Lactic acid bacteria having a hair growth and hair nourishing effect

Abstract

To provide proliferation promoters for dermal papilla cells and keratinocytes for the purpose of hair growth and hair restoration effects.SOLUTION: Provided is a lactic acid bacterium belonging to the genus Lactobacillus, the cultures or fermented products thereof, which have the ability to activate dermal papilla cells or promote the production of keratinocyte growth factors. In particular, the lactic acid bacterium is preferably Lactobacillus plantarum N793 strain (NITE BP-03233).SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a strain of lactic acid bacteria of the species Lactobacillus plantarum derived from corn, which has a high ability to activate dermal papilla cells and promote the production of keratinocyte growth factor, and a beverage, food, cosmetic, quasi-drug, and pharmaceutical containing the lactic acid bacteria, its culture solution, or a processed product of its fermentation product.

Background Art

[0002] Thin hair refers to a condition where the activity of hair follicles decreases due to various factors such as aging, stress, lack of sleep, and genetics, resulting in a shortening of the hair cycle. The hair cycle refers to the cycle in which hair repeats three phases: anagen, catagen, and telogen (Non-Patent Document 1). The longest period in the hair cycle is the anagen phase, which is usually about 3 to 6 years. However, due to the factors that shorten the above-mentioned hair cycle, the anagen phase is shortened, causing thin hair.

[0003] The anagen phase is maintained by growth factors such as keratinocyte growth factor, insulin-like growth factor 1, and vascular endothelial growth factor. These growth factors are produced by dermal papilla cells present at the base of hair follicles. Keratinocyte growth factor secreted from dermal papilla cells is known to directly act on hair matrix cells, which are the origin of hair, and promote the proliferation and division of hair matrix cells. Therefore, it is considered to be the growth factor that acts most directly on hair. From the above, dermal papilla cells are necessary cells for hair growth, and if the activation of dermal papilla cells and the increase in the production amount of keratinocyte growth factor can be promoted, it will lead to hair growth (Non-Patent Document 2). On the other hand, Patent Document 1 discloses a method of using an extract of a plant as a promoter for promoting the proliferation of keratinocytes for the purpose of hair growth and hair regrowth.

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Non-Patent Document 2

Patent Document

[0005]

Patent Document 1

[0006] In addition, it is assumed that there are other methods than using the plant extract disclosed in the prior application publication bulletin.

Summary of the Invention

Problems to be Solved by the Invention

[0007] Therefore, the present inventors focused on lactic acid bacteria and utilized the fact that activation of dermal papilla cells and keratinocyte growth factor are essential for hair growth, hair follicle growth, and hair nourishment effects. As a result of screening multiple lactic acid bacteria, they found that there are lactic acid bacteria having high dermal papilla cell activation ability and keratinocyte growth factor production promotion ability, and thus completed the present invention.

Means for Solving the Problems

[0008] That is, the first invention of the present application is a lactic acid bacterium belonging to the genus Lactobacillus, its culture solution or fermentation product having at least one of the abilities to activate dermal papilla cells or promote the production of keratinocyte growth factor. The second invention of the present application is the lactic acid bacterium, its culture solution or fermentation product according to claim 1, which has both the ability to activate dermal papilla cells and the ability to promote the production of keratinocyte growth factor. The third invention of the present application is the lactic acid bacterium according to claim 1 or 2, wherein the lactic acid bacterium belonging to the genus Lactobacillus is Lactobacillus plantarum N793 strain (NITE BP-03233).

[0009] The applicant of the present application also intends food and drink products containing the lactic acid bacterium, its culture solution or fermentation product. That is, the fourth invention of the present application is a food or drink product containing the lactic acid bacterium, its culture solution or fermentation product according to any one of claims 1 to 3.

[0010] Furthermore, the applicant of the present application also intends cosmetics, quasi-drugs, and pharmaceuticals containing the lactic acid bacterium, its culture solution or fermentation product. That is, the fifth invention of the present application is a cosmetic, quasi-drug or pharmaceutical containing the lactic acid bacterium, its culture solution or fermentation product according to any one of claims 1 to 3.

Advantages of the Invention

[0011] The lactic acid bacterium, its culture solution or fermentation product of the present invention has at least one of the ability to activate dermal papilla cells or the ability to promote the production of keratinocyte growth factor.

Brief Description of the Drawings

[0012]

Figure 1

Figure 2

Modes for Carrying Out the Invention

[0013] The present invention will be described in detail below. 1. Lactobacillus plantarum N793 strain (NITE BP-03233) The lactic acid bacteria of the present invention is Lactobacillus plantarum. In particular, among the lactic acid bacteria belonging to Lactobacillus plantarum, it is Lactobacillus plantarum N793 strain (NITE BP-03233). The symbol N793 in the present invention is a number uniquely assigned to the strain by Nissin Food Holdings Co., Ltd., and this Lactobacillus plantarum N793 strain was first isolated by the present inventor. Furthermore, this Lactobacillus plantarum N793 strain has been identified as a lactic acid bacterium belonging to Lactobacillus plantarum subsp. plantarum in more detail.

[0014] The Lactobacillus plantarum N793 strain of the present invention was deposited as NITE BP-03233 with the Patent Microorganisms Depositary, National Institute of Technology and Evaluation on June 18, 2020. The mycological properties of the Lactobacillus plantarum N793 strain of the present invention are as shown in Tables 1 and 2 below. These mycological properties were determined by the method described in Bergey’s manual of systematic bacteriology Vol.2 (1986). Table 1 shows the shape and other characteristics of this strain, and Table 2 shows the results of testing sugar assimilation properties using API 50CH and API CHL (manufactured by bioMérieux). In Table 2, “+” indicates positive and “-” indicates negative.

[0015] [Table 1]

[0016] [Table 2]

[0017] [Table 3]

[0018] 2. Hair papilla cell activation test As shown in the experimental examples described below, Lactobacillus plantarum N793 strain of the present invention has a high ability to activate dermal papilla cells. The confirmation of the ability to activate dermal papilla cells was carried out by the following test method.

[0019] <Preparation of the test substance (lactic acid bacteria powder suspension) used for evaluating the ability to activate dermal papilla cells> The test substance (lactic acid bacteria powder suspension) used for evaluating the ability to activate dermal papilla cells was cultured at 37 °C for 24 hours in MRS medium (manufactured by Becton Dickinson) shown in Table 3 for lactic acid bacteria. Next, the grown bacterial cells were collected using a centrifuge. The collected bacterial cells were washed with sterilized water and collected again using a centrifuge. Washing and collection were repeated three times. After heat sterilization at 95 °C for 15 minutes, collection was performed again using a centrifuge. The collected bacterial cells were freeze-dried using a freeze dryer to obtain dry bacterial cell powder. The dry bacterial cell powder was suspended in PBS (PBS(-) pH 7.4 manufactured by Gibco) to a concentration of 1 mg / mL to obtain a lactic acid bacteria powder suspension.

[0020] <Evaluation of the ability to activate dermal papilla cells> The ability of the test substance obtained above to activate dermal papilla cells was evaluated. That is, human dermal papilla cells were seeded in a 96-well plate at a density of 1.2×10 4 cells / well, and cultured in a CO 2 incubator (5% CO 2 、37 °C) for 6 hours. Then, 100 μL of D-MEM medium (4% FBS) containing each test substance (final concentration 1%) was added. After the addition, it was incubated in a CO 2 incubator (5% CO 2 、37 °C) for 72 hours, and then the number of dermal papilla cells in the culture solution was measured by absorbance photometry. Cell Counting Kit-8 (manufactured by Dojindo Laboratories) was used for the measurement. PBS (PBS(-) pH 7.4 manufactured by Gibco) was used as a control.

[0021] 3. Test for promoting production of keratinocyte growth factor As shown in the experimental examples described below, Lactobacillus plantarum N793 strain of the present invention has a high ability to promote the production of keratinocyte growth factor. The confirmation of the ability to promote the production of keratinocyte growth factor was carried out by the following test method.

[0022] <Evaluation of the ability to promote the production of keratinocyte growth factor> For the same specimens as in the dermal papilla cell activation test, the promoting effect on the production of keratinocyte growth factor was evaluated. In a 48-well plate, human dermal papilla cells were seeded at a density of 1.0×10 4 cells / well and cultured in a CO 2 incubator (5% CO 2 、37°C) for 24 hours. When the culture reached this point, 300 μL of dermal papilla cell medium (manufactured by PromoCell) containing each test specimen group (final concentration 10%) was added. After the addition, the cells were incubated in a CO 2 incubator (5% CO 2 、37°C) for 24 hours, and then the keratinocyte growth factor in the culture supernatant was measured by ELISA. Human KGF / FGF-7 DuoSet ELISA (manufactured by R&D Systems) was used for the measurement. PBS (PBS(-) pH7.4 manufactured by Gibco) was used as a control.

[0023] 4. Bacterial cells, their culture solutions or fermentation products The lactic acid bacteria of the present invention can be used in various forms. For example, if it is the bacterial cells themselves, types in which the bacterial cells are freeze-dried or those obtained by treating the bacterial cells with enzymes can be used. Furthermore, in addition to the bacterial cells themselves, the culture solution of the lactic acid bacteria of the present invention can also be used. Specifically, any of the bacterial cells, the bacterial cells and the culture solution, the culture supernatant, etc. can be used. Also, it goes without saying that the above-mentioned bacterial cells, etc. may be subjected to centrifugation or enzyme treatment with proteases, lipases, etc. as necessary. Furthermore, it may be a fermentation product using the lactic acid bacterium together with the bacterial cells and its culture solution. That is, a fermentation product obtained as a result of fermentation by the lactic acid bacterium using saccharides or the like as a nutrient source can also be used. Specifically, examples of the fermentation product using milk or the like as a raw material include yogurt and cheese. 5. Foods and drinks The lactic acid bacterium, its culture solution or fermentation product of the present invention can be used by being contained in foods and drinks. For example, fermented milk containing lactic acid bacteria and lactic acid bacteria beverages can be considered. In addition, examples of the fermentation product include yogurt and cheese. In the ordinance regarding the ingredient standards of current milk and dairy products, etc., the number of lactic acid bacteria is not particularly specified as an ingredient standard, but for fermented milk (those with 8.0% or more of non-fat milk solids) and lactic acid bacteria beverages (those with 3.0% or more of non-fat milk solids), it is 1.0×10 7 cfu / ml or more, and for lactic acid bacteria beverages (those with less than 3.0% of non-fat milk solids), it is preferably 1.0×10 6 cfu / ml or more, and the above-mentioned bacterial count can be achieved by growing in a fermentation broth such as milk or growing in the form of the final product. In addition to fermented milk containing lactic acid bacteria and lactic acid bacteria beverages, it can also be used for dairy products such as butter, processed egg products such as mayonnaise, confectionery breads such as butter cakes, etc. It can also be suitably used for processed foods such as instant noodles and cookies. In addition to the above, the food of the present invention may be in a formulated form (for example, powder, granule, capsule, tablet, etc.) with appropriate carriers and additives added as necessary together with the lactic acid bacterium, its culture solution or fermentation product of the present invention. Further, the food and drink containing the lactic acid bacterium, its culture solution or fermentation product of the present invention only needs to contain the bacterial cells, its culture solution or fermentation product of the present invention as an active ingredient, and it goes without saying that it may contain other components (for example, saccharides, proteins, amino acids, lipids, moisture, vitamins, minerals, etc.) in addition to the lactic acid bacterium, its culture solution or fermentation product of the present invention.

[0024] The lactic acid bacterium, its culture solution or fermentation product of the present invention is also useful for being contained in foods for specified health use, dietary supplements, etc. in addition to general beverages and foods.

[0025] 6. Cosmetics, quasi-drugs, pharmaceuticals The lactic acid bacteria, culture solution or fermentation product of the present invention can be used by incorporating them into cosmetics, quasi-drugs, and pharmaceuticals in addition to foods. Specifically, hair tonics, hair growth stimulants, hair restorers, and shampoos, rinses, hair lotions, hair tonics, etc. using these can be mentioned. That is, after preparing the lactic acid bacteria, culture solution or fermentation product of the present invention by concentrating, separating, etc. to obtain an extract or powder, it can be used as a cosmetic, quasi-drug, or pharmaceutical containing the said component as an active ingredient. For example, the extract can be packed in a bottle or the like, and the dried fermentation broth powder can be granulated, encapsulated, or tableted using an excipient or the like and provided in the form of tablets. In addition, as cosmetics, quasi-drugs, and pharmaceuticals, it suffices to contain the cells, culture solution or fermentation product of the present invention as an active ingredient, and it goes without saying that other components (for example, saccharides, proteins, amino acids, lipids, moisture, vitamins, minerals, etc.) may be contained in addition to the said component.

[0026] Furthermore, the lactic acid bacteria, culture solution or fermentation product of the present invention can also be applied in the fields of animal feeds and plant fertilizers such as silage, animal feeds, and plant liquid fertilizers.

Industrial Applicability

[0027] The lactic acid bacteria (Lactobacillus plantarum N793 strain), culture solution or fermentation product of the present invention has at least one of high dermal papilla cell activating ability or ability to promote production of keratinocyte growth factor.

Examples

[0028] Examples of the present invention are shown below, but the present invention is not limited to the following examples. <Test Example 1> Evaluation of Dermal Papilla Cell Activating Ability The Lactobacillus plantarum N793 strain of the present invention and three Lactobacillus plantarum comparison strains (N808, N814, N828) owned by the company were evaluated for their dermal papilla cell activating ability.

[0029] The evaluation of the ability to activate dermal papilla cells was performed according to the following procedure. For each of the strain of the present invention and the comparative strain, in the MRS medium (manufactured by Becton Dickinson) shown in Table 3, CO 2 incubator (5% CO 2 , 37°C) for 24 hours. Next, the cells grown using a centrifuge were collected. The collected cells were washed with sterilized water and collected using a centrifuge. Washing and collection were repeated three times. Thereafter, the collected cells were freeze-dried using a freeze dryer to obtain dry cell powder. The dry cell powder was suspended in PBS (PBS(-) pH 7.4 manufactured by Gibco) to a concentration of 1 mg / mL to obtain a lactic acid bacteria suspension.

[0030] Separately, human dermal papilla cells were seeded in a 96-well plate at a density of 1.2×10 4 cells / well, and when cultured in a CO 2 incubator (5% CO 2 , 37°C) for 6 hours, 100 μL of D-MEM medium (FBS 4%) containing each test group (final concentration 1%) was added. After the addition, the cells were incubated in a CO 2 incubator (5% CO 2 , 37°C) for 72 hours, and then the number of dermal papilla cells in the culture solution was measured by absorbance photometry. Cell Counting Kit-8 (manufactured by Dojindo Laboratories) was used for the measurement. PBS (PBS(-) pH 7.4 manufactured by Gibco) was used as a control.

[0031] The activation rates of dermal papilla cells of each strain with the control set to 1 are shown in Table 4 and FIG. 1.

[0032]

Table 4

[0033] As is clear from Table 4 and FIG. 1, it was confirmed that the ability of the lactic acid bacteria (Lactobacillus plantarum N793 strain) of the present invention to activate dermal papilla cells is significantly high and has a high ability to activate dermal papilla cells compared to other lactic acid bacteria owned by the company.

[0034] <Test Example 2> Evaluation of the ability to promote keratinocyte growth factor production The Lactobacillus plantarum N793 strain of the present invention, the reference strain (JCM1149), and three Lactobacillus plantarum comparative strains (N800, N807, N813) owned by the company were evaluated for their ability to promote keratinocyte growth factor production.

[0035] For the same specimens as in the dermal papilla cell activation test, the effect of promoting keratinocyte growth factor production was evaluated. In a 48-well plate, human dermal papilla cells were seeded at a concentration of 1.0×10 4 cells / well, and after culturing in a CO 2 incubator (5% CO 2 24 hours at 37°C), 300 μL of dermal papilla cell medium (manufactured by PromoCell) containing each test specimen group (final concentration 10%) was added. After the addition, the cells were incubated in a CO 2 incubator (5% CO 2 24 hours at 37°C), and then the amount of keratinocyte growth factor produced was measured by ELISA. Human KGF / FGF-7 DuoSet ELISA (manufactured by R&D Systems) was used for the measurement. PBS (PBS(-) pH 7.4 manufactured by Gibco) was used as a control.

[0036] The amounts of keratinocyte growth factor produced by each strain are shown in Table 5 and Figure 2.

[0037]

Table 5

[0038] It was confirmed that the ability of the lactic acid bacterium (Lactobacillus plantarum N793 strain) of the present invention to promote keratinocyte growth factor production was significantly high, and it also had a high ability to promote keratinocyte growth factor production compared to other lactic acid bacteria owned by the company.

Claims

1. Lactobacillus plantarum strain N793 (NITE BP-03233), a lactic acid bacterium having the ability to activate dermal papilla cells and promote the production of keratinocyte growth factor, its culture solution, or fermentation product.

2. Food or drink containing the lactic acid bacterium, its culture solution, or fermentation product according to Claim 1.

3. Cosmetics, quasi-drugs, or pharmaceuticals containing the lactic acid bacterium, its culture solution, or fermentation product according to Claim 1.

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