Use of a polysaccharide extract for the preparation of a medicament for hair growth

Abstract

The application discloses a use of a polysaccharide extract in preparation of a hair-growing medicine, and belongs to the field of medicines.The polysaccharide extract is angelica polysaccharide extract or fingered citron polysaccharide extract or a combination of both.The application is derived from Chinese herbal medicines, has the advantages of small side effects and high safety, and can be used to prepare medicines.Experiments prove that the polysaccharide extract has good hair-growing effects, can increase capillary neogenesis, increase the number of hair follicles, stimulate the activation of dormant hair follicles and promote the proliferation of hair papilla cells, so that the purpose of hair growth is achieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a use of a polysaccharide extract in preparation of a hair growth medicine. BACKGROUND

[0002] With the development of economic society, the social pressure faced by people is increasing, and the alopecia phenomenon is gradually becoming younger, and young people pay more and more attention to hair health. There are many reasons for alopecia, the most common of which are androgenetic alopecia and telogen effluvium. The growth of hair is generally divided into three periods: anagen, catagen and telogen. Telogen effluvium is one of the most common alopecia diseases, and often has an inciting cause. There are many reasons for telogen effluvium, such as postpartum, high stress, staying up late, malnutrition, local scalp inflammation and the like.

[0003] In order to combat alopecia, the common measures at present are to prevent the damage of hair follicle cell activity and prevent the shortening of the growth cycle of hair follicles: such as blocking the stimulation of androgens to hair follicles, increasing local blood flow to deliver more nutrients to hair follicle cells, inhibiting the stimulation of microinflammation to hair follicle cells and the like. Although there are many drugs for treating alopecia on the market, most of them have varying degrees of side effects, for example, the most well-known minoxidil and finasteride, and excessive use will bring about rapid heart rate, local inflammation, androgenic hypofunction and the like. SUMMARY

[0004] In order to overcome the above defects, the present application provides a use of a polysaccharide extract in preparation of a hair growth medicine, the polysaccharide extract is derived from Chinese herbal medicine angelica or finger citron, has the advantages of small side effects and high safety, and can be used as a medicine, and experiments prove that the polysaccharide extract has good hair growth effect.

[0005] The above object is achieved by the following technical solutions.

[0006] A use of a polysaccharide extract in preparation of a hair growth medicine, the polysaccharide extract is angelica polysaccharide extract or finger citron polysaccharide extract or a combination of the two.

[0007] Preferably, the polysaccharide extract is a combination of angelica polysaccharide extract and finger citron polysaccharide extract, and the mass ratio of the angelica polysaccharide extract to the finger citron polysaccharide extract is 1-5:10-15.

[0008] Further preferably, the mass ratio of the angelica polysaccharide extract to the finger citron polysaccharide extract is 3:14.

[0009] The angelica polysaccharide extract and the finger citron polysaccharide extract are extracted by a water extraction and alcohol precipitation method known in the art.

[0010] The test results show that the polysaccharide extract of angelica or bergamot can promote hair growth at the hair loss site of the mouse, accelerate the transformation of the hair follicle from the regression period to the growth period, increase the number of hair follicles and melanocytes, and increase the expression of vascular endothelial growth factor (VEGF) in the skin. And the polysaccharide extract of angelica combined with the polysaccharide extract of bergamot can produce better efficacy.

[0011] The beneficial effects of the present application are:

[0012] The present application provides a use of an extract in the preparation of a hair growth medicine. Compared with the existing hair growth medicine, the extract is derived from natural plants, has the advantages of small side effects and high safety. The present application further improves the efficacy by screening the plant species, extract category and formula, and the prepared medicine can increase capillary neovascularization, increase the number of hair follicles, stimulate the activation of dormant hair follicles, promote hair papilla cell proliferation, etc., and has high hair growth activity. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 : Skin and hair condition of animals in each group.

[0014] Figure 2 : The structure of the hair follicle of the blank group mouse observed by skin section.

[0015] Figure 3 : The structure of the hair follicle of the model group mouse observed by skin section.

[0016] Figure 4 : The structure of the hair follicle of the angelica group mouse observed by skin section.

[0017] Figure 5 : The structure of the hair follicle of the bergamot group mouse observed by skin section.

[0018] Figure 6 : The structure of the hair follicle of the polysaccharide composition group mouse observed by skin section.

[0019] Figure 7 : The structure of the hair follicle of the medicinal composition group mouse observed by skin section.

[0020] Figure 8 : The content of vascular endothelial growth factor in the skin tissue is detected by enzyme-linked immunosorbent assay. DETAILED DESCRIPTION

[0021] The present application will be described in detail below in combination with specific examples. The polysaccharide extract used in the present application can be obtained by commercial means. The polysaccharide extract used in the present embodiment is self-made by the water extraction and alcohol precipitation method well known in the art, and the specific method is as follows:

[0022] Preparation of Angelica polysaccharide extract: Fresh Angelica was dried at 40°C, crushed and sieved through a 60-mesh sieve. Distilled water was then added to the Angelica powder for ultrasonic extraction, with a solid-liquid ratio of 1:20 (g / ml), ultrasonic power of 500 W, temperature of 40°C and time of 1 h. After ultrasonic extraction, centrifugation was performed, and the supernatant was collected. The remaining residue was extracted twice more. The extraction solutions were combined, concentrated by rotary evaporation at 55°C, and then 4 times the volume of absolute ethanol was added to the concentrated solution. After stirring, the mixture was stored overnight, centrifuged, and the precipitate was collected, dried at 50°C until the weight was constant, and then freeze-dried to obtain the Angelica polysaccharide extract.

[0023] Preparation of Bergamot polysaccharide extract: Fresh Bergamot was dried at 50°C, crushed and sieved through a 60-mesh sieve. 95% ethanol was then added to a round-bottom distillation flask, and the mixture was refluxed at 70°C for 2 h. This process was repeated twice to remove fat. The dried medicinal material was then added to distilled water, and the mixture was extracted at 90°C for 3 times, each time for 2 h. After centrifugation, the supernatant was collected, concentrated by rotary evaporation at 50°C, and then 4 times the volume of absolute ethanol was added to the concentrated solution. After stirring, the mixture was alcohol precipitated for 12 h. After centrifugation, the ethanol supernatant was removed, and the polysaccharide precipitate was retained. The mixture was vacuum dried at 50°C to obtain the Bergamot polysaccharide extract.

[0024] The polysaccharide content in the two extracts was detected. The total polysaccharide content in the Angelica extract was 6.35%, and the total polysaccharide content in the Bergamot extract was 14.26%. The detection method was as follows: The standard curve was prepared using the phenol-sulfuric acid method. The mass concentration of glucose solution was used as the horizontal coordinate, and the absorbance was used as the vertical coordinate to draw the standard curve. The regression equation was Y=0.119x-0.01, R=0.996. The mass concentration of polysaccharide in the sample solution was calculated from the standard curve, and the polysaccharide content of the sample was calculated according to the following formula. Polysaccharide content = (p x V) x 100% / m. In the formula, p is the mass concentration of polysaccharide in the sample solution, g / ml; V is the volume of the sample solution, ml; and m is the mass of the sample, g. 2 =0.996. The mass concentration of polysaccharide in the sample solution was calculated from the standard curve, and the polysaccharide content of the sample was calculated according to the following formula. Polysaccharide content = (p x V) x 100% / m. In the formula, p is the mass concentration of polysaccharide in the sample solution, g / ml; V is the volume of the sample solution, ml; and m is the mass of the sample, g.

[0025] Experimental Example 1: Effect of polysaccharide extract on hair growth in model mice

[0026] 1. Test materials

[0027] Test animals: C57 / BL6 mice, male, 5 weeks old, a total of 36, purchased from the Animal Experiment Center of Huazhong Agricultural University, fed with free access to food and water, light according to 12h / 12h alternation, quarantine for 7 days before entering the experiment.

[0028] Test drugs: Angelica polysaccharide extract and Bergamot polysaccharide extract were prepared according to the above methods;

[0029] Polysaccharide composition: Angelica polysaccharide extract and Bergamot polysaccharide extract were mixed in a weight ratio of 3:14;

[0030] Medicinal material composition: Angelica sinensis and Citrus bergamia were crushed into fine powder, and then mixed evenly according to a weight ratio of 3:14.

[0031] 2. Test method

[0032] (1) Modeling: The mice were anesthetized with 10% chloral hydrate, and the anesthetic dose was 0.04 ml / 10 g. After about 5 minutes, the mice were deeply anesthetized, and the skin on the back of the mice was removed with an electric shaver. Then, the mice were smeared with depilatory cream on the back, about 2 cm*3 cm in size, and the depilatory cream was wiped off after about 2 minutes. The mice were confirmed to be successfully modeled when the skin color on the back was pink and there was no scar.

[0033] (2) Grouping and administration: The mice after modeling were randomly divided into 5 groups, the model group, the Angelica sinensis group, the Citrus bergamia group, the polysaccharide combination group, and the medicinal material composition group, with 6 mice in each group, and a total of 30 mice. The Angelica sinensis group was given Angelica sinensis polysaccharide extract by gavage at 5 mg / kg, the Citrus bergamia group was given Citrus bergamia polysaccharide extract by gavage at 5 mg / kg, the polysaccharide combination group was given polysaccharide combination by gavage at 5 mg / kg, and the medicinal material composition group was given medicinal material composition by gavage at 5 mg / kg. Gavage was performed once a day for 20 consecutive days, and the mice were fed with ordinary feed and allowed to drink water freely.

[0034] Another 6 normal mice were taken, and they were always fed with ordinary feed and allowed to drink water freely during the experiment, serving as the blank control group.

[0035] (3) Test method

[0036] The skin and hair condition of the mice were photographed regularly during gavage. After the last administration, the skin of the experimental area on the back of the mice was separated, 2*2 cm, and fixed with 4% paraformaldehyde for 24 hours. After dehydration and paraffin embedding, 4 μm paraffin sections were prepared, and after conventional dewaxing to water, hematoxylin-eosin staining was performed. The hair follicle structure of the full-thickness skin was observed under a light microscope. A part of the skin was weighed after removing blood and fascia, and PBS solution pre-cooled at 4°C was added at a ratio of 1:5. The skin was cut into small pieces and added with steel balls, and then put into a homogenizer to prepare skin tissue homogenate. The tissue homogenate was centrifuged at 5000 g for 10 minutes, and the supernatant was detected for the content of vascular endothelial growth factor by enzyme-linked immunosorbent assay (ELISA) kit.

[0037] 3. Test results

[0038] (1) Skin and hair condition of animals in each group

[0039] Test results: From Figure 1 it can be seen that the hair growth of the model group was slow, and only a small amount of hair grew within 20 days. The hair growth rate of the Angelica sinensis group, the Citrus bergamia group, the polysaccharide combination group, and the medicinal material composition group increased compared with the model group, especially the polysaccharide combination group, which grew very dense hair on the 20th day, and its effect was better than that of the other test groups. See Figure 1 .

[0040] (2) The hair follicle structure of each group of animals

[0041] Test results: from Figure 2 It can be observed that the skin section of the blank group mouse has no mature hair follicle tissue, the hair follicle vacuole phenomenon is serious, the hair follicle is in the resting period, and there is no new hair.

[0042] From Figure 3 It can be observed that the hair follicle vacuole phenomenon of the model group mouse is improved, and the number of hair follicles increases, which indicates that shaving the mouse hair helps to stimulate the hair follicle to change from the regression period to the growth period.

[0043] From Figures 4-5 It can be observed that the number of hair follicles in the angelica group and the bergamot group increases significantly, and the hair follicle cells are elongated, indicating that the hair follicle is in the growth period. Not only that, there are a large number of melanocytes around the mammary gland tissue, which is an accessory of hair and hair follicle.

[0044] From Figure 6 It can be observed that the hair follicle of the polysaccharide composition group mouse has penetrated into the dermis layer, and the hair follicle is in the vigorous period.

[0045] From Figure 7 It can be observed that the number of hair follicles in the medicinal material composition group increases, but the hair growth effect is not as good as the polysaccharide composition.

[0046] In summary, the polysaccharide extract and its combination provided by the present application have a hair growth effect.

[0047] (3) Expression of vascular endothelial growth factor (VEGF) in the skin

[0048] Test results: from Figure 8 It can be seen from the table that the bergamot polysaccharide group has no significant difference compared with the model control group (P>0.05), the polysaccharide composition group and the angelica polysaccharide group have extremely significant difference compared with the model control group (P<0.01), and the medicinal material composition group has significant difference compared with the model control group (P<0.05). And the polysaccharide composition group and the angelica polysaccharide group also have significant difference (P<0.05).

Claims

1. A use of a polysaccharide extract in the preparation of a hair growth and development drug, wherein the polysaccharide extract is a combination of an angelica polysaccharide extract and a bergamot polysaccharide extract, wherein the mass ratio of the angelica polysaccharide extract to the bergamot polysaccharide extract is 3:

14. Preparation of Angelica polysaccharide extract: fresh Angelica sinensis was dried at 40°C, crushed and passed through a 60-mesh sieve, and then distilled water was added to the Angelica sinensis powder for ultrasonic extraction at a solid-liquid ratio of 1:20, g / ml, ultrasonic power of 500W, temperature of 40°C, and time of 1 hour. After the end of the ultrasonication, the mixture was centrifuged and the supernatant was collected. The remaining medicinal residue was extracted twice again, and the extracts were combined and concentrated by rotary evaporation at 55°C. Then, 4 times the volume of anhydrous ethanol was added to the concentrate, stirred and stored overnight, centrifuged, and the precipitate was collected and dried at 50°C to constant weight. The precipitate was then freeze-dried to obtain the Angelica polysaccharide extract; Preparation of bergamot polysaccharide extract: fresh bergamot is dried at 50℃, crushed and passed through a 60-mesh sieve, then added with 95% ethanol in a round-bottom distillation flask, refluxed at 70℃ for 2h, repeated twice for defatting, then the medicinal material is dried, added with distilled water, extracted 3 times at 90℃ for 2h each time, centrifuged, the supernatant is collected, concentrated by rotary evaporation at 50℃, and then 4 times the volume of anhydrous ethanol is added to the concentrate, stirred and precipitated for 12h, centrifuged, the ethanol supernatant is removed, the polysaccharide precipitate is retained, and vacuum dried at 50℃ to obtain bergamot polysaccharide extract.

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