KR101201449B1 - Composition comprising the extract of grapefruit for preventing and improvement cognitive dysfunction and health supplementary food containing the same - Google Patents

Abstract

The present invention relates to a food composition for preventing and improving cognitive impairment containing grapefruit extract as an active ingredient, and a health supplement comprising the same. The food composition and dietary supplement for preventing and improving cognitive dysfunction of the present invention, such as Alzheimer's dementia, by effectively inhibiting the activity of acetylcholine esterase (AChE), an enzyme that degrades acetylcholine, a neurotransmitter. Cognitive dysfunction can be prevented and improved at the same time.

Description

Composition comprising the extract of grapefruit for preventing and improvement cognitive dysfunction and health supplementary food containing the same}

The present invention relates to a food composition for preventing and improving cognitive dysfunction and a health supplement comprising the same.

The development of medical technology and the improvement of living standards have extended the average life span of humans nearly twice in the last half century, which has resulted in a rapid increase in the ratio of the elderly to the total population. According to the 2005 data published by the National Statistical Office, the average life span of Koreans is 78.5 years old, and in 2000, about 7% of the population over 65 years old are entering the aging society rapidly. In addition, Korea is expected to enter an aging society in 2018 and an aging society where the proportion of the elderly population is over 20% in 2026. The old age support ratio (number of people over 65 / 15-64), which was 12.1% in 2004, is expected to increase to 21.3% in 2020 and 35.7% in 2030. If so, one elderly person per 6.8 in 2010, one elderly person per 4.7 in 2020, and one elderly person per 2.8 in 2030. This implies that the country is more likely to have a social burden due to lower economic growth, lower national competitiveness and deeper generational conflict.

Reflecting these social outlooks and reality, in recent years, attention has been focused on diseases that affect the elderly a lot. In particular, dementia is significantly higher in older adults, and it is known that the incidence rate is about 20% in elderly people aged 75 to 84 years and about 50% in elderly people aged 85 or older. It is becoming a serious social concern because it must be absolutely dependent. In addition, the economic loss due to dementia is increasing, and considering all the factors such as working loss, the social cost of dementia treatment in Korea is estimated to be between 3.4 trillion and 4.4 trillion won (2004 National Health Insurance Management Survey data). In the United States, the cost of dementia treatment is estimated to be between $ 100 and $ 120 billion annually. In other words, as society gradually ages, senile dementia has emerged as the biggest health problem that humankind has to solve in the 21st century. Accordingly, there is an increasing demand for the development of functional substances and foods capable of preventing and treating cognitive impairment including dementia.

Alzheimer's disease is a neurodegenerative disease that leads to progressive loss of cognitive and memory abilities associated with senile disease and neuronal loss. It is a neurotransmitter of the patient, acetylcholine, glutamic, neuropeptide and Neurotoxicity due to intracellular brain accumulation of β-amyloid protein, which induces functional disruption of monoamines and sustained destruction of neurons by oxidative stress, is presumed to be a major cause (Tanzi, RE et. al., Neuron, 2004, 43, 385-396). In modern society, the increase of degenerative brain diseases such as senile dementia has become a serious social problem due to the rapid increase of the elderly population, but until now, no effective medicines or treatments have been developed for the prevention and treatment of this disease. There is a situation. Accordingly, many studies have been conducted to develop medicines or functional foods for the treatment of dementia diseases. As a result, recently, the activity of acetylcholinesterase, an enzyme that degrades acetylcholine, a neurotransmitter Acetylcholinesterase inhibitors that increase brain acetylcholine have been developed and marketed. Typical drugs include tacrine, donepezil, COGNEX and galantamine, and rivastigmine, which are FDA approved drugs (Desai, AK et al. Neurology 64). , 34-39. 2005). However, most of these acetylcholinesterase inhibitors are very expensive, exhibit cholinergic side effects of peripheral nerves, have short half-lives, and have serious side effects such as hepatotoxicity, vomiting and loss of appetite (Christie, JE). et al., Br. J. Psychiatry 1981, 138, 46-50). In particular, 9-amino-1,2,3,4-tetrahydroacridine (THA), an active ingredient of COGNEX, has been shown to improve perceptual ability upon oral administration (Summers, WK, et al., New Engl J Med 1986 315, 1241-1246), tremors, dizziness, liver toxicity and other serious side effects have not been widely used. In addition, tacrine is characterized by hepatotoxicity in about half of the patients administered, and donepezil causes gastrointestinal disorders such as nausea, vomiting, diarrhea and constipation, and galantamine causes side effects such as nausea and vomiting, making it difficult to use for a long time.

Meanwhile, grapefruit (grapefruit) is a fruit of the citrus grapefruit tree, and its origin is Jamaica of the West Indies. The shape is round like a tangerine, the diameter is 10-15 cm, the outer skin is leather, the surface is smooth and yellow. In addition, the cuticle is thin and soft, and the flesh is pale yellow, rich in juice, sweet and strong in taste. These grapefruits are known to have the effect of inhibiting sebum control and inflammation exacerbation by acne while preventing sebum control and bacterial infection. However, there has been no report on the prevention and treatment of cognitive dysfunction such as Alzheimer's dementia on grapefruit extract.

The inventors of the present invention while researching to develop a composition for the prevention and improvement of cognitive dysfunction that can cause the treatment of dementia without side effects, the hydrothermal extract, organic solvent extract extracted from grapefruit is acetylcholine esterase (AChE) It has been confirmed that there is an effect of specifically inhibiting the activity of can be used for the prevention and improvement of cognitive dysfunction including Alzheimer's dementia.

It is an object of the present invention to provide a food composition for preventing and improving cognitive dysfunction, which is excellent in preventing and improving cognitive dysfunction related diseases such as forgetfulness and dementia, and a health supplement comprising the same.

The present invention for achieving the above object provides a food composition for preventing and improving cognitive impairment containing grapefruit extract as an active ingredient.

In addition, the grapefruit extract provides a food composition for preventing and improving cognitive dysfunction that is obtained by hot water extraction of grapefruit.

In addition, grapefruit extract provides a food composition for preventing and improving cognitive dysfunction that is extracted from a solvent selected from one or more alcoholic solvents selected from the group consisting of methanol, ethanol, propanol and butanol.

In addition, the grapefruit extract provides a food composition for preventing and improving cognitive dysfunction that is extracted from a solvent selected from one or more organic solvents selected from the group consisting of acetone, chloroform, methylene chloride, ether, ethyl acetate and hexane.

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The present invention also provides a dietary supplement comprising the food composition for preventing and improving the cognitive dysfunction.

In addition, it provides a dietary supplement comprising 0.05 to 5% by weight of the total weight of the food composition for preventing and improving cognitive dysfunction.

In addition, it provides a dietary supplement comprising a food composition for preventing and improving cognitive impairment of 0.05 to 5% by weight and 95 to 99.95% by weight of fermented milk.

In addition, the main ingredient of the fermented milk provides a dietary supplement, characterized in that choline fortified crude oil.

In addition, choline fortified crude oil provides a dietary supplement that contains 0.01 to 0.03% by weight relative to the total weight of crude oil.

According to the present invention, by inhibiting the activity of acetylcholine esterase (AChE), an enzyme that degrades acetylcholine, a neurotransmitter, it prevents cognitive dysfunction related diseases such as Alzheimer's dementia, and at the same time It is possible to provide a food composition for preventing and improving cognitive impairment having an excellent effect of improving a disorder, and a health supplement comprising the same.

1 is a flow chart schematically showing the manufacturing process of fermented milk for preventing and improving cognitive impairment according to an embodiment of the present invention.
Figure 2 shows the graph showing the enzyme activity inhibition of grapefruit extract according to an embodiment of the present invention.
Figure 3 shows a graph showing the memory enhancement effect of the grapefruit extract according to an embodiment of the present invention.
Figure 4 shows the grapefruit extract concentration in the fermented milk added grapefruit extract according to an embodiment of the present invention.
Figure 5 shows a graph showing the memory enhancement effect of the fermented milk added grapefruit extract according to an embodiment of the present invention.

The present invention relates to a food composition for preventing and improving cognitive dysfunction comprising a grapefruit extract as an active ingredient.

Grapefruit extract according to the present invention can be obtained by extracting grapefruit with water or an organic solvent, or a mixed solvent of water and organic solvent, the organic solvent is lower alcohol, acetone, chloroform, methylene chloride, ether, ethyl acetate, hexane, etc. Can be illustrated. Lower alcohols include methanol, ethanol, propanol and butanol, of which ethanol is most preferred.

As specific conditions for extracting grapefruit using water as a solvent, 5 to 15 times the weight of grapefruit, preferably 10 times the weight of grapefruit is added to the finely pulverized grapefruit and 25 to 100 ° C, preferably 70 to 95 ° C. Extraction for 1 to 72 hours, preferably 4 to 24 hours at a temperature and then filtered to prepare a hydrothermal extract of grapefruit.

When grapefruit is extracted using at least one alcoholic solvent selected from the group consisting of methanol, ethanol, propanol and butanol or at least one organic solvent selected from the group consisting of acetone, chloroform, methylene chloride, ether, ethyl acetate and hexane To the finely pulverized grapefruit is added 1 to 10 times, preferably 5 times the alcoholic solvent or the organic solvent of the weight of grapefruit and 1 to 72 hours, preferably 4 to 24 hours at a temperature of room temperature to 60 ℃ After extracting with stirring for a while, the extract obtained by filtration or centrifugation may be concentrated under reduced pressure to prepare a grapefruit extract according to the present invention.

In the extraction methods, the extraction process may be repeated two or more times as necessary, and the extract obtained after filtration or centrifugation may be used as it is, or may be dried under reduced pressure and / or lyophilized to obtain a powder form. .

The food composition for preventing and improving cognitive dysfunction according to the present invention may include the grapefruit extract alone, or may further include an extract of a pharmaceutically acceptable herbal medicine to enhance the cognitive dysfunction prevention and improvement effect. have. In this case, after the extract of the herbal medicines prepared according to the extraction method is added to the composition according to the invention or grapefruit and other herbal medicines may be included in the composition extract obtained by the extraction by the above method. Herbal medicines that may be added to the food composition for preventing and improving cognitive dysfunction of the present invention, specifically, Chrysanthemi flos, licorice (Glycyrrhizae radix), mistletoe (Viscum album), Cassiae semen, Cinnamonum (Cinnamonum) zeylanicum, Spatholubus suberectus, Red pepper herb (Hypericum erectum), Wolfberry (Lycii fructus), Gujeolcho (Chrysanthemum zawadskii), Locust (Raphanus sativus Linne), Elm (Ulmus davidiana), Ivy tricus (cuspidatasus) Rhei rhizoma, Chaenomeles sinensis, Cortex moutan, Myrrh (Commiphora myrrha), White egg (Sinapis alba L.), Butler (Typha latifolia), Loquat (Eriobotrya japonica Lindley), Betel nut (Areca catechu) Linne, Gemmingia chinensis, Sophora subprostrata, Sanzoin (Zizyphi spinosi Semen), Rhododendron yedoense, Morus alba Linne, Dendrobium moniliforme, Pine needles (Pinus densiflora) (Caesalpinia Sappan), Ambush (Cynomorium songaricum Ruprecht), Acacia Leaf (Robinia pseudoacacia), Night-gate Leaf (Lespedeza cuneata), Morus japonicus Muell.Arg, Schizandrae fructus, Broiler (Lycii radicis Cortex) Ginkgo (Gingko biloba), Ginseng (Panax ginseng), Peony (Paeoniae radix), Straw Sprout (Agrimonia pilosa), Brier (Rosa multiflora), Changza (Xanthium strumarium L.), Creation (Atractylodis rhizoma alba) Gastrodiae rhizoma, Lespedeza cyrtobotrya, Xanthium strumarium, Alpinia katsumadai Hayata, Cypress (Thuja rientalis L.), Cuscutae semem, Octagonal (Chinese star Anise), Cypress (Chinese star Anise) Chamaecyparis obtusa, Cyperus rotundus L., Corydalis (Corydalis turtschaninovii), Golden (Scutellariae radix), Laurel (Coptidis rhizoma), Yellow white (Phellodendri cortex), Yuri (Corylopsis coreana), Citron seed, Grape seed, Grape seed Sunflower Seed Extract, Majoran ), Cypress, Pine scotch, Basil, Rosemary, Tea tree and Eucalyptus. In particular, one or more herbal extracts selected from the group consisting of gujeolcho, Chinese quince, sagan, pine needles, eucalyptus, wild boil, straw sand, cypress, grape seed, rhododendron, yellow white and hyacinth are added alone or in combination of two or more. It is preferable because it is excellent in the cognitive function improving effect.

As a result of animal experiments, mice treated with the grapefruit extract or the mixture of grapefruit extract and other herbal extracts according to the present invention showed a marked dementia prevention effect and a cognitive function enhancing effect, but did not show acute toxicity and chronic toxicity. There were no adverse effects on liver function.

Grapefruit extract prepared by the method according to the invention or a composition containing it as an active ingredient inhibits the activity of acetylcholinesterase and thus inhibits the decomposition of acetylcholine in the body. Therefore, the food composition for preventing and improving cognitive dysfunction of the present invention containing a grapefruit extract as an active ingredient can be used for the purpose of preventing and improving diseases caused by the lack of acetylcholine in the body. Such diseases include dementia (Terry AV et al., J Pharmacol Exp Ther 2003, 306, 821-827; Giacobini E. Cholinesterases Neurochem Res 2003, 28, 515-522; Auld DS et al., Prog Neurobiol 2002, 68 , 209-245), memory loss (Myhrer T. Brain Res Rev 2003, 41, 268-287; Graves L. et al., Trends Neurosci 2001, 24, 237-243), myasthenia gravis (Levinson AI et al., Immunol Res 2003, 27, 399-408; Spiess JL et al., J Pain Symptom Manage 2003, 26, 684-686; Fotiou F. et al., J Neuropsychiatry Clin Neurosci 2000, 12, 514-515) and glaucoma ( Millard CB et al., 1995, 64, 1909-1918; Hyndiuk RA et al., Lens Eye Toxic Res 1992, 9, 331-350). In addition, the present invention includes a grapefruit extract as an active ingredient, harmless to the human body, has a simple manufacturing method, easy to purchase the raw material and low cost, it can be commercialized in large quantities.

The present invention can also provide a health supplement with a memory enhancing effect, including the food composition for preventing and improving the cognitive dysfunction. Foods to which the composition according to the present invention can be added for the purpose of preventing and improving cognitive dysfunction of the present invention, that is, providing a health supplement food having an effect of improving memory, include, for example, various fermented milks, beverages, confectionary, breads, Noodles, seasonings and vitamin complexes. Among them, dairy products, especially fermented milk, are not only effective in intestinal intestinal action, anticancer action, prevention of constipation and diarrhea, and stimulation of digestion and absorption of nutrients, but also contain a lot of protein and vitamin B. Since it helps to secrete hormones, the health supplement prepared by adding the grapefruit extract containing composition according to the present invention to fermented milk is preferable because it increases the effect of preventing and improving cognitive dysfunction, that is, memory. Fermented milk is the same as most of the nutritional ingredients mainly derived from raw milk compared to the general milk, but the difference in digestion improvement effect and lactic acid production effect on the human body, the digestion rate is known to be about twice the milk. In particular, in the case of fermented milk made from crude oil with a high content of choline (hereinafter referred to as 'choline fortified milk'), the main component of the brain and nervous system, choline, grapefruit extract and the components of fermented milk, It is highly desirable because it further enhances the effect of preventing and improving cognitive dysfunction of nutraceuticals. The choline fortified milk is preferably contained in the content of choline 0.01% by weight or more, and more specifically refers to milk containing choline 0.01 to 0.03% by weight relative to the total weight of crude oil.

The dietary supplement is easily added by adding the above-mentioned concentrate or dry powder of the present invention to the manufacturing process of the base food or by adding the process of adding the composition of the present invention after the preparation of the base food. You can get it. At this time, various flavors and other additives may be added as necessary.

In the dietary supplement of the present invention, the content of the food composition for preventing and improving cognitive dysfunction, which contains the grapefruit extract as an active ingredient, varies depending on the type and preference of the food, but the prevention effect of the grapefruit extract against cognitive dysfunction. And it is preferable to contain in an amount of 0.05 to 5% by weight of the total weight of the food in order to fully utilize the improvement effect. When the content of the composition is less than 0.05% by weight, the efficacy of the composition contained in the food is insignificant, and when the content is more than 5% by weight is not economical because the increase in efficacy is not high compared to the increased content.

As an example of the dietary supplement according to the present invention will be described a method for producing fermented milk comprising a food composition for preventing and improving cognitive impairment of the present invention.

First, prepare crude oil. Crude oil is not particularly limited, but in order to further enhance the cognitive impairment improvement effect and memory enhancement effect according to the present invention, crude oil having a high content of choline, which plays a key role in brain development as an essential nutrient (hereinafter, 'choline-enriched crude oil) It is desirable to prepare a '). Choline-enriched crude oil is either from artificially added choline to crude milk or from choline milk protected with ruminant protection technology (hereinafter referred to as 'ruminine protective choline') in milking cows and obtained by squeezing milk directly. It is suitable for this fortified milk, but due to the recent consumer's rejection of food additives, it is preferable to feed natural dietary choline fortified with milk containing rumen protective choline. The choline fortified crude oil is better the greater the content of choline, but preferably contains 0.01 to 0.03% by weight relative to the total weight of the crude oil. Crude oil containing less than 0.01% by weight of choline has little effect of choline, and when choline content exceeds 0.03% by weight, the production cost is high and economical is low.

Then, the prepared crude oil, especially choline fortified crude oil, is homogenized at a homogeneous pressure and sterilized. At this time, the homogeneous pressure is preferably 150 ~ 200bar, sterilization for 3 minutes to 10 minutes at 80 ℃ ~ 110 ℃. After sterilizing crude oil, it is preferable to quench it at the temperature of 37 degreeC-45 degreeC.

And inoculate lactic acid seed to sterilized crude oil for 5-8 hours Incubate. At this time, the lactic acid seed is not particularly limited as long as lactic acid bacteria, but it is preferable to use a mixture of L.bulgaricus and S.thermophilus in a ratio of about 1: 1. After the lactic acid bacteria culture is completed, it is preferable to terminate the culture at an appropriate acidity and cool rapidly with ice or the like.

Then, by adding a composition containing the grapefruit extract as an active ingredient to prepare a fermented milk for preventing and improving cognitive impairment according to the present invention.

According to the fermented milk production method for preventing and improving cognitive impairment of the present invention, it is possible to provide a functional fermented milk excellent in inhibiting effect of acetylcholinesterase and excellent memory enhancement effect.

Hereinafter, examples of the present invention will be described. The following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

<Examples>

Example  1-1: grapefruit Heat number  Preparation of extract

2 liters of distilled water was added to 200 g of dried and pulverized grapefruit, and hot water was repeatedly extracted twice at 90 ° C. to 95 ° C. for 6 hours. The supernatant obtained by centrifuging the extract at 10000 × g for 30 minutes was lyophilized to obtain 65 g of grapefruit hydrothermal extract of the present invention.

Example  1-2: Preparation of Ethanol Extract of Grapefruit

2 g of 99.9% ethanol was added to 200 g of dried and pulverized grapefruit, and extracted twice at room temperature for 24 hours. The filtrate obtained by filtration was concentrated under reduced pressure to obtain 70 g of grapefruit ethanol extract.

Example  2: Preparation of Fermented Milk Containing Grapefruit Extract

First, 20 kg of crude oil containing 5 g of choline was prepared. Each crude oil was preheated to 60 ° C., and the preheated crude oil was homogenized to a homogeneous pressure of 180 bar. After the homogenization was complete, the crude oil was sterilized for 5 minutes at 95 ℃. After cooling to 42 ° C., L.bulgaricus and S.thermophilus were inoculated with lactic acid bacteria mixed at a weight ratio of 1: 1, for 7.5 hours. The culture was carried out. After completion of the lactic acid bacteria culture, the culture was terminated at pH 0.8 and quenched with ice. Then, after the culture solution was stirred, the fermented milk according to the present invention (Examples 2-1 to 3) was completed by adding the grapefruit extract obtained in Example 1-1 at 0.1% by weight, 0.5% by weight and 1% by weight, respectively. .

Table 1 shows the formulation for the fermented milk according to the present invention.

Crude oil or
Choline Crude Oil (%) grapefruit
extract(%) Lactobacillus(%)
(L.bulgaricus and S.thermophilus) Sum(%) Control Non-choline crude oil
99.999 0 0.001 100 Comparative example Choline Crude Oil
99.999 0 0.001 100 Example 2-1 Choline Crude Oil
99.899 0.1 0.001 100 Example 2-2 Choline Crude Oil
99.499 0.5 0.001 100 Example 2-3 Choline Crude Oil
98.999 One 0.001 100

And, Table 2 shows the results of the production of fermented milk according to the present invention.

Control Comparative example Example 2-1 Example 2-2 Example 2-3 Incubation temperature 42 ℃ 42 ℃ 42 ℃ 42 ℃ 42 ℃ Incubation time 7H 30M 7H 30M 7H 30M 7H 30M 7H 30M Acidity 0.79 0.82 0.82 0.82 0.82 pH (17 ℃) 4.46 4.40 4.40 4.40 4.40

Experimental Example  1: of grapefruit extract Acetylcholinesterase  Inhibitory activity in vitro )

Acetylcholinesterase inhibitory activity against grapefruit extract of the present invention is described in Ellman GL et al. Biochem Pharmacol 1961, 7, 88-95 using acetylcholinesterase (Sigma C5021) present in the red blood cell membrane. Measurement was made using the presented method. Briefly, 10 µl of grapefruit extract and 10 µl of enzyme solution (0.5 U / ml acetylcholinese) at each concentration obtained in Example 1-1 were prepared in 450 µl of activity measuring buffer (50 mM Tris-HCl buffer, pH 8.2). Terase), mixed well, and left at room temperature for 5 minutes, and then 15 µl of 50 mM acetylthiocholine and 15 µl of 10 mM 5,5'-Dithio-Bis (2-Nitrobenzoic Acid) (DTNB) were added thereto. After the enzyme reaction was carried out at 37 ℃ and the absorbance was measured at a wavelength of 405nm, the enzyme activity inhibition of the extract was obtained from the following formula.

% Inhibition of enzymatic activity of the extract = {1-[(At-Ao) / (Ct-Co)]} x 100

In the above formula, Ao is the absorbance value measured immediately after the addition of grapefruit extract, At is the absorbance value measured after the reaction for t minutes after adding grapefruit extract, and Co is the absorbance value measured immediately after adding buffer instead of grapefruit extract as a control. And Ct represents absorbance values measured after adding buffer and reacting for t minutes. As a result of measuring acetylcholinesterase inhibitory activity, it was possible to obtain a graph of enzyme activity inhibition of grapefruit extract as shown in FIG. 2, and the IC ₅₀ value of grapefruit extract was about 0.5 μg / ml, and 5 μg / ml At the above concentration, the activity of the enzyme was suppressed almost 100%.

Experimental Example  2: Toxicity of Grapefruit Extract

In order to investigate the acute toxicity of the grapefruit extract obtained in Example 1-1, mortality, gross lesions and histologic lesions were observed 24 hours after single oral administration at a dose of up to 2 g / kg to ICR mice. Since no single death was observed in the experimental group, the LD ₅₀ value of grapefruit extract by oral administration was estimated to be greater than 2 g / kg. In addition, as a result of observing the gross lesions, the grapefruit extract did not show any difference from the control group not administered grapefruit extract, and the gross lesions were not observed. Therefore, the grapefruit extract is relatively safe in terms of side effects and acute toxicity when administered to animals. As such, it can be said to be an excellent material that can be safely used without substantial problems even in large doses.

Experimental Example  3: of grapefruit extract In vivo  effect( in vivo )

In order to investigate whether grapefruit extract shows dementia and cognitive dysfunction in vivo, memory loss was induced with scopolamine in ICR mice. A grapefruit extract with high inhibitory capacity was administered to observe the response to the passive avoidance test.

To measure the memory-promoting effect using the passive avoidance test, a shuttle box with a middle partition and divided into light and dark rooms left and right was used. One group of ICR (ICR) mice was orally administered the grapefruit extract (60 mg / kg) of Example 1-1, and the other group was administered only physiological saline as a solvent. After the scopolamine (1mg / ㎏) was injected into the abdominal cavity to the group that caused memory loss was performed passive avoidance test 30 minutes after scopolamine administration. The test animal is placed in a cubicle in a box with an electric grit attached to the floor (19L x 9W x 10.875H, Columbus Instruments, Model PACS-30, USA), and connected to a cubic door (3H x) with 1500 Lux lighting. 2.625W). At this point, the ICR mouse looks around the room and moves to the illuminated cubicle, which automatically closes. When the mouse was moved to the compartment room (B), the mouse was given a foot shock by passing a current of 1 mA for 5 seconds while the light was turned off. Then, the mouse remembers the relationship between the dark room and the sole shock, and after 24 hours, he puts it in the partition room A, and hesitates to move to the partition room B even if it lights up. At this time, the dwell time was measured by comparing the control group administered with the solvent and the group administered with the grapefruit extract. Figure 3 is a graph showing the memory enhancement effect of the grapefruit extract, the avoidance time of the normal group was 138.7 ± 11.9 seconds, the memory is excellent, on the contrary, when the scopolamine administered to give a memory impairment 22.7 ± 3.4 At first, memory was greatly reduced. However, when the grapefruit extract was administered followed by scopolamine, the avoidance time was 129.5 ± 10.3 seconds to confirm the inhibitory effect on memory loss.

Experimental Example  4: Grapefruit Extract Concentration and Activity Test in Fermented Milk

Figure 4 shows the grapefruit extract concentration in the grapefruit extract added fermented milk according to an embodiment of the present invention. In addition, Table 3 shows the acetylcholinesterase-inhibiting activity of the grapefruit extract added fermented milk according to an embodiment of the present invention.

at 0 at 5 dA405 Inhibitory activity (%) Comparative example 0.102 1.242 1.140 0 Example 2-1 (0.1%) 0.102 0.804 0.702 38.4 Example 2-2 (0.5%) 0.102 0.336 0.234 79.4 Example 2-3 (1.0%) 0.102 0.153 0.051 95.5

As shown in Table 3, the comparative example without the grapefruit extract was acetylcholinesterase-inhibitory activity was 0, Example 2-1 containing 0.1% grapefruit extract 38.4%, 0.5% grapefruit extract Example 2-2 containing 79.4%, the fermented milk of Example 2-3 containing 1.0% grapefruit extract exhibits acetylcholinesterase-inhibitory activity of 95.5%, the acetyl of the grapefruit extract added fermented milk according to the present invention It can be seen that the cholinesterase-inhibitory effect is very excellent.

Experimental Example  5: in vivo effects of fermented milk ( in vivo )

In order to determine whether fermented milk added with grapefruit extract showed cognitive improvement (treatment of cognitive dysfunction) in vivo, experimental animals were treated with ICR mice and induced memory loss with scopolamine. (Scop), the general fermented milk feed treatment (Con) and the fermented milk feed treatment (AChE-I, 0.5%) of Example 2-2 was observed the response to the underwater maze experiment (Morris water maze). Underwater maze experiments show that mice damaged by the hippocampus system have obvious obstacles to the learning and memory of spatial relationships required in the Morris underwater maze. In this task, the rat is placed in a cylinder filled with opaque liquid, in which the platform is submerged. The platform is always in a constant place compared to the various visual cues that are located outside the maze (eg objects such as lights, doors, windows around the room). Each trial rat is placed in a cylinder from different positions around the cylinder. Over time, normal rats without damage to the hippocampus system learn the relationship between cues and platform location, and the time it takes to swim and reach the platform is dramatically reduced. In contrast, mice damaged by the hippocampal system are unable to learn the relationship between the cues and find the location of each trial platform, and thus, it is a memory improvement biological experiment that does not shorten the time to reach the platform even after repeated trials.

Figure 5 is a graph showing the memory enhancement effect of the fermented milk added grapefruit extract according to an embodiment of the present invention, the platform arrival time of the general fermented milk feed treatment (Con) in 5 days experiment is excellent as 65.1 seconds, scopolamine Scop, which caused memory loss with (scopolamine), decreased to 88.5 seconds. On the other hand, in the case of fermented milk feeding treatment (AChE-I, 0.5%) of Example 2-2 according to the present invention, the platform reaching time is the shortest as 47.1 seconds, the grapefruit extract added fermented milk of the present invention has excellent memory (cognitive function) improvement effect It could be confirmed that it represents.

Claims (11)

Food composition for preventing and improving cognitive impairment containing grapefruit extract as an active ingredient. According to claim 1, Grapefruit extract is a food composition for preventing and improving cognitive dysfunction that is obtained by hot water extraction of grapefruit. The food composition for preventing and improving cognitive dysfunction according to claim 1, wherein the grapefruit extract is extracted with a solvent selected from one or more alcoholic solvents selected from the group consisting of methanol, ethanol, propanol and butanol. The food composition of claim 1, wherein the grapefruit extract is extracted with a solvent selected from one or more organic solvents selected from the group consisting of acetone, chloroform, methylene chloride, ether, ethyl acetate and hexane. delete delete A health supplement comprising a food composition for preventing and improving cognitive dysfunction of any one of claims 1 to 4. The dietary supplement of claim 7, comprising 0.05 to 5% by weight of the total weight of the food composition for preventing and improving cognitive dysfunction. 8. The dietary supplement according to claim 7, comprising 0.05 to 5% by weight of food composition for preventing and improving cognitive dysfunction and 95 to 99.95% by weight of fermented milk. 10. The dietary supplement of claim 9, wherein the main ingredient of the fermented milk is choline fortified crude oil.
The dietary supplement of claim 10, wherein the choline fortified crude oil contains choline 0.01 to 0.03% by weight relative to the total weight of the crude oil.

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