CN113197911B - Application of ginsenoside composition in preparing medicine for preventing and treating alopecia by acting on hair follicle tissues - Google Patents
Application of ginsenoside composition in preparing medicine for preventing and treating alopecia by acting on hair follicle tissues Download PDFInfo
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Abstract
Firstly, the invention provides the application of the ginsenoside composition in preparing the medicine for preventing and treating alopecia by acting on hair follicle tissues. The ginsenoside composition consists of ginsenoside Rg1, ginsenoside Re and ginsenoside CK, wherein the ginsenoside Rg1 and the ginsenoside Re are main active ingredients, and the ginsenoside CK is an auxiliary ingredient. The invention also provides application of the ginsenoside composition in preparing a medicine for improving the activity of hair follicle tissues, and the medicine is directly applied or sprayed on scalp to promote the hair follicles in the telogen phase to enter the regeneration cycle and promote the hair growth. The invention further provides application of the ginsenoside composition in preparation of a medicine for improving the activity of the in vitro cultured hair follicle tissues, and the ginsenoside composition is prepared into a tissue culture solution to act on the in vitro cultured hair follicle tissues, so that the activity of the cultured hair follicle tissues can be improved, and the survival rate of transplanted hair follicles is obviously improved.
Description
Technical Field
The invention belongs to the field of medicines, and relates to a new application of a ginsenoside composition consisting of ginsenoside Rg1, ginsenoside Re and ginsenoside CK, in particular to an application of the ginsenoside composition in preparing a medicine for preventing and treating alopecia by acting on hair follicle tissues.
Background
The hair has the functions of keeping warm and protecting the scalp from being injured by the outside, and has important significance in beautifying the personal image. The average number of hair loss of a normal person is about 40-100 per day, the normal hair loss is the hair in the catagen and the telogen, and the normal number of hair can be maintained because the hair entering the catagen and the hair newly entering the growth phase are constantly in dynamic balance. Pathological hair loss refers to abnormal or excessive loss of hair for a number of reasons. Pathological abnormal hair loss, such as male baldness or female hairline retrogradation, is the failure of the hair follicle to enter the growth cycle due to long-term telogen under certain factors. In recent years, with the increasing social competition, the increasing survival pressure, the high fat diet, the reduction of sleep, the environmental impact and the use of hair-styling products such as hair dyeing and permanent wave, the patients with alopecia have the signs of youth, and the young and middle-aged people become the leading forces of alopecia. Alopecia which appears to be atraumatic and elegant often places an unreasonable mental and psychological burden on the patient. Patients with alopecia often have a very strong desire for treatment.
Therefore, the anti-alopecia and hair-growing medicines and cosmetics are widely concerned. At present, the common pilatory in the market of China mainly takes various hair-growing shampoos, health-care products, lotions decocted traditional Chinese medicines and the like as main materials, and achieves the effect of promoting hair growth by mainly improving microcirculation, but the common pilatory generally has poor effect. There are few drugs for treating pathological alopecia in clinic, and these drugs are effective only for a small proportion of patients, and may have certain toxic and side effects after long-term use. Although hair loss can be fundamentally solved by hair follicle tissue transplantation, the hair follicle tissue transplantation also has the problems of extremely high price and low survival rate of hair follicle tissues. Thus, there is still a great need for a safe and effective drug for preventing and treating hair loss and improving the survival rate of hair follicles during hair follicle transplantation surgery.
Ginseng is a rare medicinal material which is popular among China and foreign countries, and the use of ginseng for health preservation and disease treatment has been over 2000 years. However, how to clearly and specifically elucidate the mode and route of action of ginseng on skin, hair, etc., and to physically solve the problem of influence of ginseng on skin and hair, so as to maximally develop the use and commercial value of ginseng in the fields of medicine, health care products and cosmetics, has been a hot issue that is constantly being sought by both domestic and foreign scholars, well-known pharmaceutical companies and cosmetic enterprises. Research shows that ginseng has the effects of preventing and reducing alopecia and promoting hair growth and development, and has obvious effect, and the mechanism of preventing and treating alopecia is mainly as follows: the ginseng contains phytohormone related to estrogen, and the existence of the substances can improve the blood circulation of the head skin, regulate the immunity and achieve the effect of promoting the hair growth. Reducing apoptosis and degeneration of hair follicle keratinocytes in anagen phase. Research shows that ginseng and ginsenoside in the formula have the effects of promoting the proliferation of mesenchymal stem cells of hair follicles, promoting the growth of the hair follicles and prolonging the hair growth period of mice.
At present, there are also many researches on hair loss prevention and hair growth with ginsenoside components. For example, WO2020/262933A1 discloses that ginsenoside Rg4 or a mixture of ginsenosides Rg2, Rg4, Rg6 and Rh1 has the effect of promoting hair follicle growth. CN105250152A discloses an anti-hair loss and hair growth essence which comprises panaxatriol saponins Re, Rg1, Rg2 and Rh1 and also discloses that the anti-hair loss and hair growth essence can directly act on scalp, promote metabolism of scalp cells, expand capillary vessels, improve blood circulation and enable hair follicle tissues to obtain reasonable nutrition.
The inventor of the application also focuses on the research of ginsenoside on preventing and treating alopecia and improving the survival of hair follicle tissues for many years, and unexpectedly discovers that the combination of ginsenoside Rg1 and ginsenoside Re as main active ingredients and ginsenoside CK as an auxiliary ingredient in a specific ratio can obviously improve the curative effect of the ginsenoside Rg1 and ginsenoside Re as the main active ingredients and obviously reduce the using amount of the active ingredients in the research process, thereby developing a pharmaceutical composition which has simple components, safety and effectiveness and can prevent and treat alopecia by acting on the hair follicle tissues. In particular, no report on a pharmaceutical composition for preventing and treating alopecia and improving hair follicle tissues, which contains ginsenoside CK, has been found before.
Disclosure of Invention
The invention aims to solve the defects of the prior art, realize the development and utilization of natural products by adopting a modern medicine research method, combine a large amount of pharmacodynamic experiment screening, and provide the application of the ginsenoside composition prepared by combining the ginsenoside Rg1 and the ginsenoside Re as main active ingredients and the ginsenoside CK as an auxiliary ingredient in a specific proportion in preparing the medicine for preventing and treating alopecia and improving the survival of hair follicle tissues.
Specifically, the invention is realized by the following technical schemes:
in a first aspect, the present invention provides a use of a ginsenoside composition in the preparation of a medicament for preventing and treating hair loss by acting on hair follicle tissues, the ginsenoside composition consisting of ginsenoside Rg1, ginsenoside Re and ginsenoside CK, wherein the ginsenoside Rg 1: ginsenoside Re: the ratio of the ginsenoside CK is (2-8): (2-8): (0.5-2).
Alternatively, in the above uses, the ginsenoside Rg 1: ginsenoside Re: the ratio of the ginsenoside CK is 5: 4: 1.
alternatively, in the above use, the alopecia is one or more of androgenetic alopecia, neurogenic alopecia, endocrine alopecia, nutritional alopecia, physical alopecia, chemical alopecia, infectious alopecia, congenital alopecia, and seasonal alopecia.
In a second aspect, the present invention provides the use of a ginsenoside composition of the first aspect in the preparation of a medicament for increasing the activity of hair follicle tissue.
Alternatively, in the above use, the medicament promotes telogen phase hair follicles into the regenerative cycle, promoting hair growth.
Alternatively, in the use of the first and second aspects above, the medicament comprises a ginsenoside composition of the first and second aspects above and a pharmaceutically acceptable carrier.
Preferably, the medicament is a topical medicament. More preferably, the external medicine is in the form of a gel, ointment, cream, foam, shampoo, hair conditioner or spray type liquid preparation.
In a third aspect, the present invention provides the use of a ginsenoside composition of the first and second aspects in the preparation of a medicament for increasing the activity of hair follicle tissue cultured in vitro.
Alternatively, in the above use, the drug improves the microenvironment of hair follicle tissues, promotes the transition of the hair follicle tissues in the resting stage and the catagen stage to the hair follicle tissues in the anagen stage, and further improves the activity of the cultured hair follicle tissues and the survival rate of transplanted hair follicles significantly.
Alternatively, in the use of the third aspect described above, the medicament is contained in a hair follicle tissue culture solution which is generally used clinically.
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. For reasons of space, they will not be described in detail.
Compared with the prior art, the invention has the following beneficial effects:
the inventor unexpectedly finds that the combination of the main active ingredients of the ginsenoside Rg1 and the ginsenoside Re and the auxiliary ingredient of the ginsenoside CK in a specific proportion can obviously improve the curative effect of the main active ingredients of the ginsenoside Rg1 and the ginsenoside Re and obviously reduce the dosage of the active ingredients, thereby developing the pharmaceutical composition which has simple components, safety and effectiveness and can prevent and treat alopecia by acting on hair follicle tissues.
Pharmacodynamic experiments show that the ginsenoside CK has a synergistic effect on the ginsenoside Rg1 and the ginsenoside Re in the aspects of preventing and treating alopecia and stimulating hair regeneration. In addition, the ginsenoside composition has the effect of promoting the growth of mouse hair follicle hairs cultured in vitro, can promote the proliferation of hair follicle bulb cells, and has the effect remarkably superior to that of the ginsenoside composition (ginsenoside Rg1+ ginsenoside Re) without ginsenoside CK.
Detailed Description
For a better understanding of the present invention, the effects of the main raw and auxiliary materials in the composition of the present invention are further explained below.
As used herein, "auxiliary ingredient" generally refers to a substance that has little or no target pharmacological activity, but is capable of enhancing the target pharmacological activity of the primary active ingredient. The target pharmacological activity of the invention is mainly to prevent and treat alopecia by acting on hair follicle tissues, improve the activity of the hair follicle tissues, improve the activity of cultured hair follicle tissues and improve the survival rate of transplanted hair follicles.
The ginsenoside Rg1, the ginsenoside Re and the ginsenoside CK in the ginsenoside composition can be extracted and separated from plants such as ginseng containing the active ingredients by adopting conventional extraction and purification methods in the field of natural medicinal chemistry, and can also be purchased from commercial products.
The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and description, and is in no way intended to limit the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products which are not known to manufacturers and are available from normal sources.
Example 1: the ginsenoside composition has the effect of preventing and treating alopecia
In order to achieve the effect of preventing hair loss and promoting hair growth, 80 alopecia subjects with the age of 18-60 years, the average age of 30.5 years and half of men and women are selected. The above subjects suffering from alopecia were divided into 4 groups of 20 subjects each.
The following conventional hair care essences were administered to each group: (1) blank conventional hair care essence base: (2) the composition containing ginsenoside Rg1 and ginsenoside Re and the hair care essence of a conventional hair care essence matrix, wherein the ginsenoside Rg1 accounts for 5% of the total weight of the hair care essence in percentage by weight, and the ginsenoside Re accounts for 4% of the total weight of the hair care essence in percentage by weight; (3) the hair care essence contains ginsenoside CK and a conventional hair care essence matrix, wherein the ginsenoside CK accounts for 1% of the total weight of the hair care essence in percentage by weight; (4) the hair care essence comprises a composition containing ginsenoside Rg1, ginsenoside Re and ginsenoside CK and a conventional hair care essence matrix, wherein the ginsenoside Rg1 accounts for 5 wt% of the total weight of the hair care essence, the ginsenoside Re accounts for 4 wt% of the total weight of the hair care essence, and the ginsenoside CK accounts for 1 wt% of the total weight of the hair care essence.
The hair care essences of (1) to (4) are respectively directly smeared on the scalp of a subject, and are gently massaged for 5 minutes, once in the morning and at night and continuously used for 2 months. At the end of the study, the subjects were observed and counted for hair volume, hair density. The experimental results are shown in table 1 below.
Table 1: the ginsenoside composition has the effect of preventing and treating alopecia
As can be seen from the results in table 1, the composition comprising ginsenoside CK alone had little effect on preventing hair loss and stimulating hair regrowth, compared to the blank control group. The composition containing ginsenoside Rg1 and ginsenoside Re has certain effects of preventing and treating alopecia and stimulating hair regeneration, but the effect is far less than that of the composition containing ginsenoside Rg1, ginsenoside Re and ginsenoside CK. Therefore, in the aspect of preventing and treating alopecia, the ginsenoside CK has a synergistic effect on the ginsenoside Rg1 and the ginsenoside Re.
Example 2: the Effect of the ginsenoside composition on the hair growth and hair follicle bulb cell proliferation of the mouse hair follicle cultured in vitro
In vitro experiment of mouse tentacle hair follicle
1. Mouse tentacle hair follicle organ culture
Cervical dislocation of 4-6 week-old C57BL/6J mice was sacrificed, bilateral beards were cut off, the tentacles were sterilized with 75% alcohol 3 times, tentacle pads were cut off under aseptic conditions, washed in D-Hanks solution 3 times, then placed in Wi l iams medium, under a dissecting microscope, hair follicles in birth period were separated with micro scissors and tweezers, and 1/3 parts of the hair follicles were cut off. The hair follicle culture is carried out in 96-well plate, one hair follicle is placed in each well, 0.2mL of Wi L iams culture medium serum-free medium containing 10mg/L transferrin, 100U/mL penicillin, 100 μ g/mL streptomycin and the like is added into each well, and the mixture is placed at 37 ℃ and 5% CO2Cultured in a cell culture box.
2. Experiment grouping
The experiment is divided into a blank group, an Rg1+ Re group, a CK group and an Rg1+ Re + CK group, each group has 12-14 hair follicles, and the blank group is not added with medicines. In the group Rg1+ Re, the final concentration of ginsenoside Rg1 is 1 × 10-5g/mL, ginsenoside Re final concentration of 8 × 10-6g/mL. The final concentration of ginsenoside CK in CK group is 2 × 10-6g/mL. In the group of Rg1+ Re + CK, the final concentration of ginsenoside Rg1 is 1 × 10-5g/mL, ginsenoside Re final concentration of 8 × 10-6g/mL, and the final concentration of the ginsenoside CK is 2 multiplied by 10-6g/mL。
3. Observation of Hair growth
Culturing on days 1 to 7, measuring the length from the bottom of the hair follicle to the top of the hair each day by an ocular micrometer under an inverted optical microscope, subtracting the measurement result of day 1 from the measurement result of day 7 to obtain the hair growth length, and comparing the hair growth lengths of the groups.
Mouse hair follicle bulb cell in vitro experiment
1. Mouse hair follicle bulb cell in vitro culture
Collecting the whole skin of 4d newborn C57BL/6J mice under aseptic condition, digesting with two-step enzyme (containing 0.5% separating enzyme and 0.2% collagenase type I) to obtain high-purity follicular bulbar cells, and inoculating to the culture solution of Wi l iams of blank group, Rg1+ Re group, CK group, Rg1+ Re + CK group with 6 wells (0.2 mL per well) and cell concentration of 5 × 104one/mL. Adding 5% CO at 37 deg.C2Cultured in a cell culture box.
2MTT colorimetric method for detecting influence of ginsenoside composition on growth activity of mouse hair follicle bulb cells
MTT colorimetric assay was performed 24h and 96h after the incubation, 20. mu.L of MTT solution (5mg/mL) was added to each well, incubation was continued at 37 ℃ for 4h, and the culture medium in the wells was discarded. 0.2mL of DMSO was added to each well, and the crystals were dissolved thoroughly by shaking 10 min. The wavelength is 570nm, the absorbance value of each hole is measured on a microplate reader, and the absorbance values of all groups are compared.
Statistical method
Results were analyzed using SPSS 20.0 statistical software and data are presented as mean ± SD. The comparison among multiple groups adopts one-factor analysis of variance, the comparison between two groups adopts t test, and p is less than 0.05 to indicate that the difference has statistical significance.
Results of the experiment
1. Hair growth length
The average growth length of hair follicles of each group on day 7 of culture is shown in table 2. The average hair follicle growth length of the Rg1+ Re + CK group was significantly greater than that of the blank group (p <0.05), although the Rg1+ Re group was also able to promote hair follicle growth in mice, but the difference from the blank group was not significant. There was no statistical difference in hair follicle hair growth length between CK and blank groups. It is suggested that Rg1 and Re in combination with CK significantly increased the ability of the two to promote the growth of the mouse whisker hair follicle.
2. Cell growth viability
MTT colorimetry also see table 2 for viability of follicular bulb cells. After 24h of culture, the average absorbance values among the blank group, the Rg1+ Re group, the CK group and the Rg1+ Re + CK group have no statistical significant difference, and the cell proliferation degrees among the groups are similar.
At 96h post-incubation, the average absorbance values between the CK and blank groups were similar, suggesting that the cell growth viability was similar in both groups. The average absorbance values of the Rg1+ Re group were higher than those of the blank group, but the difference between the two groups was not significant, and the average absorbance values of the Rg1+ Re + CK group were significantly higher than those of the blank group (p <0.05), suggesting that the combination of Rg1 and Re with CK can significantly increase the viability of the former two groups for promoting the growth of the bulbar follicular cells of mice.
Table 2: the ginsenoside composition of the invention has the effects on the growth of the hair follicle of the tentacle of a mouse and the growth activity of the hair follicle bulb cells of the mouse
Note that: p <0.05, compared to blank group.
Conclusion of the experiment
The ginsenoside composition (Rg1+ Re + CK group) has the function of promoting the growth of mouse hair follicle hairs cultured in vitro, can promote the proliferation of hair follicle bulb cells, and has the effect remarkably superior to the effect of the combination of the ginsenoside Rg1 and the ginsenoside Re after the ginsenoside CK is added.
In addition, ginsenoside CK had little effect on hair growth of mouse hair follicles and growth activity of mouse hair follicle bulb cells, compared to the blank group.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (9)
1. The application of the ginsenoside composition in preparing the medicine for preventing and treating alopecia by acting on hair follicle tissues is characterized in that: the ginsenoside composition consists of ginsenoside Rg1, ginsenoside Re and ginsenoside CK, wherein the ginsenoside Rg 1: ginsenoside Re: the ratio of the ginsenoside CK is 5: 4: 1.
2. the use according to claim 1, wherein the hair loss is one or more of androgenetic hair loss, neurogenic hair loss, endocrine hair loss, nutritional hair loss, physical hair loss, chemical hair loss, infectious hair loss, congenital hair loss, and seasonal hair loss.
3. Use of a ginsenoside composition of claim 1 in the preparation of a medicament for increasing hair follicle tissue activity.
4. The use of claim 3, wherein the medicament promotes the entry of telogen-phase hair follicles into the regenerative cycle, promoting hair growth.
5. The use according to claim 1, wherein the medicament comprises a ginsenoside composition of claim 1 and a pharmaceutically acceptable carrier.
6. The use according to claim 5, wherein the medicament is a topical medicament in the form of a gel, ointment, cream, foam, shampoo, conditioner or spray-on liquid.
7. Use of a ginsenoside composition of claim 1 in the preparation of a medicament for increasing the activity of in vitro cultured hair follicle tissue.
8. The use of claim 7, wherein the medicament improves the microenvironment of the hair follicle tissue, promotes the transition of the resting and catagen states to anagen hair follicle tissue, and thereby increases the viability of the cultured hair follicle tissue and significantly increases the survival rate of the transplanted hair follicle.
9. Use according to claim 7 or claim 8, wherein the medicament is contained in a clinically common hair follicle tissue culture fluid.
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| CN114948932B (en) * | 2022-06-13 | 2023-05-30 | 浩溪(海南)生物科技有限公司 | Use of a composition comprising mulberenone for the preparation of a product for the prevention and treatment of hair loss by acting on the hair follicle tissue |
| CN117414309B (en) * | 2023-02-23 | 2024-06-25 | 茵素科技(广州)有限公司 | Ginsenoside composition, daily chemical and application |
| CN116459176B (en) * | 2023-02-23 | 2024-01-30 | 茵素科技(广州)有限公司 | Ginsenoside-containing composition, daily chemical and application |
| CN116808054A (en) * | 2023-04-25 | 2023-09-29 | 三蚁科技(广州)有限公司 | A saponin composition for preventing alopecia, and its preparation method and application |
| CN116763716B (en) * | 2023-08-21 | 2023-10-24 | 成都中医药大学 | Bletilla polysaccharide microneedle carrying ginsenoside Rg3 and preparation method and application thereof |
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| CN110090236B (en) * | 2018-01-29 | 2022-09-16 | 吉林海颐康达生物科技发展有限公司 | Pharmaceutical composition and use thereof |
| KR102250224B1 (en) * | 2019-06-25 | 2021-05-10 | 아레즈 주식회사 | A pharmaceutical composition comprising ginsenoside Rg4 or ginsenoside Rg2, Rg4, Rg6 and Rh1 mixture(Rgx 365) for preventing hair loss or stimulating hair growth |
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