KR20230000935A - Composition for the prevention or treatment ofneurodegenerative disease or cognitive dysfunctioncomprising bamboo inner bark extract - Google Patents

Abstract

The present invention relates to a composition for preventing or treating neurodegenerative disease or cognitive dysfunction containing a Bambusae Caulis in Taeniam extract as an active ingredient. According to the present invention, the Bambusae Caulis in Taeniam extract, especially an ethanol extract of Bambusae Caulis in Taeniam extract contains high contents of p-coumaric acid and tricin, and was proved that an inflammatory response induced in microglial cells stimulated with LPS was effectively suppressed. In addition, since it was proved that the Bambusae Caulis in Taeniam extract exhibits a memory enhancing effect in a Y-maze test and a passive avoidance test using mice with memory impairment induced by scopolamine, and exhibits acetylcholinesterase inhibitory activity, antioxidant activity, lipid peroxide accumulation inhibitory activity, and activity to suppress the expression of inflammatory biomarkers, the Bambusae Caulis in Taeniam extract can be usefully used in pharmaceutical compositions or health functional foods for preventing or treating memory and cognitive dysfunction.

Description

Composition for the prevention or treatment of neurodegenerative diseases and cognitive dysfunction containing the extract as an active ingredient

The present invention relates to a composition for the prevention or treatment of neurodegenerative diseases, or a composition for the prevention or treatment of cognitive dysfunction, containing an extract as an active ingredient.

Microglia, phagocytes of the brain, can induce chronic neuroinflammation by producing superoxide and neurotoxin, and activators and neurotoxic substances produced by microglia are Parkinson's disease It is directly related to the pathological progression of diseases caused by neuroinflammation, such as

In particular, microglia activation is responsible for the pathogenesis of multiple neurodegenerative disorders such as Parkinson's disease (PD), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS). connected to the mechanism. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-12 (IL-12), interleukin-6 (IL-6) -6), prostaglandin E2 (PGE ₂ ), and cyclooxygenase-2 (COX-2). Other toxins have been established as neurotoxins such as Lipopolysaccharide, 6-OHDA and hydrogen peroxide (H ₂ O ₂ ). These toxins have been reported to stimulate microglia to activate inflammatory pathways such as NF-κB, p38, MAPK, JNK, ERK and IRF3 (Kim, DC, et al. , Molecules , 22(12):2130, 2017; He, X., et al. , Toxicology letters , 283: 58-68, 2018; Lim, H.-W., et al. , Food and chemical toxicology , 72: 265-272, 2014; Cho, D .-Y., et al. , Molecules , 21(12):1718, 2016). Therefore, modulating microglia activation can serve as an important therapeutic strategy to attenuate neuroinflammation.

Juk-yeo (竹茹, Bambusae Caulis in Taeniam) is the inner bark of bamboo. As for the efficacy of killeo, Donguibogam is known to be effective in symptoms such as vomiting, antipyretic, insomnia, and expectoration. Efficacy (Republic of Korea Patent Registration No. 10-1532307) and research into cosmetic ingredients (Republic of Korea Patent Registration No. 10-2082580) are being made.

Therefore, in the present invention, as a result of diligent efforts to develop an effective neuroinflammation inhibitor, the kill extract, especially the kill kill ethanol extract, contains a high content of p-coumaric acid and tricin, and is stimulated with LPS. It was confirmed that the inflammatory response induced in microglia was effectively suppressed.

In addition, the extract showed memory enhancing effects in the Y-maze test and passive avoidance test using mice whose memory impairment was induced by scopolamine, and acetylcholinesterase inhibitory activity, Antioxidant activity, lipid peroxide accumulation inhibitory activity, and expression inhibitory activity of inflammatory biomarkers were confirmed.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating neuroinflammatory diseases comprising an extract as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving neuroinflammatory diseases comprising an extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating memory and cognitive dysfunction comprising an extract as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving memory and cognitive function disorders comprising an extract as an active ingredient.

In order to achieve the above purpose,

The present invention provides a pharmaceutical composition for the prevention or treatment of neuroinflammatory diseases comprising an extract as an active ingredient.

In addition, the present invention provides a health functional food composition for the prevention or improvement of neuroinflammatory diseases comprising the kill extract as an active ingredient.

According to a preferred embodiment of the present invention, the extract is purified water (or water), lower alcohol having 1 to 4 carbon atoms, glycerin, ethyl acetate, acetone, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, It can be extracted with one or more solvents selected from the group consisting of butylene glycol, hexane, and mixed solvents thereof, and preferably with water or ethanol (alcohol).

According to another preferred embodiment of the present invention, the killing may be the inner skin of the royal family or the inner skin of the bamboo shoot.

According to another preferred embodiment of the present invention, the kill extract may contain coumaric acid (p-coumaric acid) or tricine (tricin) component.

According to another preferred embodiment of the present invention, the kill extract

i) inhibition of production of inflammatory mediators including NO, prostaglandin E ₂ , iNOS and COX-2;

ii) inhibition of production of inflammatory cytokines including IL-1β, IL-6 and TNF-α; and

iii) inhibition of IκB-α phosphorylation and degradation; can regulate neuroinflammation.

According to another preferred embodiment of the present invention, the neuroinflammatory disease is multiple sclerosis, neuroblastoma, ischemic stroke, Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, Huntington's disease, Creutzfeldt-Jakob disease, post-traumatic stress disorder, depression, mental schizophrenia or amyotrophic lateral sclerosis.

to achieve other purposes,

The present invention provides a pharmaceutical composition for the prevention or treatment of memory and cognitive dysfunction comprising an extract as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving memory and cognitive function disorders, including the extract as an active ingredient.

According to a preferred embodiment of the present invention, the extract can be extracted with one or more solvents selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, ethyl acetate, methylene chloride and mixed solvents thereof, , preferably by water or ethanol.

According to another preferred embodiment of the present invention, the killing may be the inner skin of the royal family or the inner skin of the bamboo shoot.

According to another preferred embodiment of the present invention, the kill extract may contain coumaric acid (p-coumaric acid) or tricine (tricin) component.

According to another preferred embodiment of the present invention, the kill extract

i) acetylcholinesterase (AChE) inhibitory activity;

ii) antioxidant activity;

iii) lipid peroxide accumulation inhibitory activity; and

iv) inhibiting the production of inflammatory cytokines, including IL-1, IL-6, and TNF-α, and promoting the production of anti-inflammatory cytokines, including IL-10;

According to another preferred embodiment of the present invention, the cognitive dysfunction is mild cognitive impairment, attention deficit disorder (ADHD), depression, Alzheimer's disease, cerebrovascular dementia, senile dementia, Pick's disease and Creutzfeldt- It may be selected from the group consisting of Jacob's (Creutzfeldt-jakob's) disease.

The extract of the present invention, especially the ethanol extract of the kill, contains a high content of coumaric acid (p-coumaric acid) and tricin, and effectively suppresses the inflammatory response induced in microglia stimulated with LPS. Confirmed.

In addition, the extract of the present invention not only showed a memory enhancing effect in the Y-maze experiment and passive avoidance experiment using mice whose memory impairment was induced by scopolamine, but also showed acetylcholinesterase inhibitory activity, antioxidant activity, lipid peroxide accumulation Since it was confirmed that the inhibitory activity and the expression inhibitory activity of inflammatory biomarkers were shown, it can be usefully used in pharmaceutical compositions or health functional foods for the prevention or treatment of memory and cognitive dysfunction.

Figure 1 is (A, B) coumaric acid (p-coumaric acid), (C, D) tricin (tricin) and (E, F) caffeic acid (E, F) contained in porridge (inner bark) extract and bamboo leaf (leaf) extract Caffeic acid) content was confirmed.
Figure 2 is when the LPS-stimulated BV-2 microglia cells were killed and the extract was treated by concentration, (A) nitrite (NO, Nitrite), (B) prostaglandin E ₂ (PEG ₂ , Prostaglandin E ₂ ) This is data confirming the degree of generation of
Figure 3 is data confirming the expression level of (A) mRNA and (B) protein of iNOS when the LPS-stimulated BV-2 microglial cells were killed and the extract was treated by concentration.
Figure 4 is data confirming the protein expression level of COX-1 and COX-2 when the LPS-stimulated BV-2 microglia was killed and the extract was treated by concentration.
Figure 5 is when the LPS-stimulated BV-2 microglial cells were killed and the extract was treated by concentration, (A) TNF-α, (B) IL-1β, (C) IL-6 production and (D) these This is the data confirming the mRNA expression level of .
6 is data confirming the degree of inhibition of IκB-α degradation when LPS-stimulated BV-2 microglial cells were killed and the extract was treated by concentration.
Figure 7 is data confirming the acetylcholinesterase inhibitory ability of (A) killed methanol extract and (B) killed butanol extract.
8 is LPS-stimulated BV-2 microglial cells killed when the extract was treated by concentration, (A) nitrite production inhibitory activity and (B) data confirming the degree of cytotoxicity.
Figure 9 is data showing the results of a passive avoidance test (passive avoidance test) when the animal model of cognitive impairment disease induced by scopolamine was killed and treated with an ethanol extract.
10 is data showing the results of the Y-maze experiment when an animal model of cognitive impairment disease induced by scopolamine was killed and treated with an ethanol extract.
11 is data confirming the activity of antioxidant enzymes in the hippocampus and cerebral cortex when the animal model of cognitive impairment disease induced by scopolamine was killed and treated with an ethanol extract.
FIG. 11a is SOD (Superoxide Dismutase) activity, FIG. 11B is CAT (Catalase) activity, and FIG. 11C is GPx (Glutathione peroxidase) activity.
Figure 12 is data confirming the level of lipid peroxidation indicator MDA (Malondialdehyde) in the hippocampus and cerebral cortex when the animal model of cognitive impairment disease induced by scopolamine was killed and treated with ethanol extract .
Figure 13 is data confirming the degree of neuroinflammation in the hippocampus and cerebral cortex when the animal model of cognitive impairment disease induced by scopolamine was killed and treated with an ethanol extract.
FIG. 13a shows data confirming the level of TNF-α, FIG. 13b the level of IL-1β, FIG. 13c the level of IL-1, and FIG. 13d the level of IL-10.

Hereinafter, the present invention will be described in detail.

Composition for preventing or treating neuroinflammatory diseases

In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating neuroinflammatory diseases comprising an extract as an active ingredient.

From another aspect, the present invention relates to a health functional food composition for preventing or improving neuroinflammatory diseases comprising an extract as an active ingredient.

In the present invention, the killing may be the inner skin of the royal family or the inner skin of the bamboo shoot.

In the present invention, the extract may be prepared according to a conventional method known in the art, that is, under conventional temperature and pressure conditions, using a conventional solvent, preferably purified water (or water) , lower alcohols having 1 to 4 carbon atoms, glycerin, ethyl acetate, acetone, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol, hexane, and at least one selected from the group consisting of mixed solvents thereof Extracted using a solvent, more preferably extracted using water or ethanol (alcohol), but is not limited thereto.

In the present invention, the kill extract may contain a coumaric acid (p-coumaric acid) or tricine (tricin) component.

In a specific embodiment of the present invention, hot water, 30% ethanol (alcohol), 70% ethanol (alcohol), and 100% ethanol (alcohol) are prepared by using the inner bark (killer) or leaves (bamboo leaf) of king stem, bamboo shoots, and ojuk. An extract was prepared using As a result of comparing the contents of p-coumaric acid, tricin, and caffeic acid as active ingredients of each extract, as shown in Figure 1 and Table 2, in the case of the extract, bamboo leaf extract It was confirmed that the tricine content was significantly higher than that of In addition, coumarin acid was confirmed to be high in 100% ethanol extract and tricine in 70% ethanol extract.

In the present invention, the kill extract

i) inhibition of production of inflammatory mediators including NO, prostaglandin E ₂ , iNOS and COX-2;

ii) inhibition of production of inflammatory cytokines including IL-1β, IL-6 and TNF-α; and

iii) inhibition of IκB-α phosphorylation and degradation; can regulate neuroinflammation.

In another specific embodiment of the present invention, in order to confirm the neuroinflammation-regulating effect of the extract, NO production inhibitory effect, iNOS and COX-2 expression inhibitory effect, pro-inflammatory mediators IL-1β, IL-6 and TNF- α production inhibitory effect and NF-κB activity inhibitory effect (IκB-α degradation inhibitory effect) were confirmed.

As a result, the extract of the present invention inhibits LPS-induced NO production and prostaglandin E ₂ (FIG. 2), inhibits iNOS and COX-2 mRNA/protein expression (FIGS. 3 and 4), and IL-1β , It was confirmed that there is an effect of inhibiting the production of inflammatory cytokines including IL-6 and TNF-α (FIG. 5). In addition, it was confirmed that the extract of the present invention inhibits IκB-α degradation to block NF-κB signaling (FIG. 6).

In the present invention, the extract is preferably included in the total composition at a concentration of 10 to 250 μg/ml, preferably 10 to 200 μg/ml, and more preferably 10 to 150 μg/ml. When the extract is included at a concentration of less than 10 μg/ml, the effect of preventing or treating neuroinflammation may be insignificant, and when the extract is included at a concentration exceeding 250 μg/ml, cytotoxicity may occur (FIG. 7) .

In the present invention, "neuroinflammatory disease" is a general term for diseases that cause degenerative changes in nerve cells of the central nervous system and cause various symptoms, and representative neuroinflammatory diseases according to the present invention include multiple sclerosis, neuroblastoma, ischemic stroke, It may be any one selected from the group consisting of Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, Huntington's disease, Creutzfeldt-Jakob disease, post-traumatic stress disorder, depression, schizophrenia, or amyotrophic lateral sclerosis.

Composition for preventing or treating memory and cognitive dysfunction

In another aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of memory and cognitive dysfunction comprising an extract as an active ingredient.

In another aspect, the present invention relates to a health functional food composition for preventing or improving memory and cognitive dysfunction, comprising an extract as an active ingredient.

In the present invention, the killing may be the inner skin of the royal family or the inner skin of the bamboo shoot.

In the present invention, the extract may be prepared according to a conventional method known in the art, that is, under conventional temperature and pressure conditions, using a conventional solvent, preferably purified water (or water) , lower alcohols having 1 to 4 carbon atoms, glycerin, ethyl acetate, acetone, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol, hexane, and at least one selected from the group consisting of mixed solvents thereof Extracted using a solvent, more preferably extracted using water or ethanol, but is not limited thereto.

In the present invention, the kill extract may contain a coumaric acid (p-coumaric acid) or tricine (tricin) component.

According to another preferred embodiment of the present invention, the kill extract

i) acetylcholinesterase (AChE) inhibitory activity;

ii) antioxidant activity through SOD (Superoxide Dismutase), CAT (Catalase), GPxGlutathione peroxidase) enzyme activities;

iii) lipid peroxide accumulation inhibitory activity; and

iv) inhibiting the production of inflammatory cytokines, including IL-1, IL-6, and TNF-α, and promoting the production of anti-inflammatory cytokines, including IL-10;

In a specific embodiment of the present invention, in order to confirm the ability of the kill extract to improve memory or cognitive function, a passive avoidance test and a Y-maze test were performed, and acetylcholinesterase inhibitory effect, antioxidant effect, lipid peroxide The accumulation inhibitory effect and the inflammatory cytokine production effect were confirmed.

As shown in Figure 8, as a result of confirming the acetylcholinesterase inhibitory ability using the killed methanol or butanol fraction, it was confirmed that both the methanol fraction and the butanol fraction inhibit acetylcholinesterase activity.

In mice whose memory was impaired by scopolamine, after each treatment with the killed ethanol extract, a passive avoidance test was performed. As a result, as shown in FIG. confirmed that In addition, as shown in Figure 10, it was confirmed that the alternation behavior was significantly reduced by the ethanol extract in the alternation behavior experiment using the Y-maze experimental device.

In another specific embodiment of the present invention, when a mouse whose memory was impaired with scopolamine was killed and treated with an extract, the degree of oxidative stress was confirmed in the hippocampus and cerebral cortex, and as a result, the antioxidant enzyme SOD ( Superoxide Dismutase), CAT (Catalase), and GPxGlutathione peroxidase) activities were increased (FIG. 11a to FIG. 11c), and it was confirmed that the level of MDA (malondialdehyde), an indicator of lipid peroxidation, decreased (FIG. 12).

In addition, in order to confirm the neuroinflammation inhibitory effect in the hippocampus and cerebral cortex, as a result of checking the levels of TNF-α, IL-1β, IL-6 and IL-10, the inflammatory cytokines TNF-α, IL It was confirmed that the production of -1β and IL-6 decreased (FIGS. 13a to 13c), and the production of anti-inflammatory cytokine IL-10 increased (FIG. 13d).

In the present invention, the cognitive dysfunction is mild cognitive impairment, attention deficit disorder (ADHD), depression, Alzheimer's disease, cerebrovascular dementia, senile dementia, Pick's disease and Creutzfeldt-Jakob disease It may be selected from the group consisting of, but is not limited thereto.

The pharmaceutical composition of the present invention may be formulated and used in various forms according to conventional methods. For example, it can be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions and syrups, and can be formulated and used in the form of external preparations, suppositories and sterile injection solutions. Depending on each formulation, pharmaceutically acceptable carriers, excipients and diluents may be further included. In addition, it can be formulated and used in the form of external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and sterile injection solutions according to conventional methods.

The carriers, excipients and diluents include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, mineral oil and the like. When formulating or formulating the pharmaceutical composition, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose, etc. It is prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. there is. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

As used herein, the term "administration" means providing the pharmaceutical composition of the present invention to a subject by any suitable method. The pharmaceutical composition of the present invention is the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human thought by a researcher, veterinarian, physician or other clinician, that is, alleviation of the symptoms of the disease or disorder being treated It can be administered in a therapeutically effective amount, which is an amount that induces It is apparent to those skilled in the art that the therapeutically effective dosage and frequency of administration of the pharmaceutical composition of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, the patient's age, weight, and general health condition , gender and diet, administration time, route of administration and secretion rate of the composition, treatment period, can be adjusted according to various factors including drugs used simultaneously. The pharmaceutical composition of the present invention can be administered to a subject by various routes. For example, intravenous, intraperitoneal, intramuscular, intraarterial, buccal, intracardiac, intramedullary, intrathecal, transdermal, intestinal, subcutaneous, sublingual or topical administration may be administered, but is not limited thereto. The pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg/kg/day, and may be administered once a day or divided into several times.

The health functional food composition of the present invention may be used as a health functional food, food additive or dietary supplement. When using the extract of the present invention as a food additive, it may be appropriately used according to a conventional method, such as being added as it is or mixed with other foods or food ingredients.

In addition, the mixing amount of the health functional food composition may be appropriately changed according to the purpose of use (prevention, health or therapeutic treatment). As a specific example, when preparing food or beverage, the extract of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, for long-term intake for health and hygiene purposes or health control purposes, it may be added in an amount below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. there is.

There is no particular limitation on the type of food, but examples of food to which the extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, and ice cream. Dairy products, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, etc., include all health foods in a conventional sense.

When the health functional food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavoring agents or natural carbohydrates, like conventional beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; natural sweeteners such as dextrins and cyclodextrins; Synthetic sweeteners such as saccharin and aspartame may be used. The natural carbohydrate is included in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight, based on the total weight of the food composition of the present invention.

The health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , carbonation agents used in carbonated beverages, etc., and may include fruit flesh for the manufacture of natural fruit juice, fruit juice beverages and vegetable beverages, but are not limited thereto. These components may be used independently or in combination. The proportion of the above additives is not particularly limited, but is preferably included within the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

Hereinafter, the present invention will be described in more detail through examples.

These examples are only for exemplifying the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Preparation of Kyolyeo extract and confirmation of main active ingredient content

1-1: Manufacture of kill and bamboo leaf extract

In the present invention, as shown in Table 1 below, an extract was prepared using the inner skin (kill) or mouth (bamboo leaf) of ojok, kingdae, and bamboo shoots.

Sample type and extraction solvent hydrothermal 30% ethanol 70% ethanol 100% ethanol Oh Juk 152g 152g 77g 76g king 106.5g 106.5g 53g 53g bamboo shoots 90g 90g 90g 45g

The hot water extract, 30% (v/v) ethanol extract, 70% (v/v) ethanol extract, and 100% (v/v) ethanol extract were extracted under reflux under the following extraction conditions, and filtered using a 10 μm filter cloth. Next, each extract was prepared by concentration under reduced pressure.

The extraction conditions are as follows.

- Hot water: 4L of purified water, temperature 60℃, extraction time 16 hours

- 30% ethanol: 4L of 30% alcohol, temperature 60℃, extraction time 16 hours

- 70% ethanol: 4L of 70% alcohol, temperature 60℃, extraction time 16 hours

- 100% ethanol: 4L of 100% alcohol, temperature 60℃, extraction time 16 hours

1-2: Check active ingredient content

The contents of p-coumaric acid, tricin, and caffeic acid as active ingredients of each extract were compared.

active ingredient content p-coumaric acid
(μg/mg) Tricin
(μg/mg) Caffeic acid
(μg/mg) king
inner skin hydrothermal 0.1 0 0.457 30% alcohol 4.364 7.406 0.185 70% alcohol 4.987 20.58 0.127 100% alcohol 7.034 19.225 0.258 bamboo shoots
inner skin hydrothermal 0 0 0.4561 30% alcohol 2.217 5.327 0 70% alcohol 3.028 12.695 0 100% alcohol 4.002 8.890 0 king
leaf hydrothermal 1.557 0.325 0 30% alcohol 6.799 2.627 1.1477 70% alcohol 7.345 3.094 1.2637 100% alcohol 9.168 4.856 4.6300 Oh Juk
leaf hydrothermal 1.968 0.412 62.4704 30% alcohol 6.944 1.149 13.3346 70% alcohol 10.931 5.233 22.2112 100% alcohol 9.374 3.094 38.7871 bamboo shoots
leaf hydrothermal 1.371 0.298 0 30% alcohol 6.502 2.538 1.3804 70% alcohol 6.774 4.458 1.6943 100% alcohol 6.496 4.276 1.7734

As shown in Table 2 and FIG. 1, it was confirmed that the content of tricin was significantly higher than that of bamboo leaves in the case of the kill extract, and in particular, the p-coumaric acid in the kill extract of both king and bamboo shoots. It was found that tricin content was relatively high. In addition, coumaric acid (p-coumaric acid) was found to be high in the 100% alcohol extract and tricin (tricin) in the 70% alcohol extract. In the case of the kill extract, it was confirmed that the content of caffeic acid was insignificant.

Confirmation of inhibitory efficacy of inflammatory mediators by killing extract from LPS-stimulated microglia

2-1: Confirmation of NO generation suppression effect

In the present invention, in order to confirm the neuroinflammation inhibitory effect of the extract, the nitrite (NO) release inhibitory effect was measured, and the nitrite concentration was measured by a known method (Kim, JJ, et al. , Journal of Ethnopharmacology , 140: 213-221, 2012) to measure BV-2 cell viability.

BV-2 cells were cultured in DMEM medium supplemented with inactivated 5% FBS and 100 units/ml 1% penicilline/streptomycin under conditions of 5% CO2 and 95% O2 in a humidified environment.

Specifically, the BV-2 cells were plated at 1×10 ⁴ cells/well in a 96-well plate, and then the concentration of the 30% alcohol extract of the kill (Pygmy persimmon) extracted in Example 1 was 1 μg/ml, 10 ug/ml and 100 μg/ml were added. After 1 hour, LPS (200 ng/ml), a neuroinflammatory inducer, was treated, and then BV-2 cells were stimulated for 24 hours, and nitrite (NO) produced at this time was analyzed to have inhibitory effect in a concentration-dependent manner.

Nitrite (NO) was measured using a NO assay kit (Abcam) using a Griess reagent, and the measurement method was performed according to the kit's instructions. After mixing 100 μl of the cell culture medium and the grease reagent, the mixture was reacted for 10 minutes and observed at 540 nm absorbance, and the concentration was finally confirmed using a standard curve.

As a result, as shown in FIG. 2a, when BV-2 cells were treated with LPS, NO production was significantly increased compared to the control group, but it was confirmed that NO production stimulated by LPS was suppressed by treatment with the kill extract.

2-2: Prostaglandin E ₂ (PEG ₂ , Prostaglandin E ₂ ) production inhibition efficacy

In the present invention, in order to confirm the neuroinflammation inhibitory effect of the kill extract, the degree of prostaglandin production was measured. Prostaglandin is produced under the action of COX-1 and COX-2, and blood vessels are dilated by prostaglandin, and these stimuli stimulate nerve cells and cause pain.

Prostaglandin was analyzed using the Prostaglandin E2 Parameter assay kit (R&D SYSTEM), based on the manufacturer's instructions. Briefly, 100 μl of the cell culture was reacted with an antibody to compete with Horseradish Peroxidase (HRP)-labeled PGE2 and color development was induced. Absorbance was measured at 450 nm, and the concentration was finally confirmed using a standard curve.

As a result, as shown in FIG. 2b, it was confirmed that the production of prostaglandin, which was increased by LPS, was killed and the production decreased in a concentration-dependent manner.

Confirmation of inhibitory effect of iNOS, COX-1 and COX-2 expression by killing extract in LPS-stimulated microglia

In the present invention, the inhibitory effect of mRNA and protein expression of iNOS (inducible nitric oxide synthase), COX-1 (cyclooxygenase-1) and COX-2 (cyclooxygenase-2), which are inflammatory response mediators, was confirmed.

3-1: Check mRNA expression level

BV-2 cells were seeded in a 6-well plate at 5 × 10 ⁶ cells/well per well and cultured overnight, then killed to obtain concentrations of 1 μg/ml, 10 μg/ml and 100 μg/ml of the extract. added. After 1 hour, LPS (200ng/ml), a neuroinflammatory inducer, was treated, and BV-2 cells were stimulated for 6 hours, followed by TRIzol reagent (Invitrogen Life Technologies, USA) according to the manufacturer's manual. For RT (reverse transcription) PCR, the sample was made to have a concentration of 1 μg/μl, cDNA was synthesized according to the cDNA synthesis method of ReverTra Ace (TOYOBO, JAPAN), and each PCR reaction was performed using 2 μl of cDNA ( 0.5 μg/μl).

PCR primer sequences and reaction conditions Primer sequence (5'-3') order
number iNOS F: 5'-GAG GTA CTC AGC GTG CTC CA-3' One R: 5'-AGG GAG GAA AGG GAG AGA GG-3' 2 GAPDH F: 5'-ACC ACA GTC CAT GCC ATC AC-3' 3 R: 5'-CCA CCA CCC TGT TGC TGT AG-3' 4

PCR was performed using AccuPower®RocketPlex RT-PCR PreMix (BIONEER), denaturation (94 ℃, 5 minutes), 20 to 28 cycles of amplification (94 ℃ for 30 seconds, 50 to 57 ℃ for 1 minute and 72 ℃ 1 min) and extension (5 min at 72 °C), and the PCR amplicons were analyzed using a 1% agarose gel. GAPDH mRNA serves as an internal control for sample loading and mRNA integrity.

3-2: Check protein expression level

BV-2 cells were seeded in a 6-well plate at 5 × 10 ⁶ cells/well per well and cultured overnight, then killed to obtain concentrations of 1 μg/ml, 10 μg/ml and 100 μg/ml of the extract. added. After 1 hour, LPS (200 ng/ml), a neuroinflammatory factor, was treated, and BV-2 cells were stimulated for 18 hours, followed by protein isolation.

To identify proteins, cells were identified using a sample buffer (glycerol, 10% SDS, 0.5M Tris-Hcl). Equal amounts of protein were electrophoresed on 10% SDS-PAGE and then transferred from the gel to the membrane using 0.2 μm nitrocellulose (NC: GE Healthcare Life Sciences).

Anti-β-actin antibody, anti-COX-1 antibody, anti-COX-2 antibody (1:2000; Santa Cruz), and anti-iNOS antibody (1:2000; Cell Signaling) were treated as primary antibodies at 4°C. was reacted overnight. Then, after washing three or more times with PBS, horseradish peroxidase (HRP)-attached secondary antibody was treated and reacted at room temperature for 1 hour. After the reaction was completed, the membrane was reacted with an ECL detection kit or Femto (Thermo, USA), and then imaged using a luminescent image analyzer (LAS-3000, USA).

As a result, as shown in FIG. 3 , when LPS was treated in BV-2 cells, iNOS expression significantly increased compared to the control group, whereas iNOS mRNA and protein expression levels were suppressed when the kill extract was pretreated.

In addition, as shown in FIG. 4, when the BV-2 cells were treated with LPS, the COX-2 expression was significantly increased compared to the control group, while the COX-2 protein expression level was also suppressed when the kill extract was pre-treated. . On the other hand, no change in COX-1 protein expression was observed by the killi extract.

Confirmation of inhibitory effect of proinflammatory cytokine expression by killing extract in LPS-stimulated microglia

It is known that the increase in the production of inflammatory cytokines is involved in the development of inflammation in the central nervous system through the secretion of chemokines that promote the migration of leukocytes to the brain, the induction of adhesion molecules, and the destruction of the BBB (Blood Brain Barrier). Therefore, in the present invention, when the LPS-stimulated BV-2 cells were killed and the extract was treated at different concentrations, the production and mRNA expression of pro-inflammatory cytokines including IL-1β, IL-6, and TNF-α were inhibited. confirmed that it is.

BV-2 cells were cultured in the same manner as in Example 3-1, mRNA was isolated from the cells, and PCR was performed for each target using the synthesized cDNA as a template. In addition, the production levels of IL-1β, IL-6, and TNF-α were measured using each ELASA kit.

PCR primer sequences and reaction conditions Primer sequence (5'-3') order
number IL-6 F: 5'-TGA TGC ACT TGC AGA AAA CA-3' 5 R: 5'-ACC AGA GGA AAT TTT CAA TAG GC-3' 6 IL-1β F: 5'-TGT GAA ATG CCA CCT TTT GA-3' 7 R: 5'-GGT CAA AGG TTT GGA AGC AG-3' 8 TNF-α F: 5'-AGG GAG AGT GGT CAG GTT GC-3' 9 R: 5'-CAG CCT GGT CAC CAA ATC AG-3' 10 GAPDH F: 5'-ACC ACA GTC CAT GCC ATC AC-3' 3 R: 5'-CCA CCA CCC TGT TGC TGT AG-3' 4

As a result, as shown in FIG. 5, when BV-2 cells were stimulated with LPS, the levels of inflammatory cytokines such as TNF-α, IL-1β, and IL-6 increased, whereas the cytokines were killed in a concentration-dependent manner. It was confirmed that the gene expression of was decreased, and the cytokine concentration produced accordingly was also decreased. Through this, it was confirmed that the kill extract of the present invention inhibits the production of inflammatory cytokines stimulated by LPS, and it was confirmed that this inhibition is made in the transcriptional stage.

Confirmation of inhibitory effect on NF-κB activity by killing extract in LPS-stimulated microglia

LPS is known to regulate iNOS gene expression through NF-κB activation. Therefore, in order to determine whether the kill extract inhibits the activation of the NF-κB signaling pathway, it was confirmed whether the kill extract inhibits IκBα degradation in BV-2 cells induced by LPS.

BV-2 cells were dispensed into a 6-well plate at 5 × 10 ⁶ cells/well per well and cultured for 12 hours, then killed and the concentrations of the extracts were 1 μg/ml, 10 μg/ml and 100 μg/ml. added as much as possible. After 1 hour, LPS (200 ng/ml), a neuroinflammatory factor, was treated, and BV-2 cells were stimulated for 30 minutes. Then, proteins were isolated and western blotting was performed.

Protein separation and Western blotting were performed in the same manner as in Example 3-2, and anti-IκB-α antibody and anti-β-actin antibody (1:2000; Sigma-Aldrich, USA) were used as primary antibodies. .

As a result, as shown in FIG. 6 , it was confirmed that IκB-α degradation was significantly attenuated in LPS-stimulated BV-2 cells by the killi extract. From this, it can be inferred that the suppression of NO production and inflammatory cytokine production by the kill extract in LPS-stimulated BV-2 cells is due to blocking of the NF-κB or MPAK signaling system.

Confirmation of acetylcholinesterase inhibitory effect of Khalya extract

It has been reported that as the concentration of acetylcholine esterase (AChE) increases in the cerebral blood vessels, cholinergic neurotransmitters in nerve cells are deficient, which can cause memory and cognitive dysfunction, so measurement of AChE inhibitory activity can be used to prevent and treat dementia. Or it can be used as a tool for the development of drugs or health functional raw materials for cognitive enhancement. Therefore, in the present invention, the AChE inhibitory activity was measured using the extract or purified fraction.

The AChE inhibitory activity was performed by partially modifying the Ellman's coupled enzyme assay method. Specifically, in a 96-well plate, 170 μl of 100 mM phosphate buffer (pH 8), 20 μl of 2 mM DTNB (dithiobisnitrobenzoic acid), and 20 μl of kill extract were mixed, followed by 20 μl of AChE (0.25 U/buffer ml). was added and reacted at 37° C. for 10 minutes, and then 3.75 mM acetylcholine iodide substrate solution was added. After reacting the enzyme reaction solution at 37 ° C for 10 minutes, the absorbance was measured on a 410 nm UV-VIS microreader, and the absorbance (Ac) of the control group without substrate and sample and the test group By comparing the absorbance (As), the AChE inhibitory activity was calculated using Equation 1 below.

[Equation 1]

AChE inhibitory activity (%) = [1-(As/Ac)] X 100

Ac: absorbance of the control group, As: absorbance of the sample group

As a result, as shown in FIG. 7, it was confirmed that the bamboo leaf methanol extract and the bamboo leaf butanol (n-butanol) fraction extract reduced the activity of acetylcholinesterase. This means that the extract of the present invention can be used as a cognitive function improvement and dementia disease treatment.

Confirmation of inhibition of cytotoxicity and NO generation according to the concentration of the killi extract

BV-2 cell viability was measured according to a known method (Kim, JJ, et al. , Journal of Ethnopharmacology , 140:213-221, 2012) to measure the degree of cytotoxicity according to the concentration of the kill extract.

Cell culture and measurement of NO generation were performed in the same manner as in Example 2-1, and the concentration of the extract was 15.6 μg/ml, 31.25 μg/ml, 62.5 μg/ml, 125 μg/ml, and 250 μg of the extract. /mL and 500 μg/mL were added.

The cytotoxicity test of the kill extract was performed using the MTT measurement method. Specifically, after 24 hours of LPS stimulation, the supernatant was removed, and then DMEM medium containing MTT (500 μg/ml) was added to each well and cultured at 37° C. for 4 hours. After incubation, the supernatant was removed and DMSO (dimethyl sulfoxide); Compound Diluent/(1%) Vehicle control, Sigma, USA) was added to each well to dissolve formazan. The optical density was measured at 550 nm using a microplate reader (Tecan Trading AG, Switzerland).

As a result, as shown in FIG. 8, it was confirmed that the kill extract suppressed NO generation in a concentration-dependent manner up to a concentration of 125 μg/ml, and exhibited cytotoxicity at concentrations of 250 μg/ml and 500 μg/ml. Based on this, the concentration of the extract used in animal experiments was determined to be 100 mg/kg.

Confirmation of the cognitive function improvement effect of the kill extract in an animal model of cognitive impairment disease induced by scopolamine - Passive avoidance test

In the present invention, in order to confirm the effect of the extract on memory enhancement and prevention or treatment of cognitive dysfunction, the extract was administered to animal models of cognitive impairment caused by scopolamine.

Male C57BL/6 mice (25 g, 8 weeks old) were purchased from Daehan Biolink. They were divided into 4 groups and acclimated for 1 week under standard conditions, and then used in the experiment (temperature 22±2 °C, humidity 55±5%, 12 h-light/dark cycle, free food/water). All experiments were performed according to the guidelines of the Laboratory Animal Care Standards (NIH publication No. 85-23, revised 1985) and Konkuk University IACUC (Institutional Animal Care and Use Committee).

1) vehicle (untreated group, negative control group),

2) Scopolamine 2 mg/kg treatment group

3) Scopolamine 2 mg/kg + Killed ethanol extract 100 mg/kg treatment group

4) Scopolamine 2 mg/kg + tacrine (TAC) treatment group (positive control group)

All treatment groups were used by dissolving in physiological saline. Ethanol extract and tacrine were orally administered to mice every day at 100 mg/kg for 7 days before treatment with scopolamine, and 2 mg/kg of scopolamine was administered via intraperitoneal injection.

A passive avoidance experiment was conducted to evaluate memory, and for training and testing for passive avoidance behavior, a passive avoidance experiment system ( Active and passive avoidance system) were used. These devices have a light chamber (a bright room) and a dark chamber (a dark room), separated by computer-controlled doors. After putting the mouse in a bright room, light was applied to the dark room, and an electric current was passed through the mouse to remember it. The time until the mouse entered the dark room was defined as an acquisition trial.

When the mouse enters a dark room, the door automatically closes and a 0.5 mA electrical foot shock is given for 5 seconds. At this time, the transfer latency time (TLT), the time taken to avoid from a bright room to a dark room, was recorded and stored through a computer, and the electric shock and opening and closing of the door were controlled by the attached device system. The time it takes for the mouse to enter the dark room (latency time; retention time) was measured up to 300 seconds, and the longer the time it takes for the mouse to enter the dark room, the better the mouse's learning and memory.

As a result, as shown in FIG. 9, the time taken to avoid moving from a bright room (light chamber) to a dark room (dark chamber) by scopolamine treatment (TLT) was significantly reduced compared to the negative control group, but killing It was confirmed that TLT increased at a level similar to that of the tacrine-treated group by administration of the ethanol extract. This means that the ethanol extract of the present invention has the effect of improving cognitive function.

Confirmation of the spatial perception improvement effect of the kill extract in an animal model of cognitive impairment disease induced by scopolamine - Y-maze test

In the present invention, a Y-maze test was performed to confirm the effect of the extract on memory enhancement by the killifolia extract. In the same manner as in Example 8, the extract was administered to an animal model of cognitive impairment disease induced by scopolamine, and then the experiment was performed 30 minutes after scopolamine administration.

The apparatus of the Y-maze experiment is composed of three arms (each branch is designated as A, B, and C), and the length of each arm is 40 cm, the width is 12 cm, and the height is 30 cm. The angle of contact with the adjacent branch is 120 degrees, and all experimental devices are made of polyvinyl plastic. After carefully placing the mouse in the center of the maze and allowing it to move freely for 7 minutes, the branch the mouse entered was recorded, but only when the mouse fully entered the tail, and it was also recorded when the mouse repeatedly entered the branch again. The number and order of entering each branch were measured to evaluate the total number of entries (Total arm entry) and alteration behavior score (alteration, %) of the mouse. The change behavior score was defined as sequentially entering three branches, that is, ABC, BCA, CAB, etc., and was calculated by Equation 2 below.

[Equation 2]

% alteration = [total number of alterations]/[total number of times into arm - 2] X 100

As a result, as shown in Figure 10, the total number of entries (Total arm entry) of mice in each group was not significant. On the other hand, in the alternation behavior experiment using the Y-maze experimental device, the alternation behavior was significantly reduced after 30 minutes of scopolamine treatment, and in the mouse group pretreated with ethanol extract and tacrine, the reduction in alternation behavior was improved. confirmed that

Confirmation of oxidative stress improvement effect of kill extract in animal model of cognitive impairment disease induced by scopolamine

In the present invention, in order to confirm the efficacy of the ethanol extract to improve oxidative stress associated with memory impairment, antioxidant enzymes and lipid peroxidase activities were confirmed in the hippocampus and cerebral cortex of mice administered with the extract.

The experiment was performed in the same manner as in Example 8, and mice were sacrificed 60 minutes after administration of scopolamine to remove mouse brains.

After that, the hippocampus and cerebral cortex were carefully separated from the brain of the mouse, and antioxidant enzymes SOD (Superoxide Dismutase), CAT (Catalase), and GPx (Glutathione peroxidase) activities and MDA, which is an indicator of lipid peroxidation, were obtained. (Malondialdehyde) levels were measured.

To measure SOD, CAT, GPx and MDA activities, brain tissue homogeneous extracts were prepared using 10% (w/v) 0.1 M phosphate buffer, and superoxide dismutase detection kits (superoxide dismutase detection kits) were used. dismutase assay kit (Cell Biolabs, USA), catalase assay kit (Cell Biolabs, Canada), GPx colorimetric assay kit (BioVision, USA) and MDA detection kit (Lipid Peroxidation (MDA) Colorimetric/ It was measured using a Fluorometric Assay Kit; BioVision, USA). SOD was measured at 490 nm, CAT at 520 nm, GPx at 340 nm, and MDA at 410 nm.

As a result, as shown in Figures 11a to 11c, the antioxidant enzyme activity reduced by scopolamine administration, SOD, CAT and GPx activities were improved by the ethanol extract, and the same in the cerebral cortex and hippocampus of the brain was found to increase significantly.

Lipid peroxidation is caused by the involvement of oxygen free radicals generated by intrinsic as well as ex vivo factors. The brain has a high content of polyunsaturated fatty acids, which are prone to oxidative reactions, and consumes more oxygen in the body than other organs, making it susceptible to oxidative damage. It is known to degenerate.

As a result of confirming the level of lipid peroxidation by the kill extract, as shown in Figure 12, the increased lipid peroxidation level by scopolamine administration was reduced by the kill kill ethanol extract, and the same decrease in the cerebral cortex and hippocampus of the brain confirmed to be

Confirmation of neuroinflammation-reducing efficacy of the kill extract in an animal model of cognitive impairment disease induced by scopolamine

In the present invention, it was confirmed whether the neuroinflammation of an animal model of cognitive impairment disease induced by scopolamine is improved by the extract of sphagnum.

Using mouse hippocampus and cerebral cortex tissues extracted in Example 10, the levels of inflammatory cytokines TNF-α, IL-1β and IL-6 and anti-inflammatory cytokine IL-10 were measured. Confirmed. The degree of cytokine production was measured using each ELASA kit according to the product manual.

As a result, as shown in FIGS. 13a to 13c, it was confirmed that the levels of TNF-α, IL-1β and IL-6, which are inflammatory cytokines increased by scopolamine treatment, were significantly reduced by administration of the extract. In addition, as shown in Figure 13d, it was confirmed that IL-10, an anti-inflammatory cytokine, was decreased by scopolamine treatment, but increased by killing extract. That is, it was confirmed that neuroinflammation is improved by the extract of the present invention.

<110> GLOCAL Industry-Academic Cooperation Foundation Konkuk Universit <120> COMPOSITION FOR THE PREVENTION OR TREATMENT OFNEURODEGENERATIVE DISEASE OR COGNITIVE DYSFUNCTIONCOMPRISING BAMBOO INNER BARK EXTRACT <130> PDPC214305k01 <150> KR 10-2021-0083089 <151> 2021-06-25 <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> artificial sequence <220> <223> iNOS_forward primer <400> 1 gaggtactca gcgtgctcca 20 <210> 2 <211> 20 <212> DNA <213> artificial sequence <220> <223> iNOS_reverse primer <400> 2 agggaggaaa gggagagagg 20 <210> 3 <211> 20 <212> DNA <213> artificial sequence <220> <223> GAPDH_forward primer <400> 3 accacagtcc atgccatcac 20 <210> 4 <211> 20 <212> DNA <213> artificial sequence <220> <223> GAPDH_reverse primer <400> 4 ccaccaccct gttgctgtag 20 <210> 5 <211> 20 <212> DNA <213> artificial sequence <220> <223> IL-6_forward primer <400> 5 tgatgcactt gcagaaaaca 20 <210> 6 <211> 23 <212> DNA <213> artificial sequence <220> <223> IL-6_reverse primer <400> 6 accagaggaa attttcaata ggc 23 <210> 7 <211> 20 <212> DNA <213> artificial sequence <220> <223> IL-1b_forward primer <400> 7 tgtgaaatgc caccttttga 20 <210> 8 <211> 20 <212> DNA <213> artificial sequence <220> <223> IL-1b_reverse primer <400> 8 ggtcaaaggt ttggaagcag 20 <210> 9 <211> 20 <212> DNA <213> artificial sequence <220> <223> TNF-a_forward primer <400> 9 aggggagagtg gtcaggttgc 20 <210> 10 <211> 20 <212> DNA <213> artificial sequence <220> <223> TNF-a_reverse primer <400> 10 cagcctggtc accaaatcag 20

Claims (14)

A pharmaceutical composition for preventing or treating neuroinflammatory diseases comprising an extract as an active ingredient.
The pharmaceutical composition for preventing or treating neuroinflammatory diseases according to claim 1, wherein the extract is extracted with at least one solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.
The pharmaceutical composition for preventing or treating neuroinflammatory diseases according to claim 1, wherein the killing is the inner skin of the royal family or the inner skin of the bamboo shoot.
The pharmaceutical composition for the prevention or treatment of neuroinflammatory diseases according to claim 1, wherein the porridge extract contains p-coumaric acid or tricin.
The method of claim 1, wherein the kill extract
i) inhibition of production of inflammatory mediators including NO, prostaglandin E ₂ , iNOS and COX-2;
ii) inhibition of production of inflammatory cytokines including IL-1β, IL-6 and TNF-α; and
iii) inhibition of phosphorylation and degradation of IκB-α; a pharmaceutical composition for the prevention or treatment of neuroinflammatory diseases characterized by regulating neuroinflammation through.
The method of claim 1, wherein the neuroinflammatory disease is multiple sclerosis, neuroblastoma, ischemic stroke, Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, Huntington's disease, Creutzfeldt-Jakob disease, post-traumatic stress disorder, depression, schizophrenia or amyotrophic lateral sclerosis A pharmaceutical composition for the prevention or treatment of neuroinflammatory diseases, characterized in that any one selected from the group consisting of.
A composition for a health functional food for preventing or improving neuroinflammatory diseases comprising an extract as an active ingredient.
A pharmaceutical composition for preventing or treating memory and cognitive dysfunction comprising an extract as an active ingredient.
[Claim 9] The pharmaceutical composition for preventing or treating memory and cognitive dysfunction according to claim 8, wherein the extract is extracted with at least one solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.
[Claim 9] The pharmaceutical composition for the prevention or treatment of memory and cognitive dysfunction according to claim 8, wherein the killing is the inner skin of the royal family or the inner skin of the bamboo shoot.
[Claim 9] The pharmaceutical composition for preventing or treating memory and cognitive dysfunction according to claim 8, wherein the porridge extract contains p-coumaric acid or tricin.
The method of claim 8, wherein the kill extract
i) acetylcholinesterase (AChE) inhibitory activity;
ii) antioxidant activity;
iii) lipid peroxide accumulation inhibitory activity; and
iv) improving memory and cognitive function through inhibitory activity of inflammatory cytokines, including IL-1, IL-6, and TNF-α, and promoting activity of anti-inflammatory cytokines, including IL-10; A pharmaceutical composition for preventing or treating memory and cognitive dysfunction, characterized in that.
The method of claim 8, wherein the memory and cognitive function disorders include mild cognitive impairment, attention deficit disorder (ADHD), depression, Alzheimer's disease, cerebrovascular dementia, senile dementia, Pick's disease, and Creutzfeldt-Jacob. jakob) a pharmaceutical composition for preventing or treating memory and cognitive dysfunction, characterized in that any one selected from the group consisting of diseases.
A composition for health functional food for preventing or improving memory and cognitive dysfunction, comprising an extract as an active ingredient.

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