A Chinese medicine extract composition for promoting hair follicle growth and its application

Abstract

The present invention discloses a Chinese medicine extract composition and application thereof for promoting hair follicle growth, wherein the Chinese medicine extract composition comprises by weight: 1-5 parts of safflower extract, 0.5-3 parts of ginseng extract, 1-5 parts of Polygonum multiflorum extract, 0.1-1 parts of Artemisia argyi extract, 1-2 parts of curcumin extract, 0.5-2 parts of Cynanchum chinense extract and 3-8 parts of peppermint extract. The Chinese medicine extract composition provided by the present invention can significantly improve the expression of hair papilla cell antioxidant genes (GCLM, HMOX1, FTH1, FTL, NQO1) and inhibit the expression of inflammatory factor IL-6, repair hair papilla cell oxidative damage, thereby significantly promoting hair follicle growth, and has a significant effect on alopecia caused by anxiety and postpartum depression.

Description

Technical Field

[0001] The invention relates to the technical field of drug formulations, and in particular to a Chinese medicine extract composition for promoting hair follicle growth and an application thereof. Background Art

[0002] Hair loss is one of the common dermatological diseases, with symptoms of sudden or gradual hair loss in one or more parts of the body. The number of patients is currently on the rise. Dermal papilla cells (DPC) play an important role in hair follicle morphogenesis and hair growth cycle. Therefore, exploring effective chemicals or natural extracts that can repair and stimulate the growth of DPC is of great significance for the treatment of hair loss.

[0003] Currently, only finasteride and minoxidil are approved by the Food and Drug Administration for the treatment of hair loss, and both have significant side effects. Given the limited efficacy and significant side effects of existing drugs, it is of great significance to find new natural anti-hair loss ingredients with fewer side effects and better effects. In recent years, some herbal topical formulations that prevent hair loss or promote hair growth have been launched around the world. At present, research on natural drugs that promote the growth of DPC mainly focuses on some plant extracts, such as safflower extract, hawthorn extract, geranium extract and Polygonum multiflorum extract. However, studies have found that a single plant extract has a poor effect on promoting hair growth, and the composition of different types of plant extracts has obvious differences in promoting hair growth.

[0004] Therefore, the prior art still needs to be improved and developed. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a Chinese herbal extract composition for promoting hair follicle growth and its application, aiming to solve the problem that the existing plant extract drug formula has a poor effect on promoting hair growth.

[0006] The technical solution of the present invention is as follows:

[0007] A Chinese herbal medicine extract composition for promoting hair follicle growth, wherein the Chinese herbal medicine extract composition comprises, by weight: 1-5 parts of safflower extract, 0.5-3 parts of ginseng extract, 1-5 parts of Polygonum multiflorum extract, 0.1-1 parts of Artemisia argyi extract, 1-2 parts of curcumin extract, 0.5-2 parts of Cypress extract and 3-8 parts of peppermint extract.

[0008] The Chinese herbal extract composition for promoting hair follicle growth comprises, by weight: 3.7 parts of safflower extract, 1 part of ginseng extract, 2 parts of Polygonum multiflorum extract, 0.4 parts of Artemisia argyi extract, 1.4 parts of curcumin extract, 1 part of Cypress extract and 6 parts of peppermint extract.

[0009] The Chinese herbal extract composition for promoting hair follicle growth, wherein the various Chinese herbal extract components in the Chinese herbal extract composition are prepared by the following steps:

[0010] Adding a Chinese medicine raw material into an ethanol solution at room temperature and performing sealed extraction for 70-80 hours to obtain an extract;

[0011] The extract is filtered, and the filtrate is concentrated using a vacuum rotary evaporator at 45-50° C. to obtain a viscous extract;

[0012] The viscous extract is frozen overnight and then subjected to vacuum freeze drying to obtain a traditional Chinese medicine extract.

[0013] An application of a Chinese herbal medicine extract composition, wherein the Chinese herbal medicine extract composition of the present invention is used to prepare a medicine for treating hair loss, and the medicine comprises a solvent and the Chinese herbal medicine extract composition dissolved in the solvent.

[0014] The application of the Chinese herbal extract composition, wherein the concentration of the drug is 250 μg·mL -1 .

[0015] Beneficial effects: The present invention studies the effects of safflower extract, ginseng extract, Polygonum multiflorum extract, wormwood extract, curcumin extract, cypress extract and peppermint extract on hair growth, and finds that when the above-mentioned Chinese herbal extracts are mixed in a specific ratio to form a composition, they have a significant effect on the proliferation and antioxidant capacity of DPC. The present invention also verifies the actual effect of the Chinese herbal extract composition provided by the present invention on promoting hair growth through mouse experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a graph showing the results of a DPC cell viability test treated with different drugs before and after oxidative stress in the present invention.

[0017] Figure 2 This is a graph showing the effects of the Chinese herbal extract composition of the present invention and minoxidil on the expression of DPC antioxidant genes.

[0018] Figure 3 This is a graph showing the effects of the Chinese herbal extract composition of the present invention and minoxidil on the expression of the inflammatory factor IL-6 in DPC. DETAILED DESCRIPTION

[0019] The present invention provides a Chinese herbal extract composition for promoting hair follicle growth and its application. In order to make the purpose, technical scheme and effect of the present invention clearer and more specific, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0020] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0021] The invention provides a traditional Chinese medicine extract composition for promoting hair follicle growth. The traditional Chinese medicine extract composition comprises, by weight, 1-5 parts of safflower extract, 0.5-3 parts of ginseng extract, 1-5 parts of polygonum multiflorum extract, 0.1-1 parts of wormwood extract, 1-2 parts of curcumin extract, 0.5-2 parts of juniper extract and 3-8 parts of peppermint extract.

[0022] In order to explore the effect of the Chinese herbal extract composition of the present invention on hair growth, the CCK-8 method was used to determine the effects of the Chinese herbal extract composition (safflower, ginseng, Polygonum multiflorum, wormwood, curcumin, cypress, and mint) on the proliferation and antioxidant capacity of dermal papilla cells (DPC), and RT-qPCR was used to detect the effect of the Chinese herbal extract composition of the present invention on the expression of key antioxidant and anti-inflammatory genes in DPC.

[0023] The results showed that the Chinese herbal extract composition (250 μg·mL-1) provided by the present invention can significantly promote DPC proliferation (p<0.001), significantly improve the antioxidant capacity of DPC (p<0.0001), significantly inhibit the expression of inflammatory gene IL-6 and promote antioxidant-related genes (GCLM, HMOX1, FTH1, FTL, NQO1) (p<0.05). Animal experiments and histological analysis showed that the Chinese herbal extract composition of the present invention shortened the resting phase of mouse hair follicles and had the effect of promoting hair growth.

[0024] In some embodiments, the Chinese medicine extract composition includes, by weight: 3.7 parts of safflower extract, 1 part of ginseng extract, 2 parts of Polygonum multiflorum extract, 0.4 parts of Artemisia argyi extract, 1.4 parts of curcumin extract, 1 part of Cyperus chinensis extract and 6 parts of peppermint extract. This embodiment provides a Chinese medicine extract composition with a specific compatibility ratio, and the Chinese medicine extract composition in this embodiment shows a better effect of promoting hair growth in subsequent experiments.

[0025] In some embodiments, the various Chinese medicine extract components in the Chinese medicine extract composition are prepared by the following steps: adding a Chinese medicine raw material to an ethanol solution at room temperature for sealed extraction for 70-80 hours to obtain an extract; filtering the extract, and then using a vacuum rotary evaporator to concentrate the filtrate at 45-50°C to obtain a viscous extract; freezing the viscous extract overnight, and then performing a vacuum freeze-drying process to obtain a Chinese medicine extract. In the extraction process of this embodiment, the mass ratio of the Chinese medicine raw material to the ethanol solution is preferably 1:10; the viscous extract is preferably frozen overnight in a refrigerator at minus 80°C.

[0026] In some embodiments, an application of a Chinese herbal extract composition is also provided, wherein the Chinese herbal extract composition of the present invention is used to prepare a drug for treating hair loss, and the drug includes a solvent and a Chinese herbal extract composition dissolved in the solvent. In this embodiment, the solvent is a safe and non-irritating water solvent or other organic solvent, for example, a mixed solvent consisting of 1,2-propylene glycol and ethanol in a volume ratio of 1:1.

[0027] In some specific embodiments, when the concentration of the drug is 250 μg·mL -1 When used, it has a better effect in promoting hair growth.

[0028] The present invention is further explained below by specific embodiments:

[0029] Example 1

[0030] Provided is a traditional Chinese medicine extract composition, which comprises, by weight, 3.7 parts of safflower extract, 1 part of ginseng extract, 2 parts of polygonum multiflorum extract, 0.4 parts of wormwood extract, 1.4 parts of curcumin extract, 1 part of juniper extract and 6 parts of peppermint extract.

[0031] Example 2

[0032] Provided is a traditional Chinese medicine extract composition, which comprises, by weight: 1 part of safflower extract, 0.5 parts of ginseng extract, 1 part of polygonum multiflorum extract, 0.1 parts of wormwood extract, 1 part of curcumin extract, 0.5 parts of juniper extract and 3 parts of peppermint extract.

[0033] Example 3

[0034] Provided is a traditional Chinese medicine extract composition, which comprises, by weight: 5 parts of safflower extract, 3 parts of ginseng extract, 5 parts of polygonum multiflorum extract, 1 part of wormwood extract, 2 parts of curcumin extract, 2 parts of cypress extract and 8 parts of peppermint extract.

[0035] Example 4

[0036] The CCK-8 method was used to detect the effect of the Chinese medicine extract composition in Example 1 on the antioxidant capacity of DPC:

[0037] The Chinese herbal extract composition in Example 1 was dissolved in 1,2-propylene glycol / ethanol (1:1, V / V) to prepare 25 μg mL -1 , 75 μg·mL -1 , 150μg·mL -1 , 250μg·mL -1 After adding to the cell culture medium for DPC treatment, the old medium was discarded, each well was rinsed twice with PBS, and then 100 μM H 2 O 2 Oxidative stress for 1.5 hours; after 1.5 hours, the old culture medium was aspirated, each well was rinsed twice with PBS, and then 100 μl DMEM and 10 μl CCK-8 were added to each well, incubated for 1 hour, and finally the absorbance was measured at 450 nm; the antioxidant capacity was expressed by comparing the viability of DPC cells treated with the same drug before and after oxidative stress; each sample was repeated three times. This embodiment also provides a control group and a minoxidil group, wherein the control group is not treated with DPC, and the viability of DPC cells treated with the same drug before and after oxidative stress is measured; the minoxidil group is treated with minoxidil on DPC, and the viability of DPC cells treated with the same drug before and after oxidative stress is measured.

[0038] Studies have shown that oxidative stress is one of the important factors leading to hair loss. Oxidative stress is caused by insufficient antioxidant defense or excessive production of free radicals. Long-term exposure of the skin to endogenous and environmental pro-oxidants leads to the production of reactive oxygen species (ROS). Excessive ROS can cause damage to cellular components such as proteins, nucleic acids, and cell membrane lipids. Therefore, DPCs were cultured with a combination of Chinese herbal extracts at different concentrations and then treated with H 2 O 2 Oxidative stress was conducted to explore the effect of the Chinese medicine composition on the antioxidant capacity of DPC. The results are as follows Figure 1 As indicated, 100 μM HO 2 O 2 The survival rate of DPC after oxidative stress for 1.5 h was 11.17% to 40.05% at a concentration of 250 μg mL -1 The mixture of Chinese herbal extracts had the best antioxidant effect, with a survival rate of 40.05% for DPC, which was 3.59 times that of the control group and 3.10 times that of the minoxidil group, while there was no significant difference between the control group and the minoxidil group. -1 The mixture of Chinese herbal extracts can significantly improve the survival rate of DPCs under oxidative stress.

[0039] Example 5

[0040] Real-Time Quantitative PCR (RT-RT-qPCR) was used to study the effect of the Chinese medicine extract composition in Example 1 on the expression of DPC-related genes:

[0041] The concentration was 250 μg mL -1 DPCs were treated with a combination of Chinese herbal extracts and 10 μM minoxidil, respectively, at 37°C and 5% CO 2 The cells were incubated in a humidified incubator for 24 h. 24 h after drug treatment, DPCs were collected and total RNA was extracted using TRIzol reagent (Solarbio, Cat#R1100). PrimeScript TM cDNA was synthesized using RT reagent kit and gDNA Eraser (Takara, Cat#RR047A). Premix Ex Taq TM Quantitative RT-RT-qPCR was performed using a QuantStudio II (Takara, Cat#RR820A) and ABI QuantStudio 6 Flex (ABI, USA). The cycling protocol was as follows: 1 cycle at 95°C for 30 s, followed by 40 cycles at 95°C for 5 s, 60°C for 30 s, and finally at 95°C for 15 s, 60°C for 1 min, and 95°C for 15 s. Relative gene expression was calculated as 2^ -△△Ct , where GADPH was used as the housekeeping gene for normalization.

[0042] The results of the cell experiment in Example 4 showed that the Chinese herbal extract composition can significantly improve the survival rate of DPC under oxidative stress. This example will use RT-qPCR to analyze the expression differences of some key antioxidant genes (GCLM, HMOX1, FTH1, FTL, NQO1) in DPC to explore the antioxidant mechanism of the Chinese herbal composition. Heme can further promote the production of reactive oxygen species (ROS) as a pro-oxidant, while heme oxygenase 1 (HMOX1, ENSG00000100292) can decompose heme and reduce the oxidative damage caused by heme. Proteins that chelate transition metals are indirect antioxidants because they inhibit H through Fenton chemistry. 2 O 2 Generate HO -, such as ferritin, including ferritin light chain (FTL, ID: ENSG00000087086) and ferritin heavy chain 1 (FTH1, ID: ENSG00000167996). The most abundant antioxidant in all cells is glutathione (GSH), an abundant tripeptide molecule that plays a key role in protecting cells from oxidative stress-induced cell damage and detoxifying xenobiotics and drug metabolism. Glutamate cysteine ​​ligase modifying subunit (GCLM, ENSG00000023909) is the key subunit of the first rate-limiting enzyme for the synthesis of GSH and plays an important role in the synthesis of GSH. NADPH quinone oxidoreductase 1 (NQO1, ENSG00000181019) can catalyze the two-electron reduction of quinone compounds in cells, prevent the generation of ROS, and thus protect cells from oxidative damage. The RT-qPCR results in this example are as follows. Figure 2 As shown in the figure, it can be seen that compared with the control group, the Chinese herbal extract composition can significantly increase the expression level of antioxidant genes (GCLM, HMOX1, FTH1, FTL, NQO1) in DPC, which are 6.03, 27.11, 4.35, 2.72 and 1.59 times that of the control group, and 2.63, 18.70, 3.92, 2.25 and 1.31 times that of the minoxidil group. The results show that the Chinese herbal extract composition can increase the expression of antioxidant genes in DPC, reduce the damage of oxidative stress to hair follicles, and the effect is more significant than minoxidil.

[0043] Example 6

[0044] Effect of Chinese herbal extract composition on the expression of inflammatory factor IL-6 in DPC:

[0045] IL-6 is a representative pro-inflammatory mediator. Studies have shown that IL-6 plays an important role in the inflammatory process caused by infection and injury. 2 O 2 Oxidative stress was applied to induce inflammation, and the expression of IL-6 in DPCs after oxidative stress was detected by RT-qPCR. Figure 3 As shown in the figure, the expression of IL-6 in the oxidative damage group was 1.60 times that of the control group, indicating that oxidative stress can cause DPC inflammation. In addition, the addition of the Chinese herbal extract composition can significantly inhibit the expression of IL-6 in DPC under oxidative stress, which is only 3.38% of the oxidative damage group, while the expression of IL-6 in the minoxidil group is 51.52% of the oxidative damage group. It can be seen that the Chinese herbal extract composition can reduce the expression of the inflammatory factor IL-6, reduce the damage caused by inflammation caused by oxidative stress, and thus protect hair follicles.

[0046] Example 7

[0047] Effects of Chinese herbal extract composition on hair growth in mice:

[0048] The Chinese medicine extract composition used in this animal example is dissolved in 1,2-propylene glycol / ethanol (1:1, V / V). After 15 mice were selected for adaptive feeding for one week, they were randomly divided into 3 groups, namely the control group, the positive control group (5% minoxidil), and the Chinese medicine group. Throughout the study, the mice were housed in a standard laboratory diet and free drinking water environment, and were subjected to a 12-hour / 12-hour light / dark cycle at a temperature of 22±2°C. At the beginning of the experiment, hair removal was first performed. The mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital, low-melting point paraffin was melted and evenly applied to the back of the mice, and the back hair was removed after the paraffin solidified. C57BL / 6 mice are animal models of the hair cycle. The hair of this mouse is black, and the melanocytes in the skin are distributed in the hair follicles. Only in the hair growth period can melanin be synthesized and secreted, and the melanin is transferred to keratinocytes, making the skin black; in the hair regression period, melanin synthesis decreases, and the skin gradually changes from black to gray; there is no melanin synthesis in the resting period, and the skin is pink. Histological verification shows that the changes in the skin color of the mice can reflect the different periods of the hair growth cycle. The control group did not receive any treatment after plucking. In the positive control group, 100 μl of 5% minoxidil was evenly applied to the plucking area of ​​the mice every day, while in the Chinese medicine group, 100 μl of the Chinese medicine composition solution was applied every day. The hair growth condition and skin color changes were observed, and photos were taken every 7 days for 14 days. After 14 days, the hair length and weight of the mice were measured. A handful of new hair was randomly drawn from each dosing area of ​​each rat, and the length of 5 hairs was measured with a vernier caliper. The average value was taken as the hair length value (mm) of the rats in the dosing area. Then carefully cut off all the hair in the dosing area, put it in a Petri dish, and weigh it as the weight (mg) of the hair in the dosing area with a one-tenth electronic balance.

[0049] In order to detect the actual effect of the Chinese medicine composition on hair growth, C57BL / 6 mice were used as an experimental model. The experiment set up a blank control group, a minoxidil group and a Chinese medicine composition group, with 5 mice in each group. The 5% minoxidil group was used as the positive control group.

[0050] Table 1 shows the evaluation results of hair growth in each group. The 5% minoxidil group and the Chinese medicine group grew a small amount of hair on the 7th day and the 4th day, respectively, which were 2 days and 5 days faster than the blank control group. In addition, the hair growth of the 5% minoxidil group and the Chinese medicine group was no different from that of normal mice on the 14th day and the 9th day, respectively.

[0051] Table 1. Evaluation of hair growth in each group

[0052]

[0053]

[0054] Note: "-" indicates no hair growth, "+" indicates very little hair growth, "++" indicates obvious hair growth, "+++" indicates the hair is in basic good condition, and "++++" indicates the hair is no different from normal.

[0055] Table 2 shows the statistics of hair growth cycle of mice in each group. The period when the mouse skin changes from red to gray is the resting phase of hair follicles.

[0056] Table 2 Duration of hair follicle resting phase in each group of mice

[0057] Group Time for skin to change from red to gray (days) Blank control group 8.33±0.91a 5% Minoxidil Group 6.83±0.53c Chinese Medicine Group 4.33±0.52b

[0058] As can be seen from Table 2, the resting period of the Chinese herbal extract combination group and the 5% minoxidil group was shortened by 4 days and 1.5 days compared with the blank control group, indicating that the Chinese herbal extract combination group can greatly shorten the resting period of hair follicles and promote hair growth.

[0059] Table 3 is a statistical table of the length and weight of new hair in mice 15 days after depilation

[0060] Group Hair length(mm) <![CDATA[Gross weight (g·cm -3 )]]> Blank control group 5.17±0.02a 0.049±0.0026a 5% Minoxidil Group 8.15±0.34b 0.058±0.0060b Chinese Herbal Combination 8.93±0.24b 0.073±0.0003b

[0061] As can be seen from Table 3, compared with the blank control group, the Chinese herbal extract composition can significantly increase the hair growth of mice by 1.72 times, and the hair weight is significantly increased by 1.49 times. It can be seen that the Chinese herbal extract composition can significantly promote hair growth.

[0062] In summary, the Chinese herbal extract composition provided by the present invention can repair oxidative damage to hair papilla cells by significantly increasing the expression of antioxidant genes (GCLM, HMOX1, FTH1, FTL, NQO1) in hair papilla cells and inhibiting the expression of inflammatory factor IL-6, thereby significantly promoting hair follicle growth and having a significant effect on alopecia caused by anxiety and postpartum depression.

[0063] It should be understood that the application of the present invention is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. Use of a Chinese herbal extract composition in the preparation of a medicament for treating hair loss, It is characterized in that The medicine consists of a solvent and a Chinese herbal medicine extract composition dissolved in the solvent, wherein the Chinese herbal medicine extract composition consists of 1-5 parts of safflower extract, 0.5-3 parts of ginseng extract, 1-5 parts of polygonum multiflorum extract, 0.1-1 parts of mugwort extract, 1-2 parts of curcumin extract, 0.5-2 parts of cypress extract and 3-8 parts of peppermint extract by weight; various Chinese herbal medicine extract components in the Chinese herbal medicine extract composition are prepared by the following steps: adding a Chinese herbal medicine raw material into an ethanol solution at room temperature for sealed extraction for 70-80 hours to obtain an extract; filtering the extract, and then using a vacuum rotary evaporator to concentrate the filtrate at 45-50°C to obtain a viscous extract; freezing the viscous extract overnight and then performing a vacuum freeze-drying treatment to obtain a Chinese herbal medicine extract.

2. The use of the Chinese medicine extract composition according to claim 1, It is characterized in that The concentration of the drug is 250 μg mL -1 .

3. The use of the Chinese medicine extract composition according to claim 1, It is characterized in that The Chinese medicine extract composition consists of 3.7 parts of safflower extract, 1 part of ginseng extract, 2 parts of Polygonum multiflorum extract, 0.4 parts of Artemisia argyi extract, 1.4 parts of curcumin extract, 1 part of Cypress extract and 6 parts of peppermint extract by weight.

4. A drug for treating hair loss, It is characterized in that The medicine consists of a solvent and a Chinese herbal medicine extract composition dissolved in the solvent, wherein the Chinese herbal medicine extract composition consists of 1-5 parts of safflower extract, 0.5-3 parts of ginseng extract, 1-5 parts of polygonum multiflorum extract, 0.1-1 parts of mugwort extract, 1-2 parts of curcumin extract, 0.5-2 parts of cypress extract and 3-8 parts of peppermint extract by weight; various Chinese herbal medicine extract components in the Chinese herbal medicine extract composition are prepared by the following steps: adding a Chinese herbal medicine raw material into an ethanol solution at room temperature for sealed extraction for 70-80 hours to obtain an extract; filtering the extract, and then using a vacuum rotary evaporator to concentrate the filtrate at 45-50°C to obtain a viscous extract; freezing the viscous extract overnight and then performing a vacuum freeze-drying treatment to obtain a Chinese herbal medicine extract.

5. The drug according to claim 4, It is characterized in that The Chinese medicine extract composition consists of 3.7 parts of safflower extract, 1 part of ginseng extract, 2 parts of Polygonum multiflorum extract, 0.4 parts of Artemisia argyi extract, 1.4 parts of curcumin extract, 1 part of Cypress extract and 6 parts of peppermint extract by weight.

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