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

Abstract

The hair-growing traditional Chinese medicine composition is prepared from the following raw materials: herba houttuyniae, radix scutellariae, cacumen biotae, radix sophorae flavescentis, rhizoma cimicifugae, perilla frutescens, cortex dictamni, lalang grass rhizome, fructus kochiae, rhizoma acori calami, radix bupleuri, radix pulsatillae and frankincense. Based on the principle of clearing heat to cool blood and dispelling wind and promoting diuresis, the hair-growing traditional Chinese medicine composition prepared from herba houttuyniae, radix scutellariae, cacumen biotae, radix sophorae flavescentis, rhizoma cimicifugae, perilla frutescens, cortex dictamni, lalang grass rhizome, fructus kochiae, rhizoma acori calami, radix bupleuri, radix pulsatillae and frankincense contains multiple active ingredients and can act on multiple key targets related to alopecia; further, the action mechanism for treating diseases can be synergistically exerted by participating in regulation and control of multiple pathways, such as pathways.

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 is a common hair problem. Approximately 50% of men and 25% of women worldwide will experience hair loss at some stage. In recent years, the problem of hair loss has shown a trend of getting younger. Among the post-90s generation, more than 50% of people believe that staying up late and excessive stress are the main causes of their hair loss. The causes of hair loss are very complex, including genetic factors, hormone levels, nutritional intake, life stress, environmental pollution, and drug side effects, etc. Traditional Chinese medicine believes that hair loss is closely related to the functions of the zang-fu organs and qi and blood in the human body. Common syndromes include damp-heat accumulation, blood heat running amok, and heat-toxin disturbing upward.

[0003] The clinical application of existing hair loss prevention and treatment products has the following limitations: Topical medications represented by minoxidil need to be continuously applied externally, which may induce local adverse reactions such as contact dermatitis or hirsutism; Although finasteride preparations can inhibit the production of dihydrotestosterone (DHT), they have gender applicability limitations (only for male androgenetic alopecia), and may cause systemic side effects such as endocrine disorders and sexual dysfunction. Their action targets are single and it is difficult to cover the complex pathological mechanisms of hair loss. Anti-hair loss and hair care cleaning products (containing active ingredients such as ketoconazole / caffeine) mainly reflect local cleaning effects. Although they can achieve the short-term goal of oil control, they cannot intervene in the imbalance of the internal microenvironment due to limited transdermal absorption efficiency, and long-term use may instead interfere with the normal flora homeostasis of the scalp. Although traditional Chinese medicine compound preparations have the advantage of overall regulation, they generally have two deficiencies: First, the clinical action mechanism lacks the support of modern pharmacology evidence; Second, the formula compatibility mostly focuses on a single treatment principle (such as cooling blood or removing dampness), and it is difficult to achieve multi-dimensional coordinated regulation (such as simultaneously taking into account compound functions such as clearing heat and detoxifying, cooling blood and stopping bleeding), affecting the systematicness of the treatment effect. 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 ideas and theoretical basis for health preservation and care for hair loss.

[0005] In order to achieve the above purpose, the technical scheme adopted by the present invention is as follows: A hair-nourishing traditional Chinese medicine composition for preventing and treating hair loss is made from raw materials with the following weight part ratios: 5 - 20 parts of Houttuynia cordata, 5 - 20 parts of Scutellaria baicalensis, 80 - 150 parts of Platycladus orientalis, 5 - 20 parts of Sophora flavescens, 1 - 10 parts of Cimicifuga foetida, 5 - 20 parts of Perilla frutescens, 5 - 20 parts of Dictamnus dasycarpus, 3 - 20 parts of Imperata cylindrica, 5 - 20 parts of Kochia scoparia, 3 - 20 parts of Acorus calamus, 1 - 10 parts of Bupleurum chinense, 1 - 10 parts of Pulsatilla chinensis, and 1 - 10 parts of Olibanum.

[0006] The preferred weight ratio of each raw material is as follows: 9 parts of Houttuynia cordata, 9 parts of Scutellaria baicalensis, 128 parts of Platycladus orientalis leaf, 12 parts of Sophora flavescens, 3 parts of Cimicifuga foetida, 9 parts of Perilla frutescens, 12 parts of Dictamnus dasycarpus, 6 parts of Imperata cylindrica, 9 parts of Kochia scoparia, 6 parts of Acorus calamus, 3 parts of Bupleurum chinense, 3 parts of Pulsatilla chinensis, and 3 parts of Olibanum.

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

[0008] The beneficial effects of the present invention compared with the prior art are as follows: Traditional Chinese medicine has a long history in preventing and treating hair loss and is of great significance in traditional medicine. Traditional Chinese medicines for preventing and treating hair loss mainly include those for tonifying deficiency, clearing heat, and promoting blood circulation to remove blood stasis. In addition to having the pharmacological activities of hair growth and hair nourishment, such traditional Chinese medicines also have the characteristics of regulating abnormal hair follicle growth through multiple pathways, multiple targets, and multiple channels. The present invention will conduct network pharmacology and molecular docking on traditional Chinese medicines for tonifying deficiency, clearing heat, promoting blood circulation to remove blood stasis, and their active ingredients and the target proteins for anti-hair loss and hair growth effects, screen the key compounds that play roles, and verify the effectiveness of the traditional Chinese medicine composition of the present invention by combining animal experiments, which is expected to provide new growth momentum for the relevant product market and provide a theoretical basis and new ideas for the prevention and treatment of hair loss.

[0009] Platycladus orientalis leaf is the monarch drug of the traditional Chinese medicine composition of the present invention and has the effects of cooling blood to stop bleeding and clearing heat and detoxifying. It is the main drug of the traditional Chinese medicine composition of the present invention. Hair loss is often related to blood heat running wild and heat toxin disturbing upward. Platycladus orientalis leaf can effectively reduce hair follicle bleeding caused by blood heat through cooling blood to stop bleeding, stabilize the foundation of hair follicles, and prevent hair loss; its function of clearing heat and detoxifying can also remove the heat toxin on the scalp, improve the scalp environment, and create good conditions for hair growth.

[0010] Scutellaria baicalensis, Sophora flavescens, Dictamnus dasycarpus, and Kochia scoparia are the ministerial herbs in the traditional Chinese medicine composition of the present invention. Scutellaria baicalensis is cold in nature and bitter in taste, and has the effects of clearing heat and drying dampness, cooling blood and detoxifying. Hair loss is mostly due to exuberant liver fire and internal accumulation of dampness and heat. By clearing heat and drying dampness, Scutellaria baicalensis can effectively inhibit the flaring up of liver fire and reduce the invasion of dampness and heat on hair follicles. At the same time, its effect of cooling blood and detoxifying complements that of Platycladus orientalis, jointly clearing away heat toxin on the scalp and promoting the healthy growth of hair; Sophora flavescens is bitter in taste and cold in nature, and has the effects of clearing heat and drying dampness, killing insects and promoting diuresis. Hair loss patients often have symptoms such as greasy scalp and itching. By clearing heat and drying dampness, Sophora flavescens can effectively reduce the secretion of scalp oil and relieve scalp itching. At the same time, its insecticidal effect can also kill parasites on the scalp and prevent hair loss caused by insect pests; Dictamnus dasycarpus is cold in nature and bitter in taste, and has the effects of clearing heat and drying dampness, dispelling wind and detoxifying. Hair loss is related to the invasion of wind pathogen and internal accumulation of dampness and heat. By dispelling wind and detoxifying, Dictamnus dasycarpus can effectively disperse wind pathogen on the scalp and relieve symptoms such as scalp itching and swelling caused by wind pathogen. At the same time, its effect of clearing heat and drying dampness can also further improve the scalp environment and promote hair growth; Kochia scoparia is cold in nature and bitter in taste, and has the effects of clearing heat and promoting diuresis, dispelling wind and relieving itching. Hair loss patients often have scalp itching and swelling due to the accumulation of dampness and heat and the invasion of wind pathogen. By clearing heat and promoting diuresis, Kochia scoparia can effectively remove dampness and heat on the scalp and relieve itching symptoms. At the same time, its effect of dispelling wind can also disperse wind pathogen, stabilize hair follicles and prevent hair loss.

[0011] Houttuynia cordata, Cimicifuga foetida, Perilla frutescens, Bupleurum chinense, and Pulsatilla chinensis are the adjuvant herbs in the traditional Chinese medicine composition of the present invention. Houttuynia cordata is slightly cold in nature and pungent in taste, and has the effects of clearing heat and detoxifying, expelling pus and detumescence. Hair loss is often due to internal accumulation of heat toxin and poor qi and blood circulation. By clearing heat and detoxifying, Houttuynia cordata can effectively remove heat toxin on the scalp, promote qi and blood circulation, and provide good blood circulation support for hair growth; Cimicifuga foetida is cold in nature and bitter, pungent in taste, and has the effects of clearing heat and detoxifying, lifting yang qi. Hair loss is related to insufficient yang qi and failure of qi and blood to ascend. By lifting yang qi, Cimicifuga foetida can effectively improve the qi and blood circulation of the scalp, improve the nutrient supply of hair follicles, and promote hair growth; Perilla frutescens is warm in nature and pungent in taste, and has the effects of relieving exterior syndrome and dispelling cold, promoting qi movement and regulating the stomach. Hair loss patients often have exterior contraction of wind-cold and poor qi and blood circulation. By relieving exterior syndrome and dispelling cold, Perilla frutescens can effectively disperse wind-cold pathogen on the scalp and relieve symptoms such as scalp tightness and pain caused by wind-cold. At the same time, its effect of promoting qi movement can also promote qi and blood circulation and provide power for hair growth; Bupleurum chinense is cool in nature and bitter, pungent in taste, and has the effects of soothing the liver and relieving depression, lifting yang qi. Hair loss is related to stagnation of liver qi and poor qi and blood circulation. By soothing the liver and relieving depression, Bupleurum chinense can effectively relieve symptoms such as chest distress and emotional discomfort caused by stagnation of liver qi. At the same time, its effect of lifting yang qi can also improve the qi and blood circulation of the scalp and promote hair growth; Pulsatilla chinensis is cold in nature and bitter in taste, and has the effects of clearing heat and detoxifying, cooling blood and stopping dysentery. Hair loss is often due to internal accumulation of heat toxin and reckless flow of blood heat. By clearing heat and detoxifying, Pulsatilla chinensis can effectively remove heat toxin on the scalp, cool blood and stop bleeding, stabilize the root of hair follicles, and prevent hair loss.

[0012] Frankincense, Imperata cylindrica, and Acorus calamus are the envoy drugs of the traditional Chinese medicine composition of the present invention. Frankincense is warm in nature and pungent in taste, and has the functions of promoting blood circulation and qi movement, and relieving pain. Hair loss is related to poor qi and blood circulation and meridian blockage. Frankincense can effectively promote blood circulation in the scalp through promoting blood circulation and qi movement, relieve symptoms such as scalp pain and tightness caused by poor qi and blood circulation, and at the same time its pain-relieving effect can also reduce the discomfort caused by hair loss; Imperata cylindrica is cold in nature and sweet in taste, and has the functions of clearing heat and cooling blood, and promoting diuresis and relieving stranguria. Hair loss is often caused by blood heat running wild and heat toxin accumulating internally. Imperata cylindrica can effectively reduce blood heat through clearing heat and cooling blood, prevent hair follicle bleeding caused by blood heat running wild, and at the same time its diuretic effect can also help discharge heat toxin in the body and improve the scalp environment; Acorus calamus is warm in nature and pungent, bitter in taste, and has the functions of opening the orifices and waking up the mind, and resolving dampness and regulating the stomach. Hair loss is related to internal obstruction of damp turbidity and poor qi and blood circulation. Acorus calamus can effectively resolve damp turbidity on the scalp through resolving dampness and regulating the stomach, improve the scalp environment, and at the same time its effect of opening the orifices and waking up the mind can also promote qi and blood circulation and provide power for hair growth.

[0013] The traditional Chinese medicine composition of the present invention takes Platycladus orientalis as the main drug, supplemented by Scutellaria baicalensis, Sophora flavescens, Dictamnus dasycarpus, Kochia scoparia and other drugs for clearing heat and detoxifying, dispelling wind and drying dampness, jointly clearing heat toxin and damp heat in the scalp, and improving the scalp environment; assisted by Houttuynia cordata, Cimicifuga foetida, Perilla frutescens, Bupleurum chinense, Pulsatilla chinensis and other drugs for soothing the liver and relieving depression, lifting yang qi, promoting qi and blood circulation, promoting qi and blood circulation, and stabilizing the hair follicle foundation; made with frankincense, Imperata cylindrica, Acorus calamus and other drugs for promoting blood circulation and qi movement, clearing heat and cooling blood, resolving dampness and regulating the stomach, further improving the scalp environment, promoting hair growth, and the whole formula together exerts the effects of clearing heat and detoxifying, dispelling wind and drying dampness, soothing the liver and relieving depression, lifting yang qi, promoting qi and blood circulation, and stabilizing the hair follicle, is applicable to various types of hair loss problems, can effectively promote hair growth, and improve hair loss symptoms.

[0014] Through the TCMSP / BATMAN / HERB / ETCM traditional Chinese medicine databases and analysis platforms, the active ingredients and potential action targets of each medicinal flavor in the traditional Chinese medicine composition proposed in the present invention were screened and summarized. The disease-related target summaries were retrieved in the DisGeNET / CTD / DrugBank databases, and a compound target-disease Venn diagram was constructed to obtain the intersection target list between the drug targets and the disease targets; the Cytoscape 3.9.1 software was used to draw the drug-component-target-disease network diagram. The STRING database and Cytoscape were used to construct a protein-protein interaction PPI network diagram to screen the core targets among the intersection targets. The clusterProfiler package of the R 4.2.1 software was used 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-growing traditional Chinese medicine composition in the present invention on hair loss, and a drug-component-target-pathway network diagram was constructed with Cytoscape; finally, AutoDock Tools was used to perform molecular docking on the key active ingredients and core targets to verify the results of network pharmacology analysis.

[0015] 188 active ingredients, 1349 active ingredient targets, and 523 hair loss-related targets were screened out from the hair-growing formula through the oral bioavailability OB and the drug-likeness parameter DL; 195 intersection targets; 2003 biological process BP entries, 39 cellular component CC entries, and 126 molecular function MF entries (p.adjust < 0.01) were obtained through GO function enrichment analysis; 174 (P < 0.05) significant signaling pathways were screened out through KEGG pathway enrichment analysis, mainly involving Lipid and atherosclerosis, AGE-RAGE signaling pathway in diabetic, Platinum drug resistance, Chemical carcinogenesis - receptor activation, Multiple signaling pathways such as Apoptosis, Hepatitis B, Prostate cancer, Measles, Kaposisarcoma-associated herpesvirus infection, TNF signaling pathway, and Endocrine resistance; molecular docking showed that the TOP active ingredient MOL002776 screened from the drug had a good affinity with the screened core target CYP1A1. MOL000173, MOL002714, MOL002773, MOL002776, and MOL008691 all had good affinity with the screened core target CYP3A4.

[0016] Based on the principle of "clearing heat and cooling blood, expelling wind and removing dampness", the hair-growing traditional Chinese medicine composition of Houttuynia cordata, Scutellaria baicalensis, Platycladus orientalis, Sophora flavescens, Cimicifuga foetida, Perilla frutescens, Dictamnus dasycarpus, Imperata cylindrica, Kochia scoparia, Acorus calamus, Bupleurum chinense, Pulsatilla chinensis, and Olibanum contains a variety of active ingredients, which can act on multiple key targets related to hair loss, and may then synergistically play a disease treatment mechanism by participating in the regulation of multiple pathways such as the [pathways]. The formula follows the compatibility of "sovereign, ministerial, assistant, and courier": the sovereign drug Platycladus orientalis exerts the effect of cooling blood and detoxifying, and stabilizes the hair follicle foundation for blood-heat type hair loss; the ministerial drugs Scutellaria baicalensis and Sophora flavescens cooperate to clear heat and dry dampness, and improve the scalp greasiness and micro-inflammatory environment caused by the accumulation of dampness and heat; the assistant drugs Bupleurum chinense and Cimicifuga foetida regulate the qi and blood circulation by sooth the liver and ascending yang, and promote the nutrient supply of hair follicles; the courier drugs Olibanum and Acorus calamus activate blood circulation and resolve dampness, and maintain the scalp metabolic homeostasis. Animal experiments verified that it significantly improved hair loss symptoms and played a role by regulating the apoptosis pathway. Compared with existing therapies, this compound breaks through the limitation of a single mechanism: it avoids the risk of local irritation compared with minoxidil, expands gender applicability compared with finasteride, and improves the scientificity of the formula compared with traditional Chinese medicine. The research provides new ideas for the interpretation of the pathological mechanism of hair loss and the development of modern traditional Chinese medicine products. Brief Description of the Drawings

[0017] Figure 1 It is the intersection diagram of active ingredients among different databases in Experimental Example 1 of the present invention; Figure 2 It is the intersection diagram of targets among different databases in Experimental Example 1 of the present invention; Figure 3 It is the Venn diagram of the hair-growing 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 the compound connectivity ranking in Experimental Example 3 of the present invention; Figure 6It is the Sankey diagram of the drug source of the TOP compound 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 the 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 Sankey diagram of the TOP target - TOP pathway in Experimental Example 5 of the present invention; Figure 15 It is the compound - target - pathway - disease network diagram in Experimental Example 6 of the present invention; Figure 16 It is the heat map of the docking energy between the Top active ingredients and Top targets in Experimental Example 7 of the present invention; Figure 17 It is the docking diagram of the Top6 compounds and targets in Experimental Example 7 of the present invention; Figure 18 It is the growth condition of the back hair of mice in each group photographed on the 0th, 5th, and 10th days of the experiment in Experimental Example 8 of the present invention; Figure 19 It is the H&E staining result of the skin tissue of the depilated area on the back of mice in Experimental Example 8 of the present invention; Figure 20 It is the statistical chart of the hair follicle number of the skin tissue on the back of mice in Experimental Example 8 of the present invention; Figure 21 It is the Ki67 staining result of the skin tissue of the depilated area on the back of mice in Experimental Example 8 of the present invention; Figure 22 It is the immunofluorescence staining result of the skin tissue of the depilated area on the back of mice in Experimental Example 8 of the present invention. Detailed implementation manners

[0018] The present invention will be further described below in conjunction with the accompanying drawings and detailed implementation manners.

[0019] Examples: Example 1: 5 g of Houttuynia cordata, 5 g of Scutellaria baicalensis, 80 g of Platycladus orientalis, 5 g of Sophora flavescens, 1 g of Cimicifuga foetida, 5 g of Perilla frutescens, 5 g of Dictamnus dasycarpus, 3 g of Imperata cylindrica, 5 g of Kochia scoparia, 3 g of Acorus calamus, 1 g of Bupleurum chinense, 1 g of Pulsatilla chinensis, and 1 g of Olibanum.

[0020] Example 2: 20 g of Houttuynia cordata, 20 g of Scutellaria baicalensis, 150 g of Platycladus orientalis, 20 g of Sophora flavescens, 10 g of Cimicifuga foetida, 20 g of Perilla frutescens, 20 g of Dictamnus dasycarpus, 20 g of Imperata cylindrica, 20 g of Kochia scoparia, 20 g of Acorus calamus, 10 g of Bupleurum chinense, 10 g of Pulsatilla chinensis, and 10 g of Olibanum.

[0021] Example 3: 9 g of Houttuynia cordata, 9 g of Scutellaria baicalensis, 128 g of Platycladus orientalis, 12 g of Sophora flavescens, 3 g of Cimicifuga foetida, 9 g of Perilla frutescens, 12 g of Dictamnus dasycarpus, 6 g of Imperata cylindrica, 9 g of Kochia scoparia, 6 g of Acorus calamus, 3 g of Bupleurum chinense, 3 g of Pulsatilla chinensis, and 3 g of Olibanum.

[0022] The dosage forms of the hair growth traditional Chinese medicine composition of the present invention include decoction, powder, or ointment prepared from the raw materials in the above weight ratio.

[0023] The following animal experiments use the decoction prepared in Example 3.

[0024] Experimental Example 1: Collection of Active Ingredients in the Hair Growth Traditional Chinese Medicine Composition and Target Prediction Using TCMSP / BATMAN / HERB / ETCM, collect the chemical components of Houttuynia cordata, Scutellaria baicalensis, Platycladus orientalis, Sophora flavescens, Cimicifuga foetida, Perilla frutescens, Dictamnus dasycarpus, Imperata cylindrica, Kochia scoparia, Acorus calamus, Bupleurum chinense, Pulsatilla chinensis, and Olibanum respectively. Further screen out the active ingredients of the hair growth formula with the screening conditions of oral bioavailability (OB) ≥ 30% and drug likeness (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.

[0025] Through the drug database, retrieve the components of the hair growth traditional Chinese medicine composition, and screen out the active ingredients that meet OB ≥ 30% and DL ≥ 0.18. Respectively, 11 active ingredients of Houttuynia cordata, 48 active ingredients of Scutellaria baicalensis, 12 active ingredients of Platycladus orientalis, 40 active ingredients of Sophora flavescens, 19 active ingredients of Cimicifuga foetida, 18 active ingredients of Perilla frutescens, 20 active ingredients of Dictamnus dasycarpus, 9 active ingredients of Imperata cylindrica, 6 active ingredients of Kochia scoparia, 1 active ingredient of Acorus calamus, 24 active ingredients of Bupleurum chinense, 16 active ingredients of Pulsatilla chinensis, and 12 active ingredients of Olibanum are obtained, totaling 188 active ingredients. See Figure 1。The potential action targets of the active ingredients of the drug were obtained through the database. After removing duplicate targets, a total of 1349 drug targets were obtained, as shown in Figure 2 。

[0026] Experimental Example 2: Collection of targets related to alopecia and screening of therapeutic action targets of traditional Chinese medicine compositions for hair growth Retrieve the disease-related targets from the DisGeNET / CTD / DrugBank databases. Use "Alopecia" as the keyword to retrieve the disease-related target summary, standardize the gene names of the disease targets and remove duplicates, and 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 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 traditional Chinese medicine compositions for hair growth, which are used for the next 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 between different databases respectively.

[0027] The retrieval of alopecia diseases was screened according to the thresholds of the retrieval results of DisGeNET Score >= 0.1 / CTD Score >= 60 / DrugBank databases. The gene names of the disease targets were standardized and duplicates were removed, and the intersection was taken to obtain 523 disease targets. The intersection was taken with the active ingredient targets in the traditional Chinese medicine compositions for hair growth. After mapping the two, 195 common action targets were determined as the potential targets of the traditional Chinese medicine compositions for hair growth acting on alopecia, as shown in Figure 3 。

[0028] Experimental Example 3: Construction and analysis of the drug-compound-target-disease network Import the one-to-one correspondence between the active ingredients of the hair growth formula and the targets 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 relationships between compounds and targets. Use the Network Analyzer plugin to analyze the network topology properties and calculate the degree value (degree) of the nodes.

[0029] The drug-compound-target-disease network is shown in Figure 4 , this network diagram has 377 nodes and 2388 edges, and the edges represent the targeting relationships between compounds and targets. Among them, 168 active ingredients act on 195 targets (delete some compound targets that are not in the intersection). Use the Network Analyzer plugin 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 componentsFigure 5 。Based on this analysis, it was found that the main active ingredients in the active ingredient include quercetin, wogonin, alpha-carotene / beta, epsilon-carotene, kaempferol, baicalein, luteolin, etc. Different shapes in the figure represent drugs, compounds and targets, and the depth of color represents the degree weight size in the network. The Sankey diagram of the drug source of the TOP compound is shown in Figure 6 。

[0030] Experimental Example 4: Construction and Analysis of Protein-Protein Interaction Network Use the STRING database to construct a protein-protein interaction (PPI) network diagram. Import the intersection targets related to hair loss of the hair-growing traditional Chinese medicine composition into the STRING database, select "Homo sapiens" for the species, and set the confidence level to ≥0.9 to obtain the interaction relationship between proteins, and save the TSV and PNG files. Import them into the Cytoscape software, delete the isolated targets without intersection, analyze the network topology properties, and screen the core targets according to the degree value sorting (CC / BC / degree sorting) of the topological structure / use the MCC algorithm of the cytohubba plug-in to screen the core targets.

[0031] Import 195 intersection targets related to diseases treated by drugs into the STRING database to obtain the PPI relationship, and then import them 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 targets (delete the isolated targets without intersection). Use the Network Analyzer plug-in to analyze the network topology properties, calculate the degree value, set the depth of color and the size of the nodes to reflect the size of the degree value, and select the top 30 core nodes according to the degree value (CC / BC / degree) ranking to draw the network diagram, as shown in Figure 8 。The core targets are TP53, TNF, JUN, IL6, AKT1, STAT3, CYP3A4, BCL2, etc., which are used as the key targets for the hair-growing traditional Chinese medicine composition to treat hair loss, as shown in Figure 9 。

[0032] Experimental Example 5: GO Functional Enrichment and KEGG Pathway Enrichment Analysis The hair-growing traditional Chinese medicine composition and the related targets of hair loss were respectively subjected to gene ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis using the R clusterProfiler 4.6.2 package. For the GO enrichment analysis, with P < 0.05 as the screening condition, the top 20 biological terms with P values less than 0.05 in the results were selected to draw a bar chart. For the KEGG enrichment analysis, also with P < 0.05 as the significant enrichment screening condition, the top 30 signaling pathways with P < 0.05 were selected to draw a bubble chart for visual analysis.

[0033] Based on R clusterProfiler, GO enrichment analysis was performed on 195 intersection targets related to drug-treated diseases. The top 15 entries of biological process (BP), molecular function (MF), and cellular component (CC) with P < 0.05 were respectively selected to draw a bar chart, as Figure 10 , shown in Figure 11. The vertical axis represents each term of GO, and the horizontal axis represents the number of targets enriched to that term. BP mainly involves responses to external stimuli, responses to lipopolysaccharides, cellular responses to external stimuli, responses to nutritional levels, responses to extracellular stimuli, responses to bacterial molecules, responses to oxygen levels, gland development, responses to decreased oxygen levels, steroid metabolic processes, responses to peptides, responses to hypoxia, etc.; MF mainly includes heme binding steroid hydroxylase activity, tetrapyrrole binding, oxidoreductase activity acting on paired donors, monoxygenase activity, oxidoreductase activity acting on paired donors, nuclear receptor activity, ligand-activated transcription factor activity, aromatase activity, estrogen 16-α-hydroxylase activity, iron ion binding, etc. CC mainly involves transcriptional regulator complex membrane rafts, membrane microdomains, RNA polymerase II transcription regulator, vesicle lumen, protease inhibitor complex, outer leaflet of plasma membrane, cytoplasmic vesicle lumen, death-inducing signaling complex, caveolae, plasma membrane rafts, peptidase inhibitor complex, etc. KEGG pathway enrichment analysis was performed on the targets, and a total of 174 signaling pathways were enriched. In the present invention, the top 30 pathways with P < 0.05 in the downloaded results were selected to draw a bubble chart for visual analysis, as Figure 12, as shown in Figure 13. The enrichment results show that it mainly involves signal pathways such as lipid and atherosclerosis, diabetic AGE-RAGE signaling pathway, platinum resistance, chemical carcinogenesis-receptor activation, apoptosis, hepatitis B, prostate cancer, measles, Kaposi's sarcoma-associated herpesvirus infection, TNF signaling pathway, endocrine resistance, etc. These pathways communicate with each other and play an important role in the occurrence and development of diseases. In addition, the present invention also drew a Sankey diagram of the TOP pathways related to the TOP targets, as shown in Figure 14 .

[0034] Experimental Example 6: Construction and Analysis of Drug-Compound-Target-Pathway-Disease Network To discover the potential mechanism of drug action, the core TOP pathways and the key targets and active ingredients therein were selected to construct a "drug-compound-target-pathway-disease" network. In order to better display the mutual connection of the important active ingredients and targets of the hair-growing traditional Chinese medicine composition with the above important pathways and play a synergistic role.

[0035] The core TOP pathways and the key targets and active ingredients therein were selected to construct a "compound-target-pathway-disease" network. As Figure 15 shown, this network consists of 407 nodes and 3053 edges (168 components, 195 targets, 30 signal pathways, drugs and diseases). Different node shapes represent different active ingredients, key targets and regulatory pathways in the hair-growing traditional Chinese medicine composition, and the depth of color 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 hair-growing traditional Chinese medicine composition in preventing and treating hair loss with multiple components and multiple targets.

[0036] 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-growing 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 by removing solvent molecules and other operations with Pymol 2.6.0 software, and then further hydrogenated and charged with AutoDock Tools 1.5.7. The core target proteins and active compounds were saved as files in the "pdbqt" format, and the appropriate grid position and size were set. Finally, the docking of the components and targets was completed by Autodock Vina. The docking results were visualized with PyMOL software.

[0037] Combined with the ranking of the activity component values in the active ingredient-target network diagram and literature research, the present invention respectively performs molecular docking on the active ingredients MOL000006, MOL000098, MOL000173, MOL000422, MOL002039, MOL002714, MOL002773, MOL002776, MOL005030, MOL008691 in the hair-growing traditional Chinese medicine composition and the top 10 core targets AKT1, BCL2, CYP1A1, CYP3A4, ESR1, IL6, JUN, STAT3, TNF, TP53 in the PPI network. The docking results are shown in Figure 16 . The smaller the binding energy, the higher the binding activity, and the easier the compound binds to the target. A binding energy ≤ -5.0 kcal / mol indicates good binding, and a binding energy ≤ -7.5 kcal / mol indicates very good binding. The molecular docking uses PyMOL 2.6.0 software and Discovery Studio to visualize the docking results. The 3D and 2D diagrams of the docking of the Top6 compounds with the targets are shown in Figure 17 .

[0038] Experimental Example 8. Animal Experiment Experimental Animals and Grouping A total of 20 C57 mice were purchased from the Animal Medical Center of Yangzhou University. After the mice in each group were anesthetized by intraperitoneal injection of 7.5% chloral hydrate solution, a small electric hair clipper was used to carefully shave the hair on the back of the mice. The shaved area was approximately 3 cm × 4 cm. According to the instruction manual of the hair removal cream, the hair removal cream was applied to the skin area where the hair had been shaved. After waiting for 10 minutes, a cotton ball or gauze moistened with physiological saline was used to wipe off the hair removal cream and the remaining hair. At this time, it could be observed that the skin on the back of the mice showed a light pink color. It was divided into two experiments, with 10 mice in the hair-growing group and 10 mice in the blank group. The hair-growing group was smeared with the water decoction of the hair-growing traditional Chinese medicine composition in Example 3 at the hair removal site every day. Photos were taken on the 0th, 5th, and 10th days, and the mice were sacrificed and sampled after taking photos on the 10th day.

[0039] Result 1: Hair Growth Condition After hair removal treatment, starting from the 1st day after hair removal, corresponding experimental interventions were performed on the hairless areas on the backs of the mice in each group at 10:00 am every day, and the hair growth was observed. On the 0th, 5th, and 10th days after hair removal, the hairless areas on the backs of the mice were photographed. On the day of hair removal, the hairless areas on the backs of the mice in each group were pink or light gray, and there was no hair growth; on the 5th day after hair removal, it was observed that there was more hair growth in the mice in the hair growth promoting formula group, while there was only less hair growth in some areas in the blank group; on the 10th day, the hair of the mice in the hair growth promoting formula group had basically all grown out, and the new hair had a shiny feeling and was black and bright in color; the hair growth of the mice in the blank group was less, and there was even only light gray fluff, and there was an obvious boundary line at the edge of the old and new hair and skin. The hair growth at the hair removal boundary was slow. There were terminal hairs on the backs of the mice in the blank group and the normal saline group, but the growth condition was not as good as that in the hair growth promoting formula group ( Figure 18 )

[0040] Result 2: HE pathological staining On the 10th day of the experimental treatment, the skin of the mice was taken for pathological sections and stained with H&E to observe the hair growth of the mice. It was found that the number of hair follicles in the mice in the blank group was relatively small. By horizontal comparison, it was found that the hair growth in the hair growth promoting formula group was more than that in the blank group ( Figure 19 ) Figure 20 On the microscope (×20), the statistics and comparison of the number of hair follicles in each group (

[0041] Result 3: Immunohistochemical staining Immunohistochemical staining with Ki67 protein, a marker of cell proliferation index, was performed on paraffin sections of the skin tissues on the backs of the mice. The positive staining result was brown-yellow particles, and the darker the color, the more obvious it was, indicating that the more Ki67 protein was expressed, that is, the more vigorous the cell proliferation and division. The results showed that Figure 21 , the expression of Ki67 protein in the hair follicle cells of the skin tissues in the hair growth promoting formula group was significantly higher than that in the blank group, suggesting that the hair growth promoting formula promoted the proliferation activity of hair follicle cells after treatment.

[0042] Result 4: Immunofluorescence staining After the skin tissue specimens of the mice in each group were made into paraffin sections, immunofluorescence staining was performed. The marker was cell proliferation protein Ki67 (red), the cell nucleus was labeled with DAPI (blue), and apoptotic cells were stained with Tunel (green). The positive staining result of DAPI was blue, the positive staining result of Ki67 was red, and the positive staining result of Tunel was green. The darker the red staining of Ki67, the more obvious it was, indicating that the more Ki67 protein was expressed, that is, the more vigorous the cell proliferation and division; the darker the green staining of Tunel, the more cell apoptosis there was. 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 growth formula group was significantly higher than that of the blank group, and there were basically no apoptotic cells in the hair growth formula group, further confirming that the hair growth formula promoted the proliferation of hair follicle cells after treatment.

Claims

1. A hair growth Chinese medicine composition for preventing and treating hair loss, characterized in that: The invention is prepared from the following raw materials in proportion by weight: 5-20 parts of houttuynia cordata, 5-20 parts of scutellaria baicalensis, 80-150 parts of arborvitae leaves, 5-20 parts of sophora flavescens, 1-10 parts of cimicifuga heracleifolia, 5-20 parts of perilla frutescens, 5-20 parts of divaricate cortex, 3-20 parts of imperata rhizome, 5-20 parts of kochia scoparia, 3-20 parts of calamus tatarinowii, 1-10 parts of bupleurum, 1-10 parts of pulsatilla scabra and 1-10 parts of frankincense.

2. The hair growth Chinese medicinal composition for preventing and treating hair loss according to claim 1, characterized in that: The weight ratio of the raw materials is: 9 parts of houttuynia cordata, 9 parts of scutellaria baicalensis, 128 parts of arborvitae leaves, 12 parts of sophora flavescens, 3 parts of cimicifuga, 9 parts of perilla, 12 parts of dittany bark, 6 parts of imperata rhizome, 9 parts of kochia scoparia, 6 parts of calamus, 3 parts of bupleurum, 3 parts of pulsatilla and 3 parts of frankincense.

3. The hair growth 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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