle, immune regulation, and metabolic balance; avoids the side effects of hormonal intervention of Western medicine, gently regulates through natural ingredients, strictly controls the dosage of polygonum multiflorum (5-50 parts) in the formula, and is combined with detoxifying drugs such as rehmannia root to reduce the risk of liver toxicity; provides a variety of forms such as decoctions, powders, and ointments to suit different body constitutions; uses molecular docking and animal experiments to verify, breaking through the limitation of traditional Chinese medicine that is "based on experience"; not only improves the symptoms of hair loss, but also regulates overall health, which is in line with the concept of "preventive treatment" in traditional Chinese medicine.

[0018] Through the research on the mechanism of hair loss, it is confirmed that the traditional Chinese medicine composition proposed in the present invention contains a variety of active ingredients, which can act on multiple key targets related to hair loss, and may further play a role in treating diseases synergistically by participating in the regulation of multiple pathways such as [pathway name]. This research provides a basis for further verifying the related targets and pathways of the hair-nourishing traditional Chinese medicine composition in preventing and treating hair loss, predicts the possible active ingredients of the hair-nourishing traditional Chinese medicine composition, and conducts an effectiveness study on its effectiveness through animal experiments, providing a theoretical basis and new ideas for further research on the health care of hair loss. By combining the overall concept of traditional Chinese medicine with modern pharmacology, the present invention provides a comprehensive solution with multiple components, multiple targets, and multiple pathways for the core pathogenesis of hair loss, making up for the defects of existing products and opening up a safer and more systematic health care path for the prevention and treatment of hair loss. Description of the Drawings

[0019] Figure 1 It is the intersection diagram of active ingredients between different databases in Experimental Example 1 of the present invention; Figure 2 It is the intersection diagram of targets between different databases in Experimental Example 1 of the present invention; Figure 3 It is the Venn diagram of the hair-nourishing traditional Chinese medicine composition acting on the intersection targets of hair loss in Experimental Example 2 of the present invention; Figure 4 It is the drug-compound-target-disease network diagram in Experimental Example 3 of the present invention; Figure 5 It is the bar chart of compound connectivity ranking in Experimental Example 3 of the present invention; Figure 6 It is the mulberry diagram of the drug source of TOP compounds in Experimental Example 3 of the present invention; Figure 7 It is the STRING interaction diagram in Experimental Example 4 of the present invention; Figure 8 It is the PPI interaction network diagram in Experimental Example 4 of the present invention; Figure 9 It is the bar chart of core target connectivity ranking in Experimental Example 4 of the present invention; Figure 10 It is the bar chart of GO enrichment analysis in Experimental Example 5 of the present invention; Figure 11 It is the pie chart of GO enrichment analysis in Experimental Example 5 of the present invention; Figure 12 It is the bubble chart of KEGG enrichment analysis in Experimental Example 5 of the present invention; Figure 13 It is the pie chart of KEGG enrichment analysis in Experimental Example 5 of the present invention; Figure 14 It is the mulberry diagram of TOP targets and TOP pathways in Experimental Example 5 of the present invention; Figure 15This is the compound-target-pathway-disease network diagram in Experimental Example 6 of the present invention; Figure 16 This is the docking energy heat map of the Top active ingredients and Top targets in Experimental Example 7 of the present invention; Figure 17 This is the docking diagram of the Top6 compounds and targets in Experimental Example 7 of the present invention; Figure 18 This is the growth condition of the back hair of each group of mice photographed on the 0th, 5th, and 10th days in Experimental Example 8 of the present invention; Figure 19 This is the H&E staining result of the skin tissue in the depilated area on the back of the mice in Experimental Example 8 of the present invention; Figure 20 This is the statistical chart of the hair follicle number in the skin tissue on the back of the mice in Experimental Example 8 of the present invention; Figure 21 This is the Ki67 staining result of the skin tissue in the depilated area on the back of the mice in Experimental Example 8 of the present invention; Figure 22 This is the immunofluorescence staining result of the skin tissue in the depilated area on the back of the mice in Experimental Example 8 of the present invention. Detailed implementation manners

[0020] The present invention will be further described below in conjunction with the accompanying drawings and detailed implementation manners.

[0021] Example: Example 1: Astragalus membranaceus 10g, Panax ginseng 100g, Ligustrum lucidum 100g, Drynaria fortunei 100g, Evodia rutaecarpa 3g, Salvia miltiorrhiza 100g, Polygonum multiflorum 5g, Angelica sinensis 5g, Rehmannia glutinosa var. recens 5g, Rehmannia glutinosa 5g, Coix lacryma-jobi 3g, Poria cocos 5g, and Atractylodes macrocephala 3g.

[0022] Example 2: Astragalus membranaceus 50g, Panax ginseng 200g, Ligustrum lucidum 200g, Drynaria fortunei 200g, Evodia rutaecarpa 50g, Salvia miltiorrhiza 200g, Polygonum multiflorum 50g, Angelica sinensis 50g, Rehmannia glutinosa var. recens 50g, Rehmannia glutinosa 0g, Coix lacryma-jobi 50g, Poria cocos 50g, and Atractylodes macrocephala 50g.

[0023] Example 3: Astragalus membranaceus 10g, Panax ginseng 158g, Ligustrum lucidum 158g, Drynaria fortunei 158g, Evodia rutaecarpa 9g, Salvia miltiorrhiza 158g, Polygonum multiflorum 15g, Angelica sinensis 15g, Rehmannia glutinosa var. recens 15g, Rehmannia glutinosa 12g, Coix lacryma-jobi 6g, Poria cocos 15g, and Atractylodes macrocephala 30g.

[0024] The dosage forms of the hair-nourishing traditional Chinese medicine composition of the present invention include decoctions, powders or ointments prepared from the raw materials in the above weight ratio.

[0025] The following animal experiments use the decoction prepared in Example 3.

[0026] Experimental Example 1: Collection of Active Ingredients of Hair-Nourishing Traditional Chinese Medicine Composition and Target Prediction Using TCMSP / BATMAN / SYMMAP / HERB, collect the chemical components of Astragalus membranaceus, Panax ginseng, Ligustrum lucidum, Drynaria fortunei, Evodia rutaecarpa, Salvia miltiorrhiza, Polygonum multiflorum, Angelica sinensis, Rehmannia glutinosa, Rehmannia glutinosa preparata, Coix lacryma-jobi, Poria cocos, and Atractylodes macrocephala, respectively. Further screen the active ingredients of the hair-nourishing traditional Chinese medicine composition with the screening conditions of oral bioavailability (OB) ≥ 30% and druglikeness (DL) ≥ 0.18. Standardize the gene names of the drug targets obtained from the database using the Uniprot database and remove duplicates. Finally, obtain the active ingredients and action targets of the drug.

[0027] Through the drug database (see the above table), retrieve the components of the hair-nourishing traditional Chinese medicine composition, and screen the active ingredients that meet the conditions of OB ≥ 30% and DL ≥ 0.18. Respectively, 37 active ingredients of Astragalus membranaceus, 83 active ingredients of Panax ginseng, 19 active ingredients of Ligustrum lucidum, 26 active ingredients of Drynaria fortunei, 30 active ingredients of Evodia rutaecarpa, 87 active ingredients of Salvia miltiorrhiza, 15 active ingredients of Polygonum multiflorum, 24 active ingredients of Angelica sinensis, 1 active ingredient of Rehmannia glutinosa, 6 active ingredients of Rehmannia glutinosa preparata, 17 active ingredients of Coix lacryma-jobi, 31 active ingredients of Poria cocos, and 14 active ingredients of Atractylodes macrocephala are obtained, totaling 282 active ingredients, see Figure 1 . Retrieve the potential action targets of the drug active ingredients through the database. After deleting duplicate targets, a total of 2214 drug targets are obtained, see Figure 2 .

[0028] Experimental Example 2: Collection of Related Targets of Alopecia and Screening of Therapeutic Action Targets of Hair-Nourishing Traditional Chinese Medicine Composition Retrieve the disease-related targets from the OpenTargets / OMIM / GeneCards database. Use "Alopecia" as the keyword to retrieve the disease-related targets, summarize and standardize the gene names of the disease targets, and remove duplicates. Take the intersection genes of alopecia as the final genes of the disease. Finally, obtain the potential targets of alopecia. And draw a Venn diagram of the target intersection situation between different databases. Take the intersection of the compound targets obtained in Experimental Example 1 and the disease targets downloaded from the database in R to obtain the common targets as the targets related to the treatment of alopecia by the hair-nourishing traditional Chinese medicine composition for the next step of analysis. Use the R4.2.1 VennDiagram 1.7.3 and UpSetR 1.4.0 packages to draw the Venn diagram of the target intersection situation between different databases respectively.

[0029] The hair loss disease retrieval was screened according to the threshold from the retrieval results of the OpenTargets Score>=0.1 / OMIM / GeneCards Score>=5 database. The gene names of the standardized disease targets were summarized, duplicates were removed, and the intersection was taken to obtain 410 disease targets. The intersection was taken with the active ingredient targets of the hair-nourishing traditional Chinese medicine composition. After mapping the two, 113 common action targets were determined as the potential targets of the hair-nourishing traditional Chinese medicine composition acting on hair loss, as shown in Figure 3 。

[0030] Experimental Example 3: Construction and Analysis of Drug-Compound-Target-Disease Network The one-to-one correspondence between the active ingredients of the hair-nourishing traditional Chinese medicine composition and the targets was imported into the Cytoscape 3.9.1 software to construct a drug-compound-target-disease network diagram. The nodes represent compounds and targets respectively, and the edges represent the interaction relationship between compounds and targets. The Network Analyzer plugin was used to analyze the network topology properties and calculate the degree value of the nodes (degree).

[0031] The drug-compound-target-disease network is shown in Figure 4 . This network diagram has 377 nodes and 1492 edges. The edges represent the targeting relationship between compounds and targets. Among them, 250 active ingredients act on 113 targets (some compound targets not in the intersection were deleted). The Network Analyzer plugin was used to analyze the network topology properties. In the network diagram, the size of the node represents the size of the degree value of the node. The larger the degree value, the stronger the hub role of the node in this network. The degree value ranking of the top 20 main components Figure 5 . Based on this analysis, it was found that the main active ingredients in the active ingredients include quercetin, wogonin, baicalin, berberine, luteolin, beta-sitosterol, Delta-D, etc. Different shapes in the figure represent drugs, compounds and targets, and the depth of color represents the size of the degree weight in the network. The Sankey diagram of the drug source of the TOP compound, see Figure 6 。

[0032] Experimental Example 4: Construction and Analysis of Protein-Protein Interaction Network Construct a protein - protein interaction (PPI) network diagram using the STRING database. Import the intersection targets related to hair loss treated by the hair - nourishing traditional Chinese medicine composition into the STRING database. Select the species "Homo sapiens" and set the confidence level to ≥0.4 to obtain the interaction relationships between proteins, and save the TSV and PNG files. Import them into the Cytoscape software, delete the isolated targets without intersections, analyze the network topology properties, and screen the core targets according to the degree value sorting of the topological structure (CC / BC / degree sorting) / Use the MCC algorithm of the cytohubba plugin to screen the core targets.

[0033] Import 113 intersection targets related to diseases treated by drugs into the STRING database to obtain the PPI relationship, and then import it into the Cytoscape software for analysis, as shown in Figure 7 . This network diagram has 172 nodes and 768 edges. The edges represent the association relationships between the targets (delete the isolated targets without intersections). Use the Network Analyzer plugin to analyze the network topology properties, calculate the degree values, and set the color depth and node size to reflect the degree value size. Select the top 30 core nodes according to the degree value (CC / BC / degree) ranking and draw the network diagram, as shown in Figure 8 . The core targets are TNF, IL6, IL1B, CTNNB1, BCL2, TGFB1, IFNG, IL10, IL2, etc., which are used as the key targets for the hair - nourishing traditional Chinese medicine composition to treat hair loss, as shown in Figure 9 .

[0034] Experimental Example 5: GO functional enrichment and KEGG pathway enrichment analysis Use the R clusterProfiler4.6.2 package to perform gene ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the hair - nourishing traditional Chinese medicine composition and the related targets of hair loss respectively. For the GO enrichment analysis, use P < 0.05 as the screening condition, and select the top 20 biological terms with P - value less than 0.05 in the results to draw a bar chart. For the KEGG enrichment analysis, also use P < 0.05 as the significant enrichment screening condition, and select the top 30 signaling pathways with P < 0.05 to draw a bubble chart for visual analysis.

[0035] Based on R clusterProfiler, GO enrichment analysis was performed on 113 intersection targets related to diseases treated with drugs. The top 15 entries of biological process (BP), molecular function (MF), and cellular component (CC) with P < 0.05 were selected respectively to draw bar charts, as shown in Figure 10 , Figure 11. The vertical axis represents each entry of GO, and the horizontal axis represents the number of targets enriched to that entry. BP mainly involves positive regulation of lymphocyte activation, positive regulation of leukocyte activation, positive regulation of lymphocyte differentiation, positive regulation of cell activation, positive regulation of leukocyte differentiation, positive regulation of cell-cell adhesion, T cell differentiation, regulation of T cell activation, positive regulation of cytokine production, monocyte differentiation, positive regulation of leukocyte-cell, α-β T cell activation, etc.; MF mainly includes cytokine receptor binding cytokine activity, receptor ligand activity, signal receptor activator activity, steroid binding, growth factor receptor binding, growth hormone receptor binding, non-transmembrane protein tyrosine kinase, growth factor activity, nuclear receptor activity, ligand-activated transcription factor activity, hormone receptor binding, 3',5'-cyclic adenosine monophosphate phosphodiesterase activity, etc. CC mainly involves the outer lateral surface of the plasma membrane, membrane microdomains, R polymerase II transcription regulatory factors, transcription regulatory complexes, focal adhesions, endolysosomal membranes, cell-substrate junctions, hemidesmosomes, lateral plasma membranes, endolysosomes, lumens of platelet α granules, growth factor complexes, etc. KEGG pathway enrichment analysis was performed on the targets, and a total of 142 signaling pathways were enriched. In this invention, the top 30 pathways with P < 0.05 in the downloaded results were selected to draw a bubble chart for visual analysis, as shown in Figure 12 , Figure 13. The enrichment results show that it mainly involves signaling pathways such as inflammatory bowel disease, Th17 cell differentiation, JAK-STAT signaling pathway, graft-versus-host disease, influenza A, allograft rejection, cytokine-cytokine receptor interaction, type I diabetes, rheumatoid arthritis, hepatitis B, AGE-RAGE signaling pathway in diabetes, proteoglycans in cancer, Chagas disease, etc. These pathways communicate with each other and play an important role in the occurrence and development of diseases. In addition, this invention also drew a Sankey diagram of the TOP pathways related to the TOP targets, as shown in Figure 14 .

[0036] Experimental Example 6, Construction and Analysis of Drug-Compound-Target-Pathway-Disease Network To discover the potential mechanisms of drug action, the core TOP pathways, key targets, and active ingredients therein were selected to construct a "drug-compound-target-pathway-disease" network. To better demonstrate the interconnections and synergistic effects of the important active ingredients, targets, and the above-mentioned important pathways in the hair-nourishing traditional Chinese medicine composition.

[0037] As Figure 15 shown, this network consists of 407 nodes and 1807 edges (250 components, 113 targets, 30 signaling pathways, drugs, and diseases). Different node shapes represent different active ingredients, key targets, and regulatory pathways in the hair-nourishing traditional Chinese medicine composition, and the color depth represents the degree weight size in the network. Each active ingredient corresponds to multiple targets, and each target connects multiple components, reflecting the mechanism of action of the multi-component and multi-target prevention and treatment of hair loss in the hair-nourishing traditional Chinese medicine composition.

[0038] Experimental Example 7, Molecular Docking The important compounds screened out were docked with the core target molecules. The 3D structures of the main active compounds in the hair-nourishing traditional Chinese medicine composition were downloaded from the PubChem database, and the 3D structures of the core target proteins were downloaded from the Protein Data Bank (https: / / www.rcsb.org / ) database. The core target proteins were preliminarily processed, such as removing solvent molecules, using the Pymol 2.6.0 software, and then further hydrogenated and charged using AutoDock Tools 1.5.7. The core target proteins and active compounds were saved as files in the "pdbqt" format, and the appropriate grid positions and sizes were set. Finally, the docking of the ingredients and targets was completed by Autodock Vina. The docking results were visualized using the PyMOL software.

[0039] Combining the ranking of the active ingredient degree values in the active ingredient-target network diagram and literature research, the present invention performed molecular docking on the active ingredients MOL000006, MOL000098, MOL000173, MOL000358, MOL001454, MOL001638, MOL002776, MOL003187, MOL008691, MOL010861 in the hair-nourishing traditional Chinese medicine composition and the top 10 core targets BCL2, CTNNB1, IFNG, IL10, IL1B, IL2, IL4, IL6, TGFB1, TNF in the PPI network. The docking results are shown in Figure 16The smaller the binding energy, the higher the binding activity, and the easier it is for the compound to bind to the target. Binding energy ≤-5.0 kcal / mol indicates good binding, and binding energy ≤-7.5 kcal / mol indicates very good binding. Molecular docking was visualized using PyMOL 2.6.0 software and Discovery Studio. The 3D and 2D images of the top 6 compounds docking with the target are shown in Figure 17 .

[0040] Experimental Example 8: Animal Experiment A total of 20 C57 mice were purchased from the Animal Medical Center of Yangzhou University. After each group of mice was anesthetized by intraperitoneal injection of 7.5% chloral hydrate solution, a small electric shaver was used to carefully shave the hair on the back of the mice. The shaved area was about 3cm×4cm. According to the instructions for use of the depilatory cream, apply the depilatory cream to the skin area where the hair has been shaved, wait for 10 minutes after application, and then use a cotton ball or gauze soaked in physiological saline to wipe off the depilatory cream and residual hair. At this time, it can be observed that the skin on the back of the mouse is light pink. It is divided into two experiments, 10 hair care prescriptions and 10 blank groups. The decoction of the hair care Chinese medicine composition in Example 3 was applied to the hair removal area of ​​the hair care prescription group every day, and photos were taken on the 0th, 5th, and 10th days. After taking photos on the 10th day, the mice were killed for sampling.

[0041] Result 1: Hair growth After the hair removal treatment, starting from the first day after hair removal, the hair removal areas on the back of each group of mice were subjected to corresponding experimental interventions at 10 am every day, and the hair growth was observed. On the 0th, 5th, and 10th days after hair removal, the hair removal areas on the back of the mice were photographed. On the day of hair removal, the hair removal areas on the back of each group of mice were pink or light gray, and no hair grew; on the 5th day after hair removal, it can be observed that the mice in the hair-nourishing prescription group had more hair growth, while only some areas of the blank group had less hair growth; on the 10th day, the hair of the mice in the hair-nourishing prescription group had basically grown out, and the new hair was shiny and black in color; the blank group mice had less hair growth, or even only light gray hair, and the skin edges of the new and old hair had obvious boundary lines, and the hair at the hair removal boundary grew slowly. The blank group and the normal saline group had terminal hair on the back of the mice, but the growth was not as good as the hair-nourishing prescription group ( Figure 18 ).

[0042] Result 2: HE pathological staining On the 10th day of the experimental treatment, the mouse skin was taken for pathological sections and H&E staining to observe the hair growth of the mice. It can be seen that the number of hair follicles in the blank group is relatively small. The horizontal comparison shows that the growth of hair follicles in the hair nourishing prescription group is more than that in the blank group ( Figure 19 ). Statistics and comparison of the number of hair follicles in each group under a microscope (×20) ( Figure 20 ).

[0043] Result 3: Immunohistochemical staining Immunohistochemical staining with the cell proliferation index protein Ki67 was performed on paraffin sections of mouse dorsal skin tissue. The positive staining result was brown-yellow granules, and the darker the color, the more obvious it was, indicating more expression of Ki67 protein, that is, the more vigorous the cell proliferation and division. The results showed Figure 21 , the expression of KI67 protein in hair follicle cells of the hair-nourishing formula group was significantly higher than that of the blank group, suggesting that the hair-nourishing traditional Chinese medicine composition promoted the proliferation activity of hair follicle cells after treatment.

[0044] Result 4: Immunofluorescence staining After paraffin sections were made from skin tissue specimens of mice in each group, immunofluorescence staining was performed. The marker was the cell proliferation protein Ki67 (red), the cell nucleus was labeled with DAPI (blue), and apoptotic cells were stained with Tunel (green). The positive result of DAPI staining was blue, the positive result of Ki67 staining was red, and the positive result of Tunel staining was green. The darker the red of Ki67 staining, the more obvious it was, indicating more expression of Ki67 protein, that is, the more vigorous the cell proliferation and division; the darker the green of Tunel staining, the more apoptosis. The results of immunofluorescence staining were consistent with those of immunohistochemistry ( Figure 22 ), the expression of KI67 protein in hair follicle cells of the hair-nourishing formula group was significantly higher than that of the blank group, and there were basically no apoptotic cells in the hair-nourishing formula group, further confirming that the hair-nourishing traditional Chinese medicine composition promoted the proliferation of hair follicle cells after treatment.

Claims

1. A Chinese medicinal composition for hair care for preventing and treating hair loss, characterized in that: The invention is prepared from the following raw materials in proportions by weight: 10-50 parts of astragalus, 100-200 parts of ginseng, 100-200 parts of glossy privet, 100-200 parts of drynaria, 3-50 parts of evodia, 100-200 parts of salvia, 5-50 parts of polygonum multiflorum, 5-50 parts of angelica, 5-50 parts of raw rehmannia, 5-50 parts of prepared rehmannia, 3-50 parts of coix seeds, 5-50 parts of tuckahoe and 3-50 parts of atractylodes.

2. The hair nourishing Chinese medicine composition for preventing and treating hair loss according to claim 1, characterized in that: The weight ratio of the raw materials is: 10 parts of astragalus, 158 parts of ginseng, 158 parts of Ligustrum lucidum, 158 parts of drynaria, 9 parts of Evodia rutaecarpa, 158 parts of Salvia miltiorrhiza, 15 parts of Polygonum multiflorum, 15 parts of Angelica sinensis, 15 parts of Rehmannia root, 12 parts of Rehmannia root, 6 parts of Coix seed, 15 parts of Poria and 30 parts of Atractylodes macrocephala.

3. The hair nourishing Chinese medicinal composition for preventing and treating hair loss as claimed in claim 1 or 2, characterized in that: It can be made into decoction, powder or paste.

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