JPWO2004052323A1 - Rooted sesame extract and hair cosmetic - Google Patents

Abstract

The present invention prevents or improves hair graying by enhancing or restoring the activity of hair follicle melanocytes that have decreased melanin synthesis ability, stopped, or cell death. The problem is to provide an improving agent and a hair growth-promoting agent. Root sesame extract germinated and cultured under temperature shock and / or irradiation with light, comprising anti-heat shock protein (HSP) antibody, anti-fibrocyte growth factor (FGF) antibody and / or anti-melanocyte stimulating hormone (MSH) The problem is solved by incorporating a root sesame extract having reactivity to antibodies into hair cosmetics as an essential component.

Description

  The present invention relates to a hair cosmetic and a pharmaceutical composition or a hair restorer containing a white hair prevention / improving agent for preventing or improving whitening of hair.

The human scalp is exposed to many environmental stresses such as sunlight, ultraviolet rays, temperature changes, chemical substances, and bacterial contamination. Also, anatomically, it is said that the tension of the scalp increases due to the pressure of the cape aponeurosis (gala aponeurotica) on the skull accompanying mental tension, which presses the scalp blood vessels and causes a decrease in blood flow. Yes. In any case, such stress acts suppressively on the cells that make up the scalp and weakens normal functions such as the cells synthesizing proteins to complete their three-dimensional structure. The hair follicle tissue of the human scalp is a complex tissue composed of the outer root sheath, inner root sheath, hair matrix cell, hair follicle melanocyte, hair papilla, sebaceous gland, etc., and is also interpreted as an organ that produces hair. Among them, hair follicle melanocytes hair-full melanocytes are frequently found near the boundary with the hair papilla in the hair bulb hair bulb where the hair matrix cells form a group, and the dendrites of the hair matrix are observed. It extends within the group. In hair follicle melanocytes, melanosomes in the cytoplasm synthesize melanin and supply the melanosomes to hair matrix cells. The hair matrix cells differentiate into hair pulp and fur, thereby completing hair containing melanin. The hair follicle melanocyte repeats the hair follicle melanocyte cycle hair-full melanocycle cycle in conjunction with the hair cycle hair cycle. That is, in the hair cycle regression phase, the hair follicle melanocytes also enter the regression phase, the mature melanosomes disappear, and only contain a few immature melanosomes. In the rest period of the hair cycle, the base of the hair is keratinized and swells in a club shape. At this time, hair matrix cells, hair follicle melanocytes, and hair papilla disappear.
The color of the hair is the maturity of the melanosomes in the follicular melanocytes, the type, quality and amount of melanin synthesized by the melanosomes, the degree of migration of the melanosomes into the hair matrix cells, the fur cells and their keratinized layers. It depends on the distribution density, morphology, presence or absence of disintegration of melanosomes. Melanin synthesized by melanosomes is roughly classified into black eumelanin (also called true melanin) and yellow-brown pheomelanin (also called pseudo melanin). In black hair, eumelanin is mainly used, in blonde blond, pheomelanin is mainly used, and in brown hair, both melanins are mixed.
In Japanese black hair, hair follicle melanocytes coexisting with hair matrix cells synthesize melanin (exactly eumelanin) and supply it to hair matrix cells, which contain melanin. Complete by differentiating into fur cells. About white hair that occurs with aging, general interest is high along with thinning hair and wrinkles, but there are many unclear points about its origin. At the cellular level, gray hair is caused by a decrease in the function of hair follicle melanocytes existing in the hair root part or cell death (apoptosis), and melanin supply is stopped, and stress and aging are thought to be the cause. From the viewpoint of cell dynamics, hair follicle melanocytes repeat the melanocyte cycle in conjunction with the hair follicle hair cycle, but the vitality of hair follicle melanocytes is weaker than that of hair matrix cells, which is due to stress, aging and other factors. It is easily damaged. In electron microscopic studies on gray hair, hair follicle melanocytes were not completely killed, but the appearance of vacuoles in the cytoplasm and a marked decrease in melanosomes were observed, suggesting their functional decline.
Attempts to prevent white hair are considered more difficult than hair growth, and there are few anti-white hair products compared to hair restorers. Many current gray hair prevention / improving agents describe the effectiveness of random screening and specified substances using melanocyte enhancement of melanin production or tyrosinase activity as an index. For example, a benzosophenone derivative is used to increase melanin production and promote tyrosinase activity in cultured melanoma cells (Patent Document 1), and there is a chemical substance that increases cyclic AMP having various cell function enhancement actions. Those using this to activate melanocytes (Patent Document 2), those using cyclic AMP or its derivatives to activate hair root melanocytes (Patent Document 3), and specific chromone derivatives to hair follicles Utilizing this to cause activation of melanocytes (Patent Document 4), using lignin isolated from hardwood wood to promote tyrosinase activity of melanocytes (Patent Document 5), vasoconstrictive action Endothelin with cell proliferation increases melanocytes And those utilizing this as causing melanin production increase (Patent Document 6), adenosine cyclic phosphate compounds and forskolin or derivatives thereof utilizing this as causing melanocyte activation (Patent Document 7), tropical plants Copaiba extract used to promote melanin production in cultured melanoma cells (Patent Document 8), adenosine phosphate compounds (AMP, ADP, ATP) cause proliferation of hair follicle melanocytes and increased tyrosinase activity (Patent document 9) etc. which use this as these are mentioned, However, the gray hair improvement effect is not confirmed in a practical use.
By the way, when a biological tissue cell is exposed to a temperature 5 to 10 ° C. higher than normal temperature, it is generated in the cell and has a function of protecting the cell (heat-shock protein, hereinafter referred to as “HSP”). ) Are a group of proteins whose synthesis is induced at high temperatures, but are also called stress proteins because they are induced by various environmental stresses other than high temperatures. Environmental stress includes physical changes in temperature, ultraviolet rays, radiation, atmospheric pressure, and other chemical substances, such as chemicals, drugs, poisons, and metal ions. Blood, nutrient deficiency, hypoxia, active oxygen and the like can be mentioned. HSPs are thought to be involved in biological defense mechanisms that respond to these adverse environmental conditions.
The main molecular biological function of HSP is to bind to a new protein that has just been synthesized in the cell and has not yet taken a three-dimensional structure, and prevents the modification due to the formation of the intermolecular association to complete the correct three-dimensional structure. It is a so-called chaperone function that facilitates the degradation and regeneration of denatured proteins.
HSP is called with a molecular weight, and molecular weights of 90,000, 70,000, 60,000 (mass is 90 kDa, 70 kDa, 60 kDa, respectively) are well known. It is called HSP90, HSP70, HSP60, etc. On the other hand, there are also small HSPs having a molecular weight of 50,000 or less, which are called small HSPs (sHSPs), and have recently attracted attention for their functions to avoid cell death (apoptosis) and to support the differentiation of young cells. Yes. Although sHSP is not so much in animal cells and yeast, it is considered to exist in plant cells. In general, HSP exists widely in the living world from bacteria to higher animals and plants, and has high homology. Since this protein is an essential molecule for cell survival, it is thought that it has been well preserved across species in the history of biological evolution. Among the HSPs, the functions of HSP27 having a molecular weight of 27,000 and HSP25 having a molecular weight of 25,000 are functions that restore the functional degradation of protein synthesis by apoptosis-suppressing function (Non-patent Document 1, etc.) and chaperone function (non-patented) Reference 2), function of inhibiting necrosis of tumor necrosis factor TNF (Non-patent Documents 3 and 4, etc.), function of supporting the differentiation of juvenile cells (Non-patent Document 5), fertilization of fertilized eggs on endometrium There are reports of functions that support floor and growth, and placental maintenance (Non-Patent Document 6). However, it is not known that HSP27, HSP25, etc. have the effect of preventing white hair improvement and hair growth.
On the other hand, along with white hair, there is a great demand for hair growth agents that combat thinning hair and alopecia. The classification of alopecia is as follows: 1) Physiological alopecia (male pattern alopecia male pattern alopecia or androgenetic alopecia, alopecia with age, etc.), 2) congenital alopecia, 3) acquired Alopecia (alopecia areata alopecia, alopecia associated with skin diseases, etc.) is classified broadly, and those targeted for hair growth agents are physiological alopecia and acquired alopecia. Analyzing the mechanism of physiological or acquired alopecia from a tissue or cell perspective, hair loss is a decrease in the activity of the root sheath cells that maintain the hair during the hair cycle resting period of the scalp hair follicle, and the resulting loss of hair loss. It occurs due to an increase, a decrease in the activity of hair matrix cells or hair papilla cells in the growing hair follicle, or a decrease in hair production ability as a result. In order to maintain a sufficient number of hairs, the root sheath cells of the resting hair follicles remain active to prevent hair loss and the hair follicle hair matrix cells and the dermal papilla cells that support them remain active. Therefore, it is necessary to synthesize hair constantly, maintain the activity of capillaries of hair papilla and connective woven follicles, and maintain sufficient blood circulation. The active ingredients of various commercially available hair restorers can be classified into 1) activation of hair follicle cells, 2) suppression of male hormones, 3) promotion of blood circulation of the scalp, 4) general care of the scalp, and the like. 1) Targeting hair follicle cell activation is pentadecanoic acid glyceride, procyanidin, etc. 2) Targeting male hormone suppression is aromatase, diethylstilbesterol, dipotassium glycyrrhizinate, etc. 3) Blood circulation Targets for promotion include minoxidil, vitamin E, niacin, carpronium chloride, garlic extract, etc. 4) Targets for scalp care include aloe extract, placenta extract, hinokitiol, amino acids, etc. We are calling for effectiveness.
On the other hand, as hair cosmetics using germinated sesame seeds, lignan glycosides are extracted with hydrated lower alcohols (such as methyl alcohol) from sesame seeds cultured and germinated at 37 ° C. (Patent Document 10). Alternatively, there is a hair cosmetic effect obtained by combining with a cell activator (Patent Document 11), but physical stimuli such as light and temperature changes applied during sesame culture improve white hair in the sesame extract. A substance having reactivity with one or more antibodies selected from the group consisting of anti-heat shock protein (HSP) antibody, anti-fibrocyte growth factor (FGF) antibody and anti-melanocyte stimulating hormone (MSH) antibody There is no description or suggestion of inducing. In addition, a topical skin preparation that combines a lignan glycoside obtained by hydrous alcohol extraction from germinated sesame and a tyrosinase activity inhibitor is disclosed (Patent Documents 12 and 13). Has an inhibitory effect on tyrosinase activity alone, and the above-mentioned external preparation for skin is expected to have a synergistic whitening effect in combination with other tyrosinase activity inhibitors, and it is necessary to activate tyrosinase. It has an effect that is completely opposite to the effect of improving and preventing white hair.

The present invention relates to a hair follicle melanocyte that has been reduced in ability to synthesize melanin, or an agent for improving or preventing white hair that enhances the activity of melanin producing cells in the skin or tissue and restores the ability to synthesize melanin, thereby preventing or improving whitening of hair. Or it makes it a subject to provide a hair restorer.
The present inventors focused on heat shock protein (HSP), fibroblast growth factor (FGF) and melanocyte stimulating factor (MSH). According to an immunohistochemical study on human scalp tissue, a substance that reacts with an antibody reactive with HSP25 having a molecular weight of 25,000 and HSP27 having a molecular weight of 27,000, which is sHSP, of HSPs, A substance having a binding property to an antibody (anti-HSP antibody) that reacts with HSP25 and HSP27; It has been found that the sesame new root extract contains an effect of improving white hair prevention and promoting hair growth. Furthermore, through further immunohistochemical studies, substances that bind to antibodies reactive with cell growth factors FGF2 and FGF4 (anti-FGF antibodies) will be present in human scalp hair bulb cells and root sheath cells. In order to produce substances that bind to these specific antibodies, it is necessary to root sesame under specific conditions such as applying a temperature shock, and this rooted sesame extract has a hair-growth effect. I found out.
In addition to the specific conditions described above, some cells of sesame new roots (straight roots) cultured under light irradiation produce substances that are reactive with melanocyte-stimulating hormone antibodies (anti-MSH antibodies). Made a new discovery that a number of cells produce melanin. Furthermore, the extract of root sesame cultivated under these conditions reacts with the above-mentioned anti-HSP antibody and anti-FGF antibody, and causes remelanization of white hair, thereby having an effect of improving white hair prevention and promoting hair growth. I found out.
Here, a supplementary explanation will be given of the opportunity for the discovery of the substance having the anti-MSH antibody reactivity. The inventors of the present application initially conducted research for the purpose of searching for substances having reactivity with anti-HSP antibodies and anti-FGF antibodies. In sesame culture, in order to avoid the involvement of shock factors other than heat shock, the sesame in the culture was kept under the condition that the window curtain was closed so that sunlight from the window did not hit. However, one night, I forgot to close the window curtain, so I failed to see the direct sunlight from the Asahi hit the root sesame in culture. When the new roots of the sesame were closely observed, the pure white new roots appeared to be slightly yellowed (see Example 6 and FIG. 11). I interpreted this faint discoloration as an injury caused by direct sunlight and tried to dispose of this rooted sesame, but I decided not to do so and decided to elucidate the true nature of this yellowing. Immediately, the rooted sesame was fixed in formalin and embedded in paraffin to prepare a tissue section. I thought that the coloration of sesame new root was due to melanin, but I was prepared for the disappointment, but when the tissue section of this sesame new root was stained with melanin by the Fontana silver method, the cytoplasm of the epidermis cells of sesame new root turned black It was found to be stained and melanin positive (see Example 7 and FIG. 13).
For reconfirmation, sesame seeds were cultivated in a place exposed to sunlight from the beginning, but the microscopic findings of sesame new roots were the same. When comparing black and white sesame seeds, the amount of melanin was higher in white sesame seeds under microscopic findings. When sesame was cultured with light blocked by cardboard or the like, the amount of melanin was clearly reduced (see FIG. 15). Visual observation of sesame seedlings that were exposed to direct sunlight revealed that this coloring occurred at the root of the new roots.
The production of melanin in rooted sesame epidermal cells suggests that melanocyte-stimulating hormone-producing cells exist somewhere in the new root tissue. The inventor of the present application thinks that anti-alpha / melanocyte-stimulating hormone antibody, which is an antibody against alpha-melanocyte stimulating hormone (hereinafter referred to as α-MSH), with respect to these tissue sections of sesame new roots. (Histochemistry) using immunohistochemical staining (hereinafter referred to as immunostaining), a cell with cytoplasm stained positive for α-MSH antibody was found inside the sesame new root. (See Example 8 and FIG. 14).
Next, in order to confirm melanin and α-MSH-secreting cells with a transmission electron microscope, sesame new roots rooted in direct sunlight were observed by a normal electron microscopic method. A high electron density substance that appears to be melanin was observed in the cytoplasm and along the nuclear membrane in the nearby epidermal cells (see Example 9 and FIG. 16a). When the sesame new root portion in which α-MSH antibody-positive cells were previously observed by light microscope observation was observed by an electron microscopic method, cells containing abundant Golgi apparatus and secretory granules were found (Example 9 and FIG. 16b).
Here, the sesame new root tissue is explained in terms of plant anatomy. When the sesame absorbs water sufficiently and rooting begins, the radicle first protrudes from the hypocotyl, breaks the shell, and extends outside to become a new root. . The new root is also called a direct root taroot (or main root main root), and its tissue structure is epidermis epidermis as the outermost layer, cortex cortex as the inner layer, and endodermis as the innermost layer of the cortex. A portion from the inner side of the cortex to the center of the root is a central pillar steel (central cylinder). In the central column, an inner sheath pericle is formed on the surface layer, and a vascular bundle is formed on the inner side. The vascular bundle is formed of a xylem that is a path for water and inorganic nutrients, a sieve phloem that is a path for organic nutrients, and a parenchyma cell that fills the space between them (see FIG. 12).
When immunostaining specimens for melanin and α-MSH are examined based on these structures, melanin-positive cells are epidermal cells, and α-MSH-positive cells are thought to be endothelial cells (or inner sheath cells). . It is considered that α-MSH newly discovered in sesame new root cells in the present invention is produced inside sesame, which acts on epidermal cells of new roots to produce melanin. The central root of the new root is an important juvenile that is the pre-formed layer procambium involved in the formation of vascular bundles, which can be called the lifeline of the new root, and the lateral meristem of root that causes the elongation and differentiation of the roots. Includes cell populations. Melanin formed in the epidermis is thought to protect these immature cell groups in the central column from light injury. And the extract extracted from this root sesame was found to have the effect of reacting with anti-HSP and anti-FGF antibodies and blackening gray hair by reacting with anti-MSH antibodies.
Here is a supplementary note on melanocyte stimulating hormone (MSH).
Melanin is a polymer pigment that exists not only in higher organisms such as animals and plants but also in the living world such as microorganisms. It is synthesized from melanocytes and helps absorb excess light. In the human body, melanocytes are not limited to the skin that is exposed to light, but have long been confirmed throughout the body, such as the oral cavity, pharynx, sinuses, and esophagus that are not normally exposed to light (Non-Patent Document 7). . Regarding the function of melanocytes at such sites, for example, in the case of oral mucosa, when melanocytes cause melanin production even when hot things that are likely to burn in the mouth, melanin is light or In addition to being a substance that blocks ultraviolet rays, it is considered to be one of biological substances that act to alleviate or recover from tissue damage due to various stresses and damaging factors applied locally.
What stimulates melanocytes to cause melanin synthesis is a peptide hormone (cytokine) called melanocyte-stimulating hormone (MSH), and there are three forms of α, β and γ. Recent research has revealed that MSH and its related substances have various other physiological functions as described below.
Among them, α-MSH is a linear polypeptide composed of 13 amino acid residues having no difference in animal species, the same primary structure, and acetylated N-terminal ser. Reports where α-MSH was found in plants containing sesame were not found in the searched range. In vertebrates, MSH is produced by intracellular processing from propiomelanocortin (hereinafter referred to as POMC), which is a common precursor protein such as ACTH (adrenocorticotropic hormone corticotropin) and β-endorphin. It is known that The mammalian POMC gene is expressed in various tissues such as skin, gastrointestinal tract, lung, kidney, placenta, testis, and ovary, and is thought to be paracrine and exert various functions in local cells. (Non-Patent Document 8). There has been a report of detecting POMC in trace amounts in testis, duodenum, kidney, colon, liver, lung, stomach, spleen and the like, which are tissues other than the pituitary gland, by gel filtration chromatography using an antibody (Non-patent Document 9). ).
Further, MSH and ACTH are sometimes collectively referred to as melanocortin, because most of the MSH action is common with the action of ACTH, and some of them contain a common amino acid sequence. In recent years, receptors (receptors) of MSH or melanocortin are not limited to melanocytes, but have begun to be found in cells of various parts of the human body, and their functions and methods of use for medical care or health maintenance are attracting attention.
For example, since it is expressed in macrophages, it is used as an anti-inflammatory peptide (Non-patent Document 10), and it is also expressed in adipose tissue. Actions involved in higher cranial nerve activities such as learning, memory, vision, hearing and whole body from the expression in the hippocampus, thalamus, hypothalamus, brain stem and cerebral cortex (Non-patent Document 12), and α- Since MSH suppressed tumor necrosis factor-α-TNF (tumor necrosis factor-alpha) in the mouse brain, the brain protection action against cerebral ischemia (Non-patent Document 13), from the protective action of transplanted heart to transplanted medicine PC1, which is one of the processing enzymes that make melanocortin by intracellular processing from POMC by intracellular processing (Non-Patent Document 14), Diabetes is suppressed from being localized in insulin cells (β cells) in Ngerhans Island (Non-patent Document 15), and by using both α-MSH and TGFβ2 (transforming growth factor-beta 2) in combination, experimental autoimmunity Have been reported to be used for autoimmune inflammatory diseases (Non-patent Document 16) because of their ability to suppress uveoretinitis, and endocrine diseases such as inflammation treatment, obesity suppression, brain activity support, diabetes, etc. It can also be used in a wide range of medical areas such as stroke, organ transplantation, and autoimmune diseases, and in pharmaceutical compositions and foods and drinks that address health maintenance. In this way, various effects can be obtained by controlling the action of MSH.
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Non-Patent Document 17: Book edited by Coperand LO et al. “Seed science and technology” Kluwer Academic Publishers Boston Dorderecht London 2001, page 71 The present invention was made in this background. The configuration is as follows.
(1) Reactivity to one or more antibodies in the group consisting of anti-heat shock protein (HSP) antibody, anti-fibrocyte growth factor (FGF) antibody, anti-melanocyte stimulating hormone (MSH) antibody Rooting sesame extract characterized by
(2) The root sesame extract according to (1), wherein the anti-heat shock protein (HSP) is HSP25 and / or HSP27.
(3) The root sesame extract according to (1) or (2), wherein the anti-fibrocyte growth factor (FGF) is FGF2 and / or FGF4.
(4) The root sesame extract according to any one of (1) to (3), wherein the anti-melanocyte stimulating hormone (MSH) is α-MSH.
(5) Root sesame extract having reactivity to any one or more of the group consisting of HSP25 and / or HSP27 antibody, FGF2 and / or FGF4 antibody, α-MSH antibody .
(6) A cosmetic for hair comprising the rooted sesame extract according to (1) to (5).
(7) An agent for preventing and / or improving gray hair comprising the rooted sesame extract according to any one of (1) to (5).
(8) A pharmaceutical composition comprising the rooted sesame extract according to (1) to (5).
(9) A hair growth promoter comprising the rooted sesame extract according to (1) to (5).
(10) A food or drink comprising the rooted sesame extract according to (1) to (5).
In this specification, “rooting sesame” is used in the following definition.
The phenomenon generally called germination, sprouting, seedling is defined as an emergence of the radical through the seed coat (non-patent document 17). Therefore, germination may be rooted. However, since the word rooting is not common, in the present specification, rooting (germination) and both are sometimes described later. In the case of sesame seeds, the new roots that have started to break through the seed shell of sesame seeds are sometimes called straight roots (or primary roots).

FIG. 1 is a photomicrograph of the hair root tissue observed in an HSP27 immunostained specimen of human scalp. (200 times). The cytoplasm of the hair matrix is positive for Hsp and is shown dark in this black and white photograph (arrow).
FIG. 2 is a photomicrograph of hair root tissue observed in an FGF-2 immunostained specimen of human scalp. (400 times). The cytoplasm of dermal papilla cells is FGF-2 positive and is shown in dark shade in this black and white photograph (arrow).
FIG. 3 is a photomicrograph of decidual cells observed in an HSP27 immunostained specimen of human placenta. (400 times). The cytoplasm of the shed cells is Hsp27 positive and is shown in dark shade in this black and white photograph (arrow).
FIG. 4 is a photomicrograph of chorionic vegetative cells found in human placental HSP27 immunostained specimens. (400 times). The cytoplasm of cytotrophoblast (arrow) and syncytiotrophoblast (arrowhead) of cytotrophoblasts (arrowheads) is Hsp27 positive and is shown in dark tone in this black and white photograph.
FIG. 5 is a photomicrograph of a new root tip tissue observed in an HSP27 immunostained sample of rooting sesame. (200 times). The cytoplasm of surface cells at the tip of the new root is Hsp27 positive, and is shown in dark tone in this black and white photograph (arrow).
FIG. 6 is a photomicrograph of the tip tissue of a new root found in a MIB1 immunostained specimen of rooting sesame. (200 times). The nuclei of proliferating cells at the tip and medulla of the new root show MIB1 positivity and are shown in dark shade in this black and white photograph (arrows).
FIG. 7 is a photomicrograph of the tip tissue of a new root found in an FGF-2 immunostained specimen of rooting sesame.
(100 times). The cytoplasm of the surface layer and the medullary cell at the tip of the new root is FGF-2 positive, and is shown in darkness in this black and white photograph (arrow).
FIG. 8 is a photomicrograph of a new root tip tissue observed in an FGF-2 immunostained specimen of rooting sesame. (50 times). The dendritic structure corresponding to the primitive veins is FGF-2 positive and is shown in dark shade in this black and white photograph (arrow).
FIG. 9 shows a micrograph of a specimen that is blackened by melanin staining by the Fontana silver method at a portion blackened by root sesame extract / lotion of human hair (100 times).
FIG. 10 shows a photomicrograph of the tip of the hair whose root is blackened by the rooting sesame extract / lotion of human hair (100 times). The tip is not blackened and remains gray.
FIG. 11 is an enlarged photograph of rooted sesame. a: Rooted sesame seeds cultured while exposed to light during the day. b: Rooted sesame seeds cultivated by applying light during the day and applying a low temperature shock at night. c: Rooted sesame seeds cultured in the daytime by blocking light and giving a low temperature shock at night. In rooting sesame seedlings cultured under light, a pale yellow-brown color is observed at the base of new roots (arrows), but in rooting sesame seeds cultured after blocking light, they are white (arrowheads). . (The scale is 1 mm per scale)
FIG. 12 is a schematic diagram showing the tissue structure of rooted sesame.
FIG. 13 is a photomicrograph of a melanin-stained specimen of a rooted sesame cultured under irradiation of light using the Masson-Fontana method. Black positive findings are observed in the epidermis cells of the light yellowish brown part of the new root of sesame (the part surrounded by a square). The shooting magnification is 100 times.
b: An enlarged photograph of cells that are positive for melanin staining. Black silver particles generated by the Masson-Fontana method are observed not only in the surface cells but also in the second and third layer cells (the part enclosed by a square). The shooting magnification is 400 times.
FIG. 14 is a photomicrograph of an immunostained specimen using an anti-α-melanocyte-stimulating hormone (α-MSH) antibody of rooting sesame cultured under light irradiation. The cytoplasm of one layer of the cortical deep layer of the sesame new root base is colored brown (black in the photograph), and α-MSH positive findings are observed (the part surrounded by a square). The shooting magnification is 100 times. b: Magnified photograph of positive cells in an immunostained specimen using rooted sesame anti-α-MSH antibody. The cytoplasm of the deepest cortical cell is colored brown (black in the photograph), and α-MSH positive findings are observed (the part enclosed by a square). The shooting magnification is 400 times.
FIG. 15 is a photomicrograph of a melanin-stained specimen of the rooted sesame cultured with the light blocked by a Masson-Fontana method. At the base of sesame new root, only a slight black positive finding is observed in the epidermal cells (the part enclosed by a square). The shooting magnification is 100 times. b: An enlarged photograph of cells of a melanin-stained specimen. A small number of black silver particles generated by the Masson-Fontana method are observed around the nucleus of the surface cell (the portion surrounded by a square), but are hardly observed in the cells of the second and third layers. The shooting magnification is 400 times.
FIG. 16 is a transmission electron micrograph of rooting sesame cultured while being irradiated with light. a: Many fine granules with a high electron density that are thought to be melanin are observed in the cytoplasm of epidermal cells. In addition, the deposition of a uniform and non-structured high electron density material is observed along the nuclear membrane. b: A part of the cytoplasm of the deepest cortical cell of the new root base. The Golgi apparatus (Δ) is remarkably developed, and a ring structure (↓) with a high electron density that seems to be a secretion is observed in the vicinity. Photo magnification is 34,500 times.
c: shows an α-MSH immunoelectron micrograph of the cortical deepest cell of the new root base. In the cytoplasm, a large number of small granules having a high electron density of 120 to 150 nm (one of which is indicated by an arrow) were identified, and these small granules were positive for α-MSH, and were determined to be α-MSH granules (Fig. 16c). The photo magnification is 34,500 times.
FIG. 17 is an enlarged photograph of an eyebrow sample that was extracted after applying an extract of rooted sesame seeds cultivated while irradiating light to the eyebrows including white hair for two months. One with white hair (a) and one with white hair but the roots are black (b) are shown. Those with white hair are those in the rest period of the hair cycle from the point that the root ends are swollen in a rod shape. The blackened part of the hair is from the hair cycle growth stage because the root of the hair is swollen as if the cup is turned upside down.
FIG. 18 shows the variation in the extraction amount of the substance that reacts with the anti-MSH antibody by changing the extraction solvent.
FIG. 19 shows changes in the extraction amount of a substance that reacts with an anti-MSH antibody due to a change in acetic acid concentration when acetic acid or cold water is used as an extraction solvent.
FIG. 20 shows the results of examining the effect of water quality on rooting conditions.

1. Preparation of root sesame extract having reactivity to anti-HSP antibody and anti-FGF antibody
As described above, ultraviolet rays and other various stresses are applied to the human scalp to adversely affect hair follicle cells. In order to protect hair matrix cells and hair follicle melanocytes from this stress, it is possible that heat shock proteins (stress proteins) are produced in the hair follicle as a biological defense mechanism. Although there are many types of heat shock proteins, it is known that female hormones are related to hair growth. Therefore, among heat shock proteins, HSP25 which is a heat shock protein particularly related to female hormones and As a result of examining and examining HSP27, it was found immunohistochemically that substances reacting with anti-HSP27 and anti-HSP25 antibodies are present in cells of human scalp hair bulbs. (See Experimental Example 1)
Furthermore, in order to know the relationship between anti-HSP25 and anti-HSP27 antibodies and female hormones, human placental tissues were examined for their reactivity with anti-HSP25 and anti-HSP27 antibodies. And immunohistochemically found to be present in the vesicles of trophoblast trophoblast cells of placenta. (See Experimental Example 2).
On the other hand, from the viewpoint of the raw material of hair cosmetics, attention is given to sesame, which has been said to be good for hair from a long time ago, and a substance that reacts with anti-HSP25 and anti-HSP27 antibodies by rooting sesame under the stress of temperature fluctuation Was found to be produced (see Example 1).
The fact that substances reacting with anti-HSP25 or anti-HSP27 antibodies have been proved in sesame new root (first appearing root) cells protects young cells of sesame new root from stress and synthesizes protein as a chaperone. It seems to be acting on cell death (apoptosis) prevention.
Substances that react with the anti-HSP25 and anti-HSP27 antibodies can be obtained by rooting sesame seeds while giving a temperature difference and obtaining them from the new root cells of the sesame seeds. By supplementing the scalp with an extract containing an anti-HSP25 or anti-HSP27 antibody-reactive substance obtained by extraction from sesame new roots, melanin synthesis is weakened or stopped, or cell death tends to occur. The activity of hair follicle melanocytes can be restored and white hair can be prevented and improved. That is, HSP25 or HSP27, which is a stress avoidance component possessed by plant cells, can be used in humans to provide a hair cosmetic excellent in activation of hair matrix cells and hair follicle melanocyte cells.
HSP25 or HSP27 recovers the activity of hair follicle melanocytes that have a tendency to decrease, stop, or cause cell death, and prevent and improve whitening of hair. Therefore, by applying HSP25 or HSP27 to the scalp having gray hair, blackening of gray hair can be induced.
Since the substance that reacts with the anti-HSP25 or anti-HSP27 antibody is a protein, a solvent that coagulates the protein is not suitable for use as a cosmetic. Substances that react with anti-HSP25 and anti-HSP27 antibodies are prone to degradation and denaturation, and therefore, such means as protection by microcapsules or liposomes, freeze drying, etc. are used for cosmetics. Can be provided as stable.
The sesame used in the present invention is not limited as long as the cells are alive. Specifically, raw sesame (seed sesame) is used. Those that have been subjected to treatment such as sun drying and fumigation cannot be used because their cells are dead.
In order to produce the root sesame used in the present invention, appropriate consideration is required for temperature management, water supply, ventilation, daylighting, and the like. Select a condition that approximates the rooting situation in the natural world where the roots are stretched in the difference between the warmth and the coldness of the day when seed sesame is planted in the field in early spring, which is warm during the day and cool at night. It is preferable. In general, heat shock proteins are produced when cells are exposed to a temperature difference of 10 ° C. for several hours. Therefore, this temperature difference may be used to produce the root sesame seeds of the present invention. Conditions such as water supply and ventilation may be set as appropriate according to the seed sesame used. In general, it is sufficient that the water content is such that sesame seeds do not dry. When water is used, water is appropriately changed so that the water does not change in quality, but mist or water vapor can be used instead of water. The water to be used may be tap water, but if there is a problem with the quality of tap water, it is better to use groundwater, and various mineral water, deep ocean water, hot spring water, etc. may be used. Since the oxygen demand of rooted sesame increases with the passage of time, it is desirable to supply oxygen corresponding to the time and the density of sesame.
In addition, since sesamin, which is widely known as a component of sesame, has disappeared on the 2nd to 3rd days from the root of sesame, the root sesame in the present invention cannot be expected. As for sesame oil, when rooting sesame is observed with an electron microscope, a small number of fine lipid droplets may be observed in the cytoplasm of some cells, but these are not sesame oil, because they are fine. Pathologically, it is interpreted as steatosis based on nutritional deficiency and oxygen deficiency.
When sesame roots and new roots reach 2 to 3 mm, stop culturing and be careful not to damage the new roots to make new root homogenates, but the new roots are easily damaged, and if they are damaged Since active ingredients such as HSP are easily lost from the cells, the whole rooted sesame may be homogenized as it is without separating new roots in order to avoid it. In the immunohistochemical study conducted this time, substances that react with anti-FGF2 and anti-FGF4 antibodies in cells other than new roots were also found in tissues other than new roots, so it is preferable to homogenize the whole rooted sesame. When rooting sesame is homogenized, it is necessary to take measures to prevent degradation and denaturation of the active ingredient. As an example of the treatment, it is preferable to complete the homogenate operation in a short time as possible at a low temperature, and it is preferable to use water or another neutral solvent (for example, 50 mM Tris-HCl buffer) as the extraction solvent for the active ingredient. When using an alcohol or other organic solvent, it is necessary to adjust the concentration so as not to alter the active ingredient. The homogenate is filtered or centrifuged, and the filtrate or supernatant is used as a rooted sesame extract, but may be called a germinated sesame extract. This extract can be used as a raw material in hair cosmetics. Since the root sesame extract is easily degraded and decomposed, the stock solution is preferably stored frozen. Usually, such a rooted sesame extract contains 0.01 pg to 100 μg of a substance that reacts with anti-HSP25, anti-HSP27, anti-FGF2 or anti-FGF4 antibody per ml of the extract.
A rooting sesame extract containing a substance that reacts with anti-HSP25, anti-HSP27, anti-FGF2 or anti-FGF4 antibody can be provided as a hair cosmetic. When rooting sesame extract is blended into hair cosmetics, the blending amount differs depending on the rooting state, extraction method, extraction efficiency, active ingredient stabilization treatment method, etc. Since it differs depending on the dosage form, it can be appropriately set depending on various circumstances. For example, when a root sesame extract having approximately the same mass as the wet mass of root sesame is produced, the same extract can be blended into the hair cosmetic at a ratio of 10 to 30% by mass.
For hair cosmetics, oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, sequestering metal ions Agents, polymers such as water-soluble polymers, thickeners, powder components, UV absorbers, UV blockers, perfumes, pH adjusters, and the like. In addition, other medicinal components such as vitamins, skin activators, blood circulation promoters, active oxygen scavengers, anti-inflammatory agents, and bactericides, and physiologically active components can be contained.
The composition containing the root sesame extract of the present invention can be applied in the form of a semi-solid agent such as an aqueous solution, an oil agent, a milky lotion, a liquid solution such as a soap solution, a gel, or a cream. It can be prepared in these forms by a conventionally known method to form various dosage forms such as lotions, emulsions, gels, creams and aerosols. These can be applied to the scalp by spraying or spraying. Of these dosage forms, lotions, emulsions, creams, aerosols and the like are particularly suitable. Usually, these dosage forms can be produced according to the formulation method used in cosmetics.
Since sesame is originally a food, rooted sesame can be used as a food or supplement as well as bean sprouts made from soybeans. At that time, it is preferable to prevent the physiological activity of the active ingredient from being lowered by performing a treatment such as freeze drying.
As food, it is eaten as a health food or food material useful for prevention or treatment of gray hair, as it is, or by adding various nutritional components or adding it to foods and drinks. After adding suitable starch, vegetable oil and other auxiliaries, it can be used for edible by using conventional means to form it into an edible form, such as granules, granules, tablets, capsules, pastes, etc. In addition, it may be used by adding to various foods such as bread and fermented dairy products, or by adding to beverages such as soft drinks.
2. Production of root sesame extract reactive with anti-MSH antibody
The inventors of the present application have found anti-α-MSH antibody positive cells in the new roots by immunohistochemical study of cultured sesame. When rooted sesame was homogenized to produce a rooted sesame extract and applied to the scalp and eyebrows of a male with gray hair, a strong white hair improvement effect was confirmed (see Example 6, FIG. 7).
“Sesame culturing performed while irradiating light” in Example 6 is a stage in which new roots emerge from the shell from cultivated sesame, and light rays such as sunlight, ultraviolet rays, various artificial acupuncture rays It refers to hitting. As described above, new roots contain many undifferentiated immature cells that are involved in root elongation and differentiation, and these are susceptible to photodamage. Therefore, the type, intensity, and irradiation time schedule for appropriately producing a substance that reacts with the anti-α-MSH antibody in sesame new roots must be determined while observing the growth status of sesame new roots. (See Example 7)
The light intensity applied to the root sesame may be determined in the range of 500 to 50,000 lux. (The brightness of ordinary lighting in the room is 1,000 to 2,000 lux, and the sunlight is 10,000 to 100,000 lux). Of course, even if it is a production condition which does not require light irradiation, if it is a root sesame extract cultivated and prepared on the conditions containing the substance which reacts with an anti-alpha-MSH antibody, it will be included by this invention. In the present specification, a substance that reacts with an anti-α-MSH antibody may be described as α-MSH.
The sesame extract used to obtain a root sesame extract containing a component that reacts with the anti-α-MSH antibody of the present invention may be the same as the sesame extract production reactive with the anti-HSP antibody and the anti-FGF antibody.
In order to produce root sesame having reactivity with the anti-MSH antibody, in addition to the light irradiation conditions described above, as in the case of producing an extract reactive with the anti-HSP antibody and anti-FGF antibody, Appropriate considerations are necessary for temperature management, water supply, ventilation, etc. The culture temperature of sesame is preferably selected under conditions that approximate the natural environment when seed sesame is sown in the field in early spring, and 25-30 ° C is appropriate. From the point that α-MSH is one of stress hormones, it is preferable to give a shock such as a low-temperature (or high-temperature) shock or some stress during sesame culture, but it is not particularly limited.
As a method of giving a shock by culturing at different temperatures, the culture is performed at 25 to 30 ° C., which is an appropriate temperature in normal sesame culture, and 5 to 20 ° C., preferably 5 from the lower limit of the appropriate temperature. Cultivation is performed at 5 to 20 ° C, preferably 15 to 20 ° C, which is a temperature lower by 10 ° C. The order of culturing at an appropriate temperature and culturing at a low temperature is not particularly limited, but first, cultivating at an appropriate temperature and rooting sesame, then culturing at a low temperature, and if necessary, culturing at an appropriate temperature. It is preferable to return. The culture time at a low temperature is not particularly limited, but is in the range of 1 to 10 hours, preferably 3 to 7 hours, more preferably 5 to 6 hours.
Conditions such as water and ventilation may be appropriately set depending on the seed sesame used. In general, it is sufficient that the water content is such that sesame seeds do not dry. When water is used, water is appropriately changed so that the water does not change in quality, but mist or water vapor can be used instead of water. The water used may be tap water, but if there is a problem with the quality of the tap water, it is better to use other appropriate water, and water containing mineral or organic components such as groundwater, mineral water, ocean Deep water or hot spring water may be used. Since the oxygen demand of the rooted sesame increases with time, it is desirable to supply oxygen corresponding to the time and the density of sesame.
The “extract” of rooting sesame is desirably a filtrate obtained from a homogenate of rooting sesame or a supernatant obtained by centrifugal precipitation thereof, but by other methods. Extraction may be performed using an aqueous solvent. In the case of centrifugal sedimentation, it is obtained by treating with a centrifuge, preferably a refrigeration high-speed centrifuge, with a rotational speed of 5,000 to 10,000 G and a duration of 10 to 30 minutes. Preferable separation conditions must be determined appropriately depending on the type of root sesame, growth status, homogenate status, type of centrifuge, etc., but the target separation conditions are from root sesame homogenate, shell fragments, nuclei, The cell wall debris is removed as a precipitate, and a Golgi apparatus containing a substance that reacts with an anti-α-MSH antibody, a secretory granule, or a supernatant that leaves a soluble fraction in the cytoplasm is obtained.
The root sesame extract used in the present invention contains a substance that reacts with an anti-α-MSH antibody. The inventor will refer to these substances herein as α-melanocyte stimulating hormone (α-MSH), but this term is also used in general for melanocyte stimulating hormone.
The substance that reacts with the anti-α-MSH antibody is preferably derived from rooting sesame seeds cultured under light irradiation, but is derived from other plants and animals, or from microorganisms such as yeast. Even if it is a thing, as long as the same effect can be anticipated, it can be used.
α-MSH often uses an acetate buffer when it is extracted from cells in a laboratory. As disclosed in the examples of the present invention, a particularly favorable effect is obtained for extraction with cold water. In addition to substances that react with anti-α-MSH antibodies, rooting sesame extract contains proteins useful for hair follicles such as heat shock proteins and fibroblast growth factors as described above. As the solvent, it is preferable not to use a solvent that solidifies the protein. In addition, since these useful proteins are susceptible to degradation and denaturation, measures to prevent them, for example, when using an organic solvent, reduce the concentration, protect with microcapsules or liposomes, or freeze. This can be stably provided as a cosmetic by drying or vacuum heating treatment.
By supplementing the scalp with an extract containing a substance that reacts with anti-α-MSH antibody that can be obtained from cultured sesame new roots, the activity of hair follicle melanocytes with reduced melanin synthesis ability is restored, Prevent and improve gray hair. That is, a substance capable of reacting with an anti-α-MSH antibody, which is one of the melanocyte-stimulating hormones of plant cells, can be used in humans to obtain a hair cosmetic material with improved gray hair prevention and excellent hair follicle melanocyte cell activation activity. The root sesame used preferably in the present invention has a new root length of 1 to 10 mm, preferably 3 to 5 mm, but is not particularly limited.
As described above, sesamin, which is widely known as a component of sesame, disappears on the 2nd to 3rd days from the rooting of sesame, and therefore this component cannot be expected from the rooting sesame in the present invention. Sesame oil is also consumed as an energy source in the process of rooting and disappears, so it is not included in the rooted sesame extract in the present invention.
When sesame roots and new roots reach 3 to 5 mm, the culture is stopped, and care is taken not to damage the new roots to make a new root homogenate. May be homogenized as it is. The substance that reacts with the anti-α-MSH antibody is contained in sesame new root and has a biochemically stable property. However, in the immunohistochemical study conducted by the present inventor, fibroblast growth factor Is also found in tissues other than new roots, it is better to homogenize the whole rooted sesame. When homogenizing rooted sesame, the homogenate is a substance other than α-MSH and has a mechanism of action different from that of α-MSH, but HSP25, HSP27, which is an active ingredient having an effect of improving white hair prevention, or FGF2 having a hair-growth effect. In order not to waste the effects of these proteins, it is preferable to take measures to prevent their degradation and denaturation. For that purpose, simply freeze the rooted sesame and lower the homogenization temperature to near freezing point.
Particularly preferred is a filtrate obtained from a homogenate of rooting sesame or a supernatant obtained by centrifugal precipitation thereof, but it may be extracted using an aqueous solvent by other methods. . This rooted sesame extract can be used as a raw material in hair cosmetics. When preserving the stock solution of rooting sesame extract, it is desirable to prevent decomposition and denaturation of the active ingredient by freezing. Usually, such rooted sesame extract contains 0.01 pg to 100 μg of anti-α-MSH antibody and / or a substance that reacts with anti-HSP25, anti-HSP27, anti-FGF2 or anti-FGF4 antibody per ml of the extract. Yes.
The present invention also provides a gray hair prevention / improving agent comprising a substance that reacts with an anti-α-MSH antibody derived from rooting sesame seeds cultured while being irradiated with light. This improvement in preventing white hair means that hair follicle melanocytes that have deteriorated in function are stimulated to recover their functions, thereby preventing the generation of white hair or those that have become white hair.
Furthermore, the present invention can provide a rooted sesame extract containing a useful protein such as HSP25, HSP27, FGF2 or the like as a hair cosmetic by adding a device for preventing protein denaturation in the process of producing the extract. HSP25 and / or HSP27 have the effect of preventing and improving white hair as in the present invention by preventing apoptosis of hair follicle melanocytes, and FGF2 has the effect of hair growth by activating the hair papilla. is there.
When rooting sesame extract is blended into hair cosmetics, the amount of rooting, the rooting state, the type and amount of light applied during cultivation, the extraction method of the extract, the extraction efficiency and the active ingredient stabilization method, etc. Depending on the subject of use and the cosmetic preparation, it can be set as appropriate depending on various circumstances. For example, when a root sesame extract having approximately the same mass as the wet mass of root sesame is produced, the extract can be blended into the hair cosmetic at a ratio of 10 to 50% by mass. In order to maintain the physiological activity of the active ingredient, the rooted sesame extract may be stored frozen, freeze-dried, protected with microcapsules or liposomes, or vacuum heated for sterilization purposes.
The hair cosmetics of the present invention include fats and oils such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, A sequestering agent, a polymer such as a water-soluble polymer, a thickener, a powder component, an ultraviolet absorber, an ultraviolet blocking agent, a fragrance, a pH adjuster, and the like can be contained. In addition, other medicinal components such as vitamins, skin activators, blood circulation promoters, active oxygen scavengers, anti-inflammatory agents, and bactericides, and physiologically active components can be contained.
The present invention further provides a pharmaceutical composition characterized by containing, as an essential component, a rooted sesame extract cultured while being irradiated with light.
The hair cosmetic composition and pharmaceutical composition of the present invention can be applied in the form of a semi-solid agent such as an aqueous solution, an oil agent, a milky lotion, a liquid solution such as a suspension, or a gel or cream. It can be prepared in these forms by a conventionally known method to form various dosage forms such as lotions, emulsions, gels, creams and aerosols. These can be applied to the scalp by spraying or spraying. Of these dosage forms, lotions, emulsions, creams, aerosols and the like are particularly suitable. Usually, these dosage forms can be produced according to the formulation method used in cosmetics and pharmaceutical compositions.
Furthermore, since sesame is originally a food, rooted sesame can be used as a food or supplement as well as bean sprouts made from soybeans. At that time, it is preferable to prevent the physiological activity of the active ingredient from being lowered by performing a treatment such as freezing or freeze drying. From a hygienic standpoint, vacuum heating may be performed for sterilization purposes.
As food, it is eaten as a health food or food material useful for prevention or treatment of gray hair, as it is, or by adding various nutritional components or adding it to foods and drinks. After adding suitable starch, vegetable oil and other auxiliaries, it can be used for edible by using conventional means to form it into an edible form, such as granules, granules, tablets, capsules, pastes, etc. In addition, it may be used by adding to various foods such as bread and fermented dairy products, or by adding to beverages such as soft drinks. It can also be used in the form of an extract.
EXAMPLES Hereinafter, although an experimental example, a reference example, and an Example are given and this invention is demonstrated in detail, this invention is not limited to these Examples.
Reference example 1
How to determine the quality of new roots of rooted sesame
The determination of the quality of sesame new root tissue or cells not only determines the quality of the active ingredients of rooted sesame, but if the new root is denatured, it may be harmful due to the formation of skin irritants. It is important because it can be. The most common damage to sesame new roots is degeneration or necrosis of new root cells when viewed pathologically. The cause of degeneration or necrosis of sesame new roots is due to defects in water quality used for sesame rooting, inadequate exposure to light, excessive or insufficient oxygen, inadequate temperature, contamination due to growth of microorganisms such as mold, and other factors . Degeneration and necrosis of new root cells are uncertain with the naked eye, and it is certain that a tissue specimen is prepared and determined microscopically. Microscopic findings include cytoplasmic edematous degeneration and fatty degeneration (appearance of fine lipid droplets) as degeneration, and liquefied necrosis as necrosis. In order to check whether degeneration or necrosis has occurred, a part of the rooted sesame seed was collected as a sample, tissue sections were prepared through formalin fixation, ethanol dehydration, and paraffin embedding, and hematoxylin and eosin double Dyeing and observation with a microscope. If the production status of the active ingredient is judged, immunohistochemical judgment on the tissue section is effective, but the procedure is complicated and the necessary cost is high. Therefore, as a convenience method, firstly, the state of sesame new root was visually observed to confirm a faint pale yellow coloration by melanin, and then judged by hematoxylin-eosin double-stained specimen, One method is to determine whether the tissue cells are growing normally and to infer that the active ingredient would have been produced.
Experimental example 1
Immunohistochemical studies on HSP25, HSP27, FGF2 and FGF4 in human scalp hair follicles:
A surgical material for human scalp was fixed with 10% formalin solution, applied to an automatic dehydration embedding apparatus, and then embedded in paraffin to prepare a paraffin block. Using a microtome, a 5 micron thick scalp tissue section was prepared from this paraffin block, which was attached to a slide glass and dried. The tissue sections were deparaffinized with xylol, anti-HSP25 antibody (Funakoshi SPA-801), anti-HSP27 antibody (Cosmo Bio SC-1049), anti-FGF-2 antibody (Cosmo Bio SC-79G) and anti-FGF-4 antibody ( Each tissue section was subjected to immunohistochemical staining (hereinafter referred to as “immunostaining”) by ABC method (Avidin-Biotin complex method), which is a general immunostaining method, using Cosmo Bio SC-1361). went. When the hair follicle tissue was examined microscopically with respect to the stained scalp tissue specimen, the cytoplasm of the cells constituting the hair bulb was stained brown, and it was found that HSP25 and HSP27 were positive (FIG. 1). In the stained specimens for HSP25 and HSP27, no difference could be found between them. Although it is already known that the cytoplasm of outer root sheath cells is HSP27 positive, it is a new finding that the constituent cells of the hair bulb are HSP positive. No immunostaining of hair follicle tissue with HSP25 has ever been reported. Whether the melanocytes in the hair bulb are positive for HSP25 or HSP27 cannot be determined because the cytoplasm of the melanocytes is black, and the presence or absence of brown color by immunostaining cannot be identified. In the immunostaining for FGF-2, cells constituting the hair bulb and root sheath showed positive findings, and cells of the hair papilla also showed positive results (FIG. 2). Immunostaining for FGF-4 showed positive findings similar to FGF-2, but cells on the hair papilla showed negative findings.
Experimental example 2
Examination by immunostaining for HSP25 and HSP27 in human placenta:
Since HSP25 and HSP27 are female hormone-dependent heat shock proteins, it was decided to confirm the localization of HSP25 and HSP27 in a placenta that safely raises a fetus with the support of female hormones. A fresh human placenta was fixed with 10% formalin solution, a placental tissue fragment containing a decidua was cut out, a placenta tissue section was prepared in the same manner as in Example 1, and immunostaining for HSP25 and HSP27 was performed. . When examined microscopically, the cytoplasm of the decidual cell (Fig. 3) and the cytoplasm of the placental choriotrophic cell (trophoblast) (Fig. 4) were stained brown and exhibited HSP25- and HSP27-positive findings. .

How to make root sesame seeds with temperature shock:
Black and white sesame seeds were purchased from a seed store. Seed sesame seeds were spread on a stainless steel sieve, washed gently with water, placed in a flat plastic container, and mineral water was added until the sesame seeds were sufficiently immersed. When this is cultured at 25 ° C. for about 40 hours and at 30 ° C. for about 30 hours, the majority of new sesame roots reach 1 to 2 mm. At this time, the cells were cultured at 15 ° C. for 5 to 6 hours as a cold shock. When the length of the new root reached 2 to 3 mm, the culture was stopped, the water was drained lightly so as not to damage the new root of the rooted sesame, and frozen at -20 ° C.

Examination by immunostaining for HSP25, HSP27, FGF-2, FGF-4 and MIB1 in tissue cells of rooting sesame:
About 1 gram of rooting sesame prepared by applying a low temperature shock in Example 1 was collected as a sample, fixed with a phosphate-buffered 10% formalin solution containing salt, and rooted in the same manner as in Example 1. Sesame tissue sections were prepared and immunostained for HSP25, HSP27, FGF and FGF-4. Furthermore, in order to recognize the proliferating cell group undergoing nuclear division in the rooted sesame tissue, immunostaining was also performed using an anti-Ki-67 antibody (Cosmo Bio PRO229) against MIB1, which is a nuclear protein in the mitosis phase. .
When a stained tissue sample of rooted sesame is observed with a microscope, the cytoplasm of the surface cells at the tip of sesame new root is stained brown in both of the stained samples for HSP25 and HSP27, and positive findings are observed for HSP25 and HSP27. (FIG. 5) In the immunostaining for nuclear protein antigen MIB1 in the mitotic phase, not only the surface cells at the tip of the new root, but also the whole tip and the nucleus of the cells in the medullary region showed MIB1 positivity. (FIG. 6). From this staining result, it is considered that the cells on the outermost layer produced HSP25 and HSP27 in order to protect the proliferating cells of the new root tip and the medulla from low temperature stress among the cells proliferating while undergoing nuclear division. It was.
On the other hand, in the stained specimen for FGF-2, the positive cells are distributed in the cytoplasm and nucleus of the cells of the new root surface and medulla (FIG. 7), and further branch and correspond to dendritic or primitive veins inside the cotyledons. It was distributed in the structure (FIG. 8). FGF-2 is a basic fibroblast growth factor. In animal tissues, it has been known to proliferate to various mesodermal cells such as an angiogenic factor, but its expression in plants has been reported. I do not see. In a rooted sesame tissue, as seen from a microscopic image in which FGF-2 positive cells are stained in accordance with a structure corresponding to a primitive vein, these cells are likely to be involved in tissue nutrient transport. In animal (rat) hair follicles, FGF-2 has been reported to stimulate and proliferate cultured dermal papilla cells (Matsuzaki T, Inamatsu M, Yoshizato K: Hair induction by dermal peels citrated cultivated cells) medium of keratinocytes.In, van Nest DJJ & Randall VA (Eds), Hair Research for the Next Millennium (Processings of the First Tritontinent. Even in the FGF-2 staining of the human scalp hair follicle in Example 1, FGF-2 may be involved in the hair follicle papilla of the human scalp because the hair papilla cells showed a positive finding. On the other hand, immunostaining for FGF-4, unlike FGF-2, did not find a strong positive part in new root sesame, and a weak positive finding in medullary cells. In the literature, FGF-4 knockout mice are unable to develop embryonic cell clusters and cause fetal death (Feldman B et al. Science 267: 246-9, 1995), and are therefore involved in the development of juvenile cells. It is said to be a growth factor. The reason for the low FGF-4 positivity in the root sesame seeds this time is considered to be that the development of root sesame seeds has progressed and the primitive juvenile period has passed.

Production of rooted sesame extract The process of producing an extract from the frozen rooted sesame produced in Example 1 will be described. First, the wet mass of rooted sesame was weighed, and the same amount of cold mineral water was added, and the mixture was homogenized with a high-speed homogenizer (Hiscotron) at a maximum speed of 15,000 rpm under ice cooling. The homogenate was spun down at 4O <0> C and 10,000g for 30 minutes in a frozen ultracentrifuge. The supernatant became an almost colorless and transparent aqueous solution even in the case of black sesame. This supernatant was used as a root sesame extract, dispensed into small containers and stored frozen at -20 ° C.

Experiments to see the effect of scalp lotion containing rooting sesame extract on improving gray hair:
The root sesame extract stock solution prepared in Example 3 was diluted 4 times with mineral water, and this was used as the scalp lotion of the root sesame extract. A number of 2 ml each of the same lotion dispensed into a small container and stored frozen was prepared, and when applying to the scalp, this was thawed one by one and used. A subject with white hair mixed hair was applied to the scalp once a day after shampooing. After the germination sesame solution application was continued for 2 months, the hair was removed from a certain small portion, and changes in the hair were first visually observed. As a result of observation, it was recognized that the hair was white and the root was black. Those with white hair are considered as resting hair on the hair cycle because the roots of the hair are swollen in a club shape. It was recognized that the hair with the hair root not swollen was full-haired and the hair near the root was blackened. In order to examine whether the blackened part is due to melanin, this hair was fixed with a phosphate buffered 10% formalin solution with salt added, pasted on a slide glass with gelatin solution, and melanin stained with Fontana silver stain. And observed with a microscope. As a result, the blackened portion at the base of the hair was darkened so strongly that the fur quality was completely indistinguishable from the pulp, and it was proved that this blackened portion contained melanin (FIG. 9). ). At the tip of the gray hair that did not turn black, the fur was gray and only the medulla was intermittently black (FIG. 10). The length of the blackened hair root determined to be melanin positive is 1 to 1.2 cm, and the average length of hair growth is 0.3 to 0.4 mm per day. It was presumed that the application of lotion resulted in remelanization of the root of this gray hair. The reason why black hair was not found in the resting hair was because hair follicle melanocytes disappeared in the resting hair, and the melanin reproduction did not occur even by applying rooting sesame extract / lotion. I guessed it. In addition, the reason why some hair blackened and not blackened was found in the growing hair. The recovery of inactivated hair follicle melanocytes can be fast or slow. It was speculated that blackening of gray hair was observed. Even in the case of hair follicle melanocytes that are slow to recover, there is a possibility of recovery by continuous application of rooting sesame lotion for 2 months or more. By applying the rooting sesame lotion of the present invention every day, signs of white hair improvement appear 2 months at the earliest time, but 3-4 months or more at the later time. Since the improvement of gray hair does not occur with resting hair, it is envisioned that older people with more resting hair follicles will need more time to recover than younger ones.

Hair growth effect assessment by hair loss test:
In order to judge the hair-growth effect by application of root sesame extract, a hair loss test was performed. One male in his 30s and two males in his 40s, every day after a night shampoo and twice in the morning, 1 scalp lotion containing the same rooting sesame extract used in Example 6 -The experiment of applying 1.5 ml to the scalp was continued for 2 months. The number of hair loss was calculated before and after the start of the experiment and once each time. As in the mold, the method was laid on a hair-washing sink using a stocking fabric as a filter, and after shampooing, the total number of hair loss was counted. In the measurement before the start of the experiment, the total number of hair loss of the three subjects was 120, 98, and 92, respectively, but after the experiment was 66, 70, and 61, respectively, and the reduction of hair loss was obvious. From this experiment, it was proved that the scalp lotion containing the rooted sesame extract prevents hair cycle resting period and has a hair growth effect.

Production method of rooting sesame (culture while shining light and culturing while blocking light)
Seed sesame (hereinafter, sesame) was purchased from a seed specialty store. Sesame seeds are cultivated while exposed to light, and cultivated while blocking light. Place them on separate stainless steel sieves, wash them roughly with water, put the sieves in a plastic container, and add mineral water or deep ocean water. Was added until the sesame was sufficiently immersed, and the sesame was allowed to absorb water for about 15 hours. Then, sesame was cultured at 25 ° C. while supplying water to the small air pump. In the case of sesame seeds that were cultivated under light, they were exposed to sunlight when new roots started to break through the shells.
In the summer, direct sunlight is not exposed to sesame for a long time in the summer so that it is placed on a bright window. In the case of sesame culture without light, which was a control experiment, a cover made of thick black paper was covered with a sesame culture vessel to block the light. In any case, when the length of new roots reached 2 to 3 mm after culturing for about 40 hours, the cells were cultured at 15 ° C. for 5 to 6 hours while supplying air to give a low temperature shock. Thereafter, the culture conditions were again returned to 25 ° C., and when the length of the new root reached 3 to 5 mm, the culture was stopped and washed with water with care not to damage the new root. For making the rooted sesame extract, the rooted sesame was frozen at -20 ° C. The portion for making the microscopic tissue specimen was fixed in formalin in a 10% formalin solution supplemented with saline. The new roots of sesame seeds cultured with light were pale yellow, but the new roots of sesame seeds cultured under the light were white. (See Figure 11)
The root sesame tissue structure is shown in FIG.

Staining of Root Sesame Melanin About 1 gram of root sesame prepared in Example 6 and fixed with formalin solution is placed in a sample cassette, dehydrated and paraffin infiltrated with an automatic tissue dehydration embedding device, and then embedded in paraffin Thus, a paraffin block was produced. Using a microtome, a tissue section of rooted sesame having a thickness of 5 microns was prepared from this paraffin block, and was attached to a slide glass and dried. The slices were processed by deparaffinization, ethanol series, and tap water by a conventional method, and stained with melanin by Masson's Fontana silver method. When the stained specimen was observed with a microscope, it was confirmed that the cytoplasm of the epidermis cells of sesame new root cultured under light was stained strongly black and melanin was positive (see FIG. 13).
On the other hand, in the specimen of rooted sesame seeds cultured in the dark, the melanin-positive findings of epidermis cells of sesame new roots were extremely weak (see FIG. 15). In sesame seeds, melanosomes found in humans are not found, so it is likely that epidermal cells produce melanin. When comparing the melanin positivity between black sesame and white sesame, white sesame was stronger.

Examination of rooted sesame by α-MSH immunostaining The rooted sesame prepared in Example 6 and fixed with formalin solution was treated in the same manner as in Example 7 to prepare a tissue section, and anti-α-MSH antibody (Cosmo Bio AB946) was used for immunostaining by the usual ABC method. When the stained tissue specimen is observed under a microscope, in the sesame seeds cultured with light, the cytoplasm of the inner sheath cell pericule (or the innermost endothelial cell endodermis) of the central column of the new root is stained brown and α- MSH was positive (see FIG. 14). In sesame seeds cultured in the dark, the number of α-MSH positive cells of new roots was small, and the degree of positivity of individual cells was weak.
From the results of melanin staining in Example 7 and α-MSH immunostaining in Example 3, α-MSH secreted from α-MSH-secreting cells inside the new root of rooting sesame cultured under light irradiation, It was assumed that melanin was produced by acting on epidermis cells of sesame new root. Regarding the effect of low temperature shock during culture on α-MSH production, sesame cultured with low temperature shock tended to have higher α-MSH positivity than sesame cultured without low temperature shock. The low temperature shock may have promoted the production of a substance that reacts with the anti-α-MSH antibody.

Examination of rooted sesame by electron microscope In order to confirm melanin staining positive cells and α-MSH positive cells of new roots of rooted sesame proven in Example 7 and Example 8 using a transmission electron microscope, direct sunlight A fresh tissue piece of sesame radish rooted on the surface was fixed with glutaraldehyde / osmium by a conventional transmission electron microscope sample preparation method, dehydrated with an ascending ethanol series, and ethanol was added with propylene oxide. Replaced, infiltrated with an epoxy resin preparation for electron microscope (hereinafter referred to as Epon), embedded in capsules, made Epon block by thermal polymerization of Epon, and made root sesame from Epon block using ultratome and diamond knife Prepare an ultrathin tissue section of the root, place it on a copper grid, double-stain uranium / lead acetate, and observe with a Hitachi electron microscope Was Tsu.
When the epidermis cells of the colored portion near the root of sesame new root were observed, many small granules with a high electron density of 0.5 to 0.05 microns in diameter that seemed to be melanin in the cytoplasm, and the nuclear membrane Along the high electron density material was observed (see FIG. 16a). When the sesame new root part in which α-MSH positive cells were observed with an optical microscope in Example 3 was observed with an electron microscope, cells containing abundant Golgi apparatus and secretory granules were found (see FIG. 16 b).
In addition, in order to confirm α-MSH positive cells using immunoelectron microscope technology, fresh tissue pieces of sesame new roots rooted in direct sunlight were fixed with phosphate buffered saline 0.4% in paraform. After fixing with a solution, a tissue section of rooted sesame was prepared by the same method as in Example 1, and immunostaining for α-MSH was performed. After confirming α-MSH positive cells with an optical microscope, the tissue specimen was dehydrated with ethanol and propylene oxide, infused with Epon, and a prepolymerized Epon block was adhered onto the α-MSH positive cells. The thermal polymerization of was completed. The Epon block containing α-MSH positive cells was peeled off from the slide glass, and using this Epon block, an ultrathin section was prepared by the same method as described above, and observation with a Hitachi electron microscope was performed without electron staining. On the electron microscope, many small granules having a high electron density of 120 to 150 nm in diameter were found in the cytoplasm of α-MSH positive cells. Since these small granules were α-MSH positive, they were determined to be α-MSH granules (FIG. 16c). reference).

Production of rooted sesame extract The process of producing an extract from the rooted sesame produced in Example 6 will be described. First, the wet mass of rooting sesame was weighed, the same amount of mineral water (or deep sea water) was added, and homogenized at a maximum speed of 15,000 rpm with a high-speed homogenizer (Hiscotron). The homogenate was centrifuged with an ultracentrifuge at 4 ° C. and 8000 G for 20 minutes. The supernatant became an almost colorless and transparent aqueous solution even in the case of black sesame. This supernatant was used as a root sesame extract, dispensed into small containers and stored frozen at -20 ° C.

White hair improvement effect of lotion containing rooting sesame extract The rooting sesame extract undiluted solution prepared in Example 6 and Example 9 was diluted twice with mineral water to form a rooting sesame extract lotion, which was reduced by 2 ml. Dispensed into containers and stored frozen. The effect of the lotion on improving gray hair was examined using gray hair and eyebrows. Unlike the hair that grows two to three from one place, the eyebrows grow independently one by one, are short in length, and are advantageous in that the color change at the base can be observed in detail. The same lotion that had been dispensed into small containers and stored frozen was thawed and used one by one. A male subject with white hair mixed hair was applied to the scalp once a day after shampooing. In the eyebrows, 1-2 drops of the same lotion were rubbed.
After applying rooting sesame solution for 2 months, in the case of hair, remove it from a certain small part, and in the case of eyebrows, remove only white hair or hair that has undergone a color change and visually observe it. did. Some of the hair was white hair, grayish white, and the roots were black to various degrees. Since the eyebrows were thicker than the hair, the changes could be observed more clearly. FIG. 17 shows an example of eyebrows with the white hair removed and eyebrows with a black base. The white eyebrows were judged to be resting on the hair cycle from the point that the root ends were swollen. In the hair cycle resting phase, the melanocyte cycle also enters the resting phase, and it is presumed that melanin reproduction did not occur because the hair follicle melanocytes disappeared at the hair root ends.
On the other hand, those whose hair roots were darker were judged to be in the growing stage on the hair cycle from the point that the hair root ends swelled up like a cup upside down. Although this white hair had stopped melanin production, it seems that melanin production was restarted by root sesame lotion application. The length of the hair root part that has turned black halfway is 1 to 1.2 cm, and the average growth of hair is 0.3 to 0.4 mm per day, so the application period of rooting sesame lotion for 2 months It is presumed that resumption of melanin production by hair follicle melanocytes occurred at any of these times. On the other hand, some hairs, which are judged to be growing from the shape of the ends of hair, were not blackened at all, and some were blackened to various degrees.
The reason for the variety of changes is that recovery of inactivated hair follicle melanocytes can be fast or slow, with rapid recovery causing melanin regeneration within the two-month use period of the lotion. However, it was speculated that the slow recovery was insufficient. Even hair follicles with slow recovery of hair follicle melanocytes can be recovered by applying the same lotion for 2 months or more. Therefore, it seems that the daily application of rooting sesame lotion of the present invention shows signs of improvement in gray hair within 2 months at the early stage, but 3-4 months or longer at the later stage. Re-melanization cannot occur in resting gray hair, but re-melanization can occur in nascent hair that grows by generational change, while resting gray hair falls off. Therefore, it seems that in elderly people with a high content of resting hair follicles, recovery of gray hair will take longer than in younger people.

Production example using a fluorescent lamp as a light beam to irradiate sesame Using a plant incubator, using a fluorescent light beam as a light beam to be applied to sesame, changing the intensity, the fluorescent light beam is melanin and anti-α- An experiment was conducted as to whether or not production of a substance that reacts with the MSH antibody was caused.
Preparation: A plant incubator (CFH-305 manufactured by Tommy Industries) (hereinafter referred to as an incubator) using a fluorescent lamp (Toshiba fluorescent lamp mellow 5w, light source color EX-W (three-wavelength white)) as an irradiation light source was purchased. In this incubator, the temperature, illuminance, and humidity in the culture apparatus can be freely set in advance by dividing time. Therefore, in this experiment, a 6-step culture condition was set, imagining the natural environment of the season when sesame seeds were sown in the field. As the sesame culture container, the same stainless steel sieve and water container as in Example 6 were used. Tap water was used as the culture water. In the incubator, two sesame culture vessels were placed on each shelf, up to two. The maximum dry mass when the sesame seeds were not overlapped in one sieve was 18 g.
Culture conditions: Sesame water absorption started at 9:30 am. The incubator temperature is 25 ° C., which is the same as room temperature, and the illuminance is 0.
Step 1. At 10:00 am for 30 minutes, the inside of the incubator was adjusted to a temperature of 25 ° C. and an illuminance of 2000 lux. Humidity was 60% throughout all steps.
Step 2.2 At noon over an hour, the inside of the incubator was brought to a temperature of 30 ° C. and an illuminance of 2000 lux.
Step 3.3 Over 3 hours at 3:00 pm, the inside of the incubator was brought to a temperature of 30 ° C. and an illuminance of 4000 lux.
Step 4.9 The temperature in the incubator was set to 20 ° C. and the illumination intensity was set to 0 midnight at midnight.
Step 5. Thereafter, the inside of the incubator was maintained at a temperature of 20 ° C. and an illuminance of 0 lux until 6 am for 6 hours.
Step 6.3 Over 9 and a half hours, the temperature inside the incubator was 25 ° C. and the illumination intensity was 2000 lux at 9:30 am.
Thereafter, Steps 1 to 6 were repeated.
Sesame macroscopic findings:
In the morning of the second day of culture (24 hours after the start of culture), roots were observed in some sesame seeds.
In the morning of the third day of culture (48 hours after the start of culture), more than 80% of sesame roots were found, and the length of new roots reached a maximum of 5 mm.
Immunostaining of rooted sesame and its evaluation:
A portion of the rooted sesame seed was fixed in a phosphate buffered 10% formalin fixative as a sample, and stained with Masson's Fontana silver method and FGF2, HSP25, HSP27 and α in the same manner as in Examples 2 and 3. -Immunostaining for MSH was performed. In the microscopic evaluation for each stained specimen, the new root of sesame cultured while applying fluorescent light in this plant incubator was compared with the new root of sesame cultured under natural light in the room according to Example 1. In immunostaining specimens for FGF2, HSP25, HSP27 and α-MSH, both showed almost the same staining results. However, in the melanin-stained specimen by Masson's Fontana silver method, the new root obtained this time showed a stronger positive finding than the case of Example 1. The reason was interpreted that the brightness of the maximum illuminance of 4000 lux determined by assuming the natural world was stronger than the illuminance in the room in Example 6.

常法に従って、各成分を混合し、実施例１４では糖度１６、酸度０．６３、ｐＨ３．４、実施例１５では糖度６、酸度０．６３、ｐＨ３．２６のドリンク剤を得た。これを９５℃で３０秒間加熱し、ドリンク剤用容器に充填し、所望の発根ゴマ抽出物配合ドリンク剤を得た。このドリンク剤は１日３０〜３６０ミリリットル服用することにより、すぐれた白髪改善と育毛促進効果が発揮される。Changes in substances reactive with anti-MSH antibody due to differences in rooting sesame culture conditions and extraction conditions In the same manner as in Example 6, rooting sesame was cultured using a plant incubator. Two days after the start of the culture, sesame was collected and used for extraction. The culture conditions were as follows.
Condition 1 Start of rooting on June 16, 2003 Black sesame No aeration Tap water Humidity 60%
Condition 2 Start rooting on June 23, 2003 White sesame No aeration Tap water Humidity 60%
Condition 3 Start of rooting on September 8, 2003 Black sesame aeration available Deep sea water Humidity 35%
Condition 4 Start rooting on September 29, 2003 Black sesame aeration available Deep sea water Humidity 60%
Extraction method Sesame rooted was immediately freeze-dried (freeze dryer FRD-80, Iwaki Glass Co., Ltd., Chiba) and stored in a freezer until the start of the test. The dried root sesame was pulverized by a mill and used for extraction. About 5 g of pulverized sesame seeds was used a high-speed solvent extraction device ASE-200 (Nippon Daionex Co., Ltd., Osaka). Extraction was performed at 1500 psi, TEMPERATURE 0 ° C. (100 ° C. in the case of hot water). Extraction solvent (cold water, hot water, 10% ethanol, 50% ethanol, 100% ethanol, 50% 1,3 butylene glycol, 1% acetic acid solution, 3% acetic acid solution, 5% acetic acid solution) is distilled water production apparatus GSR- Distilled water prepared in 200 (Advantech Toyo Co., Ltd., Tokyo) was used, and a commercially available (special grade) organic solvent was used. The extracted solution was concentrated using a rotary evaporator NE (Tokyo Rika Kikai Co., Ltd., Tokyo) and freeze-dried. The extract was converted from dry mass, and the recovery rate was 4 to 7%.
Quantification of the substance that reacts with the anti-α-MSH antibody was performed using α-MSH EIA kit (PHOENIX PHARMACEUTICALS, INC.). Each extract was dissolved in a solvent attached to the kit at a concentration of 10 mg / ml, the insoluble part was removed by centrifugation, and the supernatant was used for measurement.
Content of substance that reacts with anti-MSH antibody Substance that reacts with anti-MSH antibody in rooted sesame extracted with cold water, hot water, 10% ethanol, 50% ethanol, 100% ethanol, 50% 1,3-butylene glycol It measured by EIA method of (alpha) -MSH. The sesame of the condition 1 was used for the measurement. As a result, the amount of α-MSH in the cold water was the largest (FIG. 18).
In addition, since α-MSH was dissolved in the acetic acid solution, the amount of the substance that reacted with the anti-α-MSH antibody in the rooting sesame extracted with cold water and the acetic acid solution was similarly compared. Sesame of condition 3 was used for the measurement. As a result, it was found that extraction with cold water was easier than with acetic acid solution (FIG. 19).
The amount of the substance that reacts with the anti-α-MSH antibody in sesame rooted under the four conditions shown in the above method was measured by EIA. It is considered that the amount of α-MSH is higher in white sesame than in comparison between condition 1 and condition 2. In addition, the amount of α-MSH was the highest in condition 3 using aeration and deep ocean water (FIG. 20).
Using the root sesame extract obtained in Example 6, the drink of the present invention was prepared according to the formulation shown in the following “Table 1”.

In accordance with a conventional method, each component was mixed, and in Example 14, a drink having a sugar content of 16, a acidity of 0.63, pH 3.4, and in Example 15, a sugar content of 6, a acidity of 0.63, and a pH of 3.26 was obtained. This was heated at 95 ° C. for 30 seconds and filled into a drink container to obtain a desired rooted sesame extract-containing drink. By taking 30 to 360 milliliters of this drink per day, excellent white hair improvement and hair growth promoting effects are exhibited.

  According to the present invention, an agent for improving and preventing gray hair, which prevents or improves whitening of hair by restoring the activity of hair follicle melanocytes which have decreased melanin synthesis ability, stopped or cell death tendency, and gray hair A cosmetic for hair and a hair growth-promoting agent useful for prevention and improvement of hair are provided. In addition, it can be used for medicines and foods and drinks to contribute to the prevention and treatment of gray hair.

Claims (10)

It is reactive to any one or more of the group consisting of anti-heat shock protein (HSP) antibody, anti-fibrocyte growth factor (FGF) antibody and anti-melanocyte stimulating hormone (MSH) antibody. Rooted sesame extract. The rooted sesame extract according to claim 1, wherein the anti-heat shock protein (HSP) is HSP25 and / or HSP27. The root sesame extract according to claim 1 or 2, wherein the anti-fibrocyte growth factor (FGF) is FGF2 and / or FGF4. The root sesame extract according to claim 1, 2 or 3, wherein the anti-melanocyte stimulating hormone (MSH) is α-MSH. A root sesame extract characterized by having reactivity with one or more antibodies selected from the group consisting of HSP25 and / or HSP27 antibody, FGF2 and / or FGF4 antibody, and α-MSH antibody. A hair cosmetic comprising the rooted sesame extract according to any one of claims 1 to 5. An agent for preventing and / or improving white hair comprising the rooted sesame extract according to any one of claims 1 to 5. A pharmaceutical composition comprising the rooted sesame extract according to any one of claims 1 to 5. A hair growth promoter comprising the rooted sesame extract according to any one of claims 1 to 5. A food or drink comprising the rooted sesame extract according to any one of claims 1 to 5.

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