Ganoderma lucidum microsphere and algae co-fermentation technology with hair follicle stem cell repairing effect and hair growth promoting effect and application of ganoderma lucidum microsphere and algae co-fermentation technology

Abstract

The invention relates to an innovative method combining a modern biotechnology and a traditional Chinese medicine theory, and aims to improve the content of effective components in lucid ganoderma through optimization of a specific seaweed co-fermentation technology and a fermentation process, and promote repair and regeneration of hair follicle stem cells by using the active components so as to promote hair growth. According to the technology, ganoderma lucidum mycelia are inoculated into a specific culture medium to form a microsphere structure, and the structure is beneficial to exchange of nutrient substances and discharge of metabolic waste, so that the fermentation efficiency and the product quality are improved. Micrococcus fermentation can simulate the symbiotic relationship of microorganisms in the natural environment and promote ganoderma lucidum to generate more beneficial secondary metabolites. The invention further provides application of the product prepared on the basis of the technology in development of novel anti-hair loss and hair growth products, anti-aging skin care products and health supplements. Experimental results show that the technology can effectively promote mouse hair growth and significantly improve the expression level of growth factors in hair follicle stem cells, and shows wide application potential in biological medicine, beauty and skin care and large health industries.

Description

Technical Field

[0001] The present invention relates to the field of medical care. The Ganoderma lucidum Microchlorella fermentation technology for repairing hair follicle stem cells is an innovative method that combines modern biotechnology and traditional Chinese medicine theory. It aims to increase the content of active ingredients in Ganoderma lucidum through specific seaweed co-fermentation technology and optimization of fermentation process, and utilize these active ingredients to promote the repair and regeneration of hair follicle stem cells. Background Art

[0002] Ganoderma lucidum is a Polyporaceae fungus whose fruiting bodies are widely used in traditional Chinese medicine. It is known as "immortal grass" and has a wide range of pharmacological effects, such as enhancing immunity, anti-oxidation, anti-inflammatory, and anti-tumor. Its main components include polysaccharides, triterpenoids, peptides, etc., which have significant effects on cell proliferation, differentiation, and migration. Hair follicle stem cells play a vital role in the physiological functions of skin and hair. They are not only responsible for the growth and regeneration of hair, but also have an important impact on the health and repair of the skin. Hair follicle stem cells are located in the bulge area of ​​the hair follicle, and they are an indispensable cell group in the hair growth cycle. These stem cells can differentiate into different types of cells, such as keratinocytes, pigment cells, etc., thereby driving the hair into the growth phase, regression phase, and resting phase. Through continuous division and differentiation, hair follicle stem cells ensure the continuous growth and renewal of hair. With age, the number of stem cells in the human body will gradually decrease, and the activity will also decrease. This will lead to problems such as skin aging and increased wrinkles. However, hair follicle stem cells have anti-aging effects. They can promote the proliferation and differentiation of skin cells by secreting a variety of growth factors and cytokines, thereby slowing down the skin aging process. Since hair follicle stem cells are closely related to hair growth, they have potential application value in the treatment of hair loss and baldness. By activating or transplanting hair follicle stem cells, hair growth and regeneration can be promoted, thereby improving hair loss problems.

[0003] The co-cultivation and fermentation technology of Ganoderma lucidum microspheres and algae involves inoculating Ganoderma lucidum mycelium into a specific culture medium to form a microsphere structure, which is conducive to the exchange of nutrients and the discharge of metabolic waste, thereby improving fermentation efficiency and product quality. Microspherical algae fermentation can simulate the symbiotic relationship of microorganisms in the natural environment and promote Ganoderma lucidum to produce more beneficial secondary metabolites. After fermentation, the active ingredients in Ganoderma lucidum can be enhanced, thus bringing more significant medicinal effects. Ganoderma lucidum fermentation products can promote and adjust immune function and enhance the body's disease resistance; they also have significant anti-free radical effects, which help to remove harmful substances in the body; and they can promote the synthesis of nucleic acids and proteins, helping to maintain the body's metabolic balance; in addition, Ganoderma lucidum fermentation products also have significant anti-allergic effects, anti-neurasthenia and insomnia, and treatment of hypertension and diabetes.

[0004] It has been verified that products prepared based on the fermentation technology of Ganoderma lucidum microalgae can be used to develop new anti-hair loss products, which can promote hair growth and improve hair loss by activating hair follicle stem cells. In addition, Ganoderma lucidum extract can be used in anti-aging skin care products due to its good antioxidant and anti-inflammatory properties to help repair damaged skin and slow down signs of aging. In the future, microalgae fermentation products rich in Ganoderma lucidum polysaccharides and triterpenoids can also be used as health supplements to enhance immunity, regulate blood sugar and blood lipids, etc. In summary, Ganoderma lucidum microalgae fermentation technology is an innovative field of interdisciplinary integration. It not only provides new scientific basis and solutions for solving the problem of hair loss, but also opens up new ways for the modern use of traditional Chinese medicinal materials. In the future, with the deepening of research and the continuous optimization of technology, this technology is expected to play an important role in biomedicine, beauty and skin care, and the health industry. Summary of the invention

[0005] The problem to be solved by the present invention is to provide a co-fermentation technology of Ganoderma lucidum microspheres and algae which has the function of repairing hair follicle stem cells and promoting hair growth, so that it has the function of repairing hair follicle stem cells and promoting the regeneration of hair on the back of mice.

[0006] In order to solve the above technical problems, the present invention adopts the following technical solutions: On the one hand, a fermentation method of a fermentation liquid of Ganoderma lucidum microspheres and algae co-fermentation having the function of repairing hair follicle stem cells and promoting hair growth comprises the following steps: (1) Add potato dextrose medium (PDB) powder to a 1L conical flask and dilute to 1L with deionized water. Sterilize with high pressure steam at 0.103MPa and 121℃ for 20-30 minutes and cool to room temperature for subsequent use.

[0007] (2) Add the PDB culture medium powder to a 1L conical flask, add 10g agarose gel, and use deionized water as the solvent to make the volume 1L. Sterilize with high pressure steam at 0.103MPa and 121℃ for 20-30 minutes. After the culture medium cools to 50-55℃, pour it into a round flat plate culture dish in the clean bench, 20-25ml per dish, and cool to room temperature until solidified.

[0008] (3) Use sterile tweezers to pick up Ganoderma lucidum spores in a clean bench and inoculate them into solid culture medium. Incubate at 23°C and 90% air humidity.

[0009] (4) After 7-10 days of culture, the mycelium of Ganoderma lucidum grows to the point where obvious colonies can be observed with the naked eye. At this time, pick 10-20 single colonies of Ganoderma lucidum mycelium and transfer them to new PDB solid culture medium respectively, and continue to culture for 7-10 days. After growing again to the point where obvious mycelium can be observed, pick single mycelium colonies and evenly disperse them in 100μl PDB liquid culture medium, and observe the mycelium morphology under a microscope. If the mycelium morphology is intact and there is no contamination from other bacteria or fungi, the single mycelium colony can be used for subsequent fermentation culture. The above steps are all for single mycelium culture of Ganoderma lucidum.

[0010] (5) Pick a single mycelium colony and evenly disperse it in 1 ml of PDB liquid culture medium. Count the number of mycelium under a microscope and adjust the mycelium concentration to 2.5*10 by dilution. 5 / ml.

[0011] (6) PDB liquid medium was used as the fermentation medium. 10% (V / V) mycelium suspension was added to the PDB liquid medium, and 3% (W / V) Schizochytrium algae powder was added at the same time. After being fully stirred, the culture was cultured at 23°C, 90% air humidity, and 200 rpm using a tilting shaker. The mycelium was gently shaken every 4 hours. After 48 hours, microsphere formation was visible: the mycelium self-assembled into microspheres with a diameter of about several hundred microns.

[0012] (7) After fermentation, collect the microspheres, divide the fermentation liquid and the Ganoderma lucidum microspheres into 50 ml centrifuge tubes, and freeze-dry them using a freeze dryer. Wait until the sample is freeze-dried to powder. Dissolve 1 g of the co-fermented freeze-dried powder of Ganoderma lucidum microspheres in 10 ml of PBS buffer to obtain the Ganoderma lucidum microsphere resuspension.

[0013] The present invention is mainly divided into two stages: the cultivation of single mycelium of Ganoderma lucidum and the co-fermentation of Ganoderma lucidum mycelium and algae to form Ganoderma lucidum microspheres. The co-culture and fermentation technology of Ganoderma lucidum microspheres and algae is adopted. First, the Ganoderma lucidum mycelium is inoculated in a special culture medium to promote its growth into a microsphere form. This form is conducive to the efficient exchange of nutrients and the smooth discharge of metabolic waste, thereby improving the efficiency of fermentation and product quality. In addition, the microsphere algae fermentation technology can simulate the symbiotic phenomenon of microorganisms in nature, thereby promoting Ganoderma lucidum to synthesize more beneficial secondary metabolites.

[0014] In a second aspect, based on the above, the present invention provides a co-fermentation product of Ganoderma lucidum microspheres and algae prepared by the above preparation method, which has the effect of repairing hair follicle stem cells and promoting hair growth.

[0015] In a third aspect, based on the above, the present invention provides an application of a co-fermentation product of Ganoderma lucidum microspheres and algae in the preparation of a medicine or health product that has the effect of repairing hair follicle stem cells and promoting hair growth. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 Effects of Ganoderma lucidum microspheres and algae co-fermentation products on hair growth in mice at 15 days under different conditions Figure 2 Effects of different fermentation products on the expression level of growth factors in mouse hair follicle stem cells

Claims

1. A co-fermentation technology of Ganoderma lucidum microspheres and algae that has the function of repairing hair follicle stem cells and promoting hair growth, characterized in that: The steps include: (1) Add potato dextrose medium (PDB) powder into a 1 L conical flask and dilute to 1 L with deionized water. 0.103MPa, 121℃, high pressure steam sterilization for 20-30 minutes, cool to room temperature for subsequent use.

2. (2) Add the PDB culture medium powder to a 1L conical flask, add 10g agarose gel, and use deionized water as the solvent to make the volume 1L. Sterilize with high pressure steam at 0.103MPa and 121℃ for 20-30 minutes. After the culture medium cools to 50-55℃, pour it into a round flat plate culture dish in a clean bench, 20-25ml per dish, and cool to room temperature until solidified.

3. (3) Use sterile tweezers to pick up Ganoderma lucidum spores in a clean bench and inoculate them into solid culture medium. Incubate at 23°C and 90% humidity.

4. (4) After 7-10 days of culture, the mycelium of Ganoderma lucidum grows to the point where colonies can be observed with the naked eye. At this time, pick 10-20 single colonies of Ganoderma lucidum mycelium and transfer them to new PDB solid culture medium respectively. Continue to culture for 7-10 days. After the mycelium grows again to the point where obvious mycelium can be observed, pick single colonies of mycelium and evenly disperse them in 100μl PDB liquid culture medium.

5. (5) Pick a single mycelium colony and evenly disperse it in 1 ml of PDB liquid culture medium. Count the number of mycelium under a microscope and adjust the mycelium concentration to 2.5*10 by dilution. 5 / ml.

6. (6) Using PDB liquid medium as the fermentation medium, add 5-15% (V / V) mycelium suspension to the PDB liquid medium, and add 1-5% (W / V) Schizochytrium algae powder. After fully stirring, use a tilting shaker at 23°C, 90% air humidity, and 200 rpm to culture. Gently shake the mycelium every 4 hours. After 48 hours, microsphere formation can be seen: the mycelium self-assembles into microspheres with a diameter of about several hundred microns.

7. (7) After the fermentation is completed at the corresponding time, collect the microspheres, divide the fermentation liquid and the Ganoderma lucidum microspheres into 50 ml centrifuge tubes, and freeze-dry them using a freeze dryer. Wait until the sample is freeze-dried to powder. Dissolve 1g of the co-fermented freeze-dried powder of Ganoderma lucidum microspheres in 10ml PBS buffer to obtain the Ganoderma lucidum microsphere resuspension.

8. The co-fermentation technology of Ganoderma lucidum microspheres and algae with the function of repairing hair follicle stem cells and promoting hair growth according to claim 1, characterized in that: In step (6), the amount of mycelium suspension added is 10% (V / V), and the amount of Schizochytrium algae powder added is 3% (W / V).

9. The co-fermentation technology of Ganoderma lucidum microspheres and algae with the function of repairing hair follicle stem cells and promoting hair growth according to claim 1, characterized in that: The fermentation time in step (7) is 10 days.

10. The Ganoderma lucidum microsphere extract prepared by the Ganoderma lucidum microsphere and algae co-fermentation technology having the function of repairing hair follicle stem cells and promoting hair growth according to claims 1-3.

11. The medicine prepared from the Ganoderma lucidum microsphere extract prepared by the Ganoderma lucidum microsphere and algae co-fermentation technology having the function of repairing hair follicle stem cells and promoting hair growth according to claim 4.

12. Use of the co-fermentation technology of Ganoderma lucidum microspheres and algae with hair follicle stem cell repairing effect and hair growth promotion according to claim 4 in medicines or health products for promoting hair follicle stem cell repair and hair growth.

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