Food composition comprising hemp seed cake as an active ingredient for enhancing GABA in serum and cerebral cortex and preparation method therefor

Abstract

The present disclosure relates to a food composition comprising hemp seed cake as an active ingredient for enhancing GABA in serum and the cerebral cortex, wherein GABA is enhanced through edestin, an active component contained in the hemp seed cake. According to the present invention, the hemp seed cake extracted independently can provide a large amount of protein and dietary fiber without tolerance, and particularly, through edestin contained in the hemp seed cake, can enhance GABA in serum and the cerebral cortex and induce sleep.

Description

Food composition for enhancing GABA in serum and cerebral cortex containing hemp seed cake as an active ingredient and method for producing the same

[0001] The present invention relates to a food composition for enhancing GABA in serum and cerebral cortex, containing hemp seed cake as an active ingredient, and a method for producing the same, and more particularly, to a food composition for enhancing GABA in serum and cerebral cortex, containing hemp seed cake as an active ingredient, which can provide a large amount of protein and dietary fiber, etc., through hemp seed cake (HSC) extracted by itself, has no resistance, and can enhance GABA in serum and cerebral cortex and induce sleep through the active ingredient Edestin in the hemp seed cake, and a method for producing the same.

[0002]

[0003] One in ten adults suffers from insomnia, which is defined by various factors such as sleep quality, duration, and continuity, and the prevalence varies from 5% to 50% depending on the sleep disorder category.

[0004] Insomnia has been shown to be comorbid with fibromyalgia, peptic ulcer disease, angina, tachycardia, mood disorders, and anxiety disorders. A population-based prospective cohort study of 3,757 Koreans aged 40 to 69 years found an association between prolonged habitual sleep latency and an increased risk of mortality, particularly cancer mortality. Furthermore, subjects in the prospective cohort study with persistent insomnia had a 98% higher mortality rate, higher cardiopulmonary mortality, and higher serum C-reactive protein (CRP) levels.

[0005] Insomnia can have a serious impact on an individual's health and quality of life, and while treatment approaches may vary, there is currently no ideal medication.

[0006] Therefore, while several studies have recently been conducted on the effects of natural products on improving sleep, their effectiveness remains insufficient. However, they have the advantage of having a lower incidence of side effects than conventional medications. Furthermore, many of these natural products, including herbs commonly used to promote sleep, contain flavonoids and terpenoids that act on GABA receptors. GABA receptors are known to play a role in sedation and sleep, anxiety relief, learning and memory, schizophrenia, anesthesia, and neonatal development.

[0007] Furthermore, it is being discovered that cannabinoid receptors (CB1, CB2) play a role in sleep regulation. CCK-positive GABAergic interneuron cell terminals contain CB1 receptors, and propofol, a drug that directly activates GABAa receptors, acts as a competitive inhibitor of FAAH, which degrades the endogenous cannabinoid AEA.

[0008] The most representative medication used for insomnia is benzodiazepine, which is commonly used in various treatments related to anxiety, sleep disorders, seizure disorders, muscle spasms, and depression. Benzodiazepines allosterically modulate GABA and act by binding to the α+ / γ- plane and Cl - They provide therapeutic effects by increasing conduction. However, long-term use of benzodiazepines can lead to tolerance, dependence, withdrawal symptoms, and abuse. While more effective than CBT-1, these treatments are short-term compared to the more long-lasting effects of CBT, and the benefits associated with medication tend to disappear after discontinuation. Side effects, such as cognitive impairment and increased risk of accidents, are more common in older adults, leading to the recommendation against using benzodiazepines for insomnia in older patients.

[0009]

[0010] Meanwhile, hemp seeds are a wonderful natural food that has recently gained popularity. Whole hemp seeds contain 25-35% oil, 20-25% protein, 20-30% carbohydrates, 10-15% insoluble fiber, and vitamins and minerals (phosphorus, potassium, magnesium, sulfur, calcium, iron, and zinc) (Callaway 2004). After removing the hull, the edible portion of the seeds contains an average of 46.7% oil and 35.9% protein.

[0011] Here, it is known that the major protein of hemp seed is edestin, accounting for approximately 65% ​​of the total protein content. When nine types of hemp seed proteins defatted with hexane were subjected to SDS-PAGE, edestin, albumin, and vicilin were separated as representative proteins. The relative density of edestin was 57-76%, while albumin was second at 12-24%.

[0012] Hemp Seed Cake (HSC) is the residue from hemp seeds after the oil has been removed, containing protein and dietary fiber. While considerable research exists on the cardiovascular and cholesterol-lowering properties of hemp seed oil, research on the health benefits of hemp seed cake (HSC), the residue after the oil has been removed, is lacking.

[0013] Although there have been recent studies on the antioxidant, antihypertensive, renal disease control, AChE inhibition, α-glucosidase inhibition, anticancer, antifatigue, and hypercholesterolemia properties of proteins from hemp seed cake (HSC), these are mostly in vitro experiments.

[0014] Furthermore, in vivo studies on hemp seed cake (HSC) have primarily focused on verifying its value as a feed resource for livestock. A study found that mixing hemp seed cake (HSC) into dog food and administering it reduced liver markers, cholesterol, renal function, and biomarkers (creatinine). Furthermore, a study demonstrated anti-anxiety and hypnotic effects in 3-month-old rats.

[0015]

[0016] Accordingly, in the present invention, by extracting hempseed cake (HSC) on its own, it is possible to provide a large amount of protein and dietary fiber, and by conducting animal experiments such as isoflurane-induced sleep tests and GABA and glutamate measurements, it is verified that it can increase GABA in the serum and cerebral cortex without tolerance and induce sleep, thereby confirming that it can be used as a food composition, thereby completing the present invention.

[0017]

[0018] The present invention has been devised to solve the problems of the above-mentioned prior art, and the problem to be solved in the present invention is to provide a food composition for increasing GABA in serum and cerebral cortex, which contains hemp seed cake as an effective ingredient.

[0019] Another purpose is to provide a food composition that can provide a large amount of protein and dietary fiber, etc., through self-extracted hemp seed cake, has no resistance, and, in particular, can increase GABA in the serum and cerebral cortex and induce sleep through the active ingredient Edestin in the hemp seed cake.

[0020]

[0021] In order to achieve the above purpose, a food composition for enhancing GABA in serum and cerebral cortex containing hemp seed cake as an active ingredient according to the present invention is characterized in that GABA is enhanced through the active ingredient Edestin in the hemp seed cake.

[0022] In addition, it is characterized by containing 10 to 50% by weight of the above hemp seed cake as an effective ingredient.

[0023] In addition, the food composition is characterized in that it is in the form of a capsule, tablet, pill, liquid or drink.

[0024] In addition, in the method for producing a food composition for enhancing GABA in serum and cerebral cortex containing hemp seed cake as an active ingredient, the method is characterized by freeze-drying hemp seeds in a vacuum at -40 to -35°C for 48 to 72 hours, then separating the freeze-dried hemp seeds into hemp seed cake and hemp seed oil by extracting them with a low-temperature extractor, and then crushing the separated hemp seed cake into a size of 100 to 1,000 mesh.

[0025]

[0026] According to the present invention, a large amount of protein and dietary fiber can be provided through self-extracted hempseed cake (HSC), and there is no resistance, and in particular, the active ingredient Edestin in the hempseed cake has the effect of increasing GABA in the serum and cerebral cortex and inducing sleep.

[0027]

[0028] Figure 1 is a drawing showing the results of a sleep induction test after a single administration of a hemp seed cake according to the present invention to a mouse.

[0029] Figure 2 is a drawing showing the results of a sleep induction test after administering the hemp seed cake according to the present invention to mice for 15 days.

[0030] Figure 3 is a diagram showing the concentrations of glutamate and GABA in serum and cerebral cortex after a single administration of the hemp seed cake according to the present invention to a mouse.

[0031] Figure 4 is a diagram showing the concentrations of glutamate and GABA in serum and cerebral cortex after administering the hemp seed cake according to the present invention to mice for 15 days.

[0032] Figure 5 is a diagram showing the results of a sleep induction test after administering the active ingredient Edestin to mice for 15 days.

[0033] Figure 6 is a diagram showing the concentrations of glutamate and GABA in serum after administering the active ingredient Edestin to mice for 15 days.

[0034] Figure 7 is a diagram showing the concentration of glutamate and GABA in the cerebral cortex after administering the active ingredient Edestin to mice for 15 days.

[0035] Figures 8 and 9 show GABA and GABA in the cerebral cortex after administering the active ingredient Edestin to mice for 15 days. A This is a diagram showing the receptor content.

[0036]

[0037] Hereinafter, a preferred embodiment of the present invention will be described in detail with reference to the attached drawings.

[0038]

[0039] The present invention relates to a food composition for enhancing GABA in serum and cerebral cortex, containing hemp seed cake as an active ingredient.

[0040] The hemp seed cake according to the present invention is manufactured by freeze-drying hemp seeds in a vacuum at -40 to -35°C for 48 to 72 hours, then extracting the freeze-dried hemp seeds using a low-temperature extractor to separate them into hemp seed cake and hemp seed oil, and then grinding the separated hemp seed cake into a size of 100 to 1,000 mesh.

[0041] More specifically, 100 parts by weight of hemp seeds are freeze-dried in a vacuum in a freeze dryer at -40 to -35°C for 48 to 72 hours, and then the freeze-dried hemp seeds are extracted with a low-temperature oiler to separate them into 60 to 65 parts by weight of hemp seed cake and 35 to 40 parts by weight of hemp seed oil, and the separated hemp seed cakes are crushed 20 to 25 times in 100 to 1,000 mesh for 4 to 6 hours using a 10kW pin crusher.

[0042] The above-mentioned manufactured hemp seed cake (HSC) is the residue from hemp seeds after removing moisture and separating oil, and contains a large amount of protein and dietary fiber.

[0043] In addition, as a natural extract, it is not only non-cytotoxic, but also has no resistance in mouse animal experiments, and it was confirmed that the active ingredient Edestin in the hemp seed cake increases GABA in the serum and cerebral cortex and induces sleep.

[0044]

[0045] The food composition for enhancing GABA in serum and cerebral cortex containing the hemp seed cake of the present invention as an effective ingredient preferably contains 10 to 50 wt% of the extract as an effective ingredient based on the total weight of the composition.

[0046] In order to achieve the minimum effect, it is preferable that the content of the composition be greater than the minimum value mentioned above, and in consideration of the deterioration of the usability due to excessive addition and the applicability to various formulations, it is preferable that the content of the composition be less than the maximum value mentioned above.

[0047]

[0048] In addition, the food composition according to the present invention may further include Gastrodia elata, Atractylodes rhizome, and Poria cocos.

[0049] The above-mentioned Gastrodia elata, Seokchangpo, and Poria cocos provide an anxiety-relieving effect to modern people who have trouble sleeping well due to anxiety and tension, and as a result, their sleep quality is poor and their physical strength is reduced. In the long term, they can provide effects such as physical recovery and memory improvement.

[0050] The above-mentioned Gastrodia elata, Atractylodes japonica, and Poria cocos may each be included in amounts of 15 to 20 parts by weight based on 100 parts by weight of the total food composition.

[0051]

[0052] First, Gastrodia elata is a perennial herb belonging to the Orchidaceae family of the Orchid order of monocotyledons, and its rhizome is used as a medicinal herb. Gastrodia elata is dried to a moisture content of 2-3%, then crushed to a particle size of 300-1,000 mesh before use.

[0053] Gastrodia elata is effective in preventing vascular disease, improving brain health, calming the mind and body, improving insomnia, nourishing the body, preventing aging, improving intestinal health, treating hemiplegia, speech disorders, high blood pressure, epilepsy in children, and epidemic meningitis.

[0054] If the amount of Gastrodia elata included is less than the above range, the effective ingredients of Gastrodia elata cannot be sufficiently provided, and if the amount included exceeds the above range, the unique taste and aroma of Gastrodia elata may become too strong, which may actually lower the overall taste.

[0055]

[0056] Aster is a perennial herb of the Araceae family in the Araceae order of monocotyledons. Its rhizome is used. The aster is dried to a moisture content of 2-3%, then crushed to a particle size of 300-1,000 mesh.

[0057] The rhizome of this aloe vera plant contains essential oil, tannin, vitamin C, starch, and palmitic acid, and the essential oil contains eugenol, sesquiterpene, and asarone, which clear the mind, promote blood circulation, and have calming, analgesic, antispasmodic, and circulatory system effects, strengthen the immune system, and antioxidant effects. In particular, the essential oil components such as palmitic acid, asarone, and phenol contained in the rhizome of the aloe vera plant have the effect of improving the fragrance.

[0058] If the amount of Seokchangpo is less than the above range, the effective ingredients of Seokchangpo cannot be sufficiently provided, and if the amount is exceeded, the unique taste and smell of Seokchangpo may become too strong, which may actually lower the overall taste.

[0059]

[0060] The above-mentioned Poria cocos is a dried fungus belonging to the Polyporaceae family, containing the pine root in the center of the tuber. It contains 7.5% of β-pacyman and triterpenoid compounds such as pachymic acid, ebricoic acid, tumulosic acid, and pinicolic acid, so it is effective for symptoms such as nervous stability, concentration, and insomnia.

[0061] These aphids are dried to a moisture content of 5-7%, then crushed to a particle size of 300-1,000 mesh before use.

[0062] If the amount of Fuxin is less than the above range, Fuxin's excellent effects of nerve stability and concentration will be minimal, and if it is included beyond the above range, Fuxin's unique taste and aroma will become too strong, which may actually lower the overall taste.

[0063]

[0064] Additionally, the food composition according to the present invention may further comprise one or two of vitamin B and astaxanthin.

[0065] The above vitamin B and astaxanthin may be included as auxiliary ingredients to mask the off-flavor and taste of a food composition for enhancing GABA in brain tissue containing hemp seed cake as an active ingredient.

[0066] The above vitamin B and astaxanthin may each be included in an amount of 0.01 to 10 parts by weight based on 100 parts by weight of the entire food composition, and the content range may be adjusted according to requirements such as the stability and content of the food composition for promoting GABA in brain tissue containing hemp seed cake as an active ingredient, and the ease of formulation of the composition.

[0067] The above food composition may include a food additive that is food-wise acceptable.

[0068] The above food additives refer to components that can be added to food for preservation purposes, and can be appropriately selected and used by those skilled in the art as they are added in the manufacture of health functional foods of each formulation.

[0069] Examples of food additives include, but are not limited to, various nutrients, vitamins, minerals (electrolytes), colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages.

[0070] Additionally, the food composition may include a health functional food.

[0071] In this specification, "health functional food" refers to a food group that has been given added value by using physical, biochemical, or bioengineering techniques to enable the function of the food to function and express for a specific purpose, or a food that has been designed and processed so that the body's body's regulatory functions, such as regulating the biological defense rhythm and preventing and recovering from diseases, are fully expressed. Compared to general foods, it has an active health maintenance or promotion effect, and health supplement food refers to a food for the purpose of health supplementation. In some cases, the terms functional food, health food, and health supplement food are used interchangeably.

[0072] The above food may contain food additives that are food-scientifically acceptable, and may further contain suitable carriers, excipients, and diluents commonly used in the manufacture of health functional foods.

[0073]

[0074] The food composition of the present invention may include, in addition to the active ingredient, a sweetener, a flavoring agent, a physiologically active ingredient, and minerals.

[0075] Sweeteners can be used in amounts that impart an appropriate sweetness to the food and can be natural or synthetic. Natural sweeteners are preferred, including corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

[0076] Flavoring agents can be used to enhance taste or aroma, and both natural and synthetic flavors can be used. Natural flavoring agents are preferred. When using natural flavoring agents, they can also serve the purpose of enhancing nutrition in addition to flavor. Natural flavoring agents can be obtained from apples, lemons, tangerines, grapes, strawberries, and peaches, or from green tea leaves, Polygonum multiflorum, bamboo leaves, cinnamon, chrysanthemum leaves, and jasmine. Ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo biloba can also be used. Natural flavoring agents can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents can be used, such as esters, alcohols, aldehydes, and terpenes.

[0077] Physiologically active substances that can be used include catechins such as catechin, epicatechin, gallogatechin, and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, and riboflavin.

[0078] Minerals that can be used include calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, and zinc.

[0079] In addition, the food composition of the present invention may include, in addition to the sweetener, etc., a preservative, an emulsifier, an acidulant, a thickener, etc., as needed.

[0080] Preservatives, emulsifiers, and the like are preferably added in trace amounts that can achieve their intended purpose. "Trace amounts," expressed numerically, refer to a range of 0.0005 to 0.5 wt% based on the total weight of the food composition.

[0081] Preservatives that can be used in the present invention include sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, and EDTA (ethylenediaminetetraacetic acid).

[0082] Emulsifiers include acacia gum, carboxymethyl cellulose, xanthan gum, and pectin.

[0083] Acidulants include lactic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. In addition to enhancing flavor, these acidulants may be added to food compositions to maintain an appropriate acidity level for the purpose of inhibiting microbial growth.

[0084] Thickening agents include suspending agents, sedimentation agents, gelling agents, and bulking agents.

[0085]

[0086] In addition, the health functional food of the present invention can be manufactured by mixing other appropriate auxiliary ingredients and known additives that can be included in foods according to the selection of a person skilled in the art. Examples of foods to which the health functional food of the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and the food extract of the present invention can be manufactured by adding it to juice, tea, jelly, and juice manufactured using the food extract of the present invention as a main ingredient.

[0087]

[0088] The above food composition may be in the form of a capsule, tablet, pill, liquid or drink.

[0089] If the food formulation is a beverage, it may contain additional ingredients such as various flavorings or natural carbohydrates, just like regular beverages.

[0090] The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the above-mentioned natural carbohydrate is 0.01 to 0.04 parts by weight, preferably 0.02 to 0.03 parts by weight, per 100 parts by weight of the composition of the present invention.

[0091] In addition to the above, the food formulation may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food formulation may contain fruit pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination. The proportion of these additives is not particularly critical, but is selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0092]

[0093] The present invention is further described in detail below through examples. However, it should be clear to those skilled in the art that the following examples are intended only to illustrate the present invention and do not limit its scope. In other words, simple modifications and variations of the present invention can be readily implemented by those skilled in the art, and all such modifications and variations are considered to fall within the scope of the present invention.

[0094]

[0095] Example: Hemp seed cake (HSC) according to the present invention

[0096] 10 kg of hemp seeds were freeze-dried in a vacuum in a freeze dryer at -40°C for 72 hours, and then the freeze-dried hemp seeds were extracted using a low-temperature extractor to separate them into 6.1 kg of hemp seed cake and 3.9 kg of hemp seed oil, respectively. The separated hemp seed cake was then crushed 20 times at 300 mesh using a 10 kW pin crusher for 5 hours to produce the product.

[0097]

[0098] Experiment 1: Analysis of major proteins in hemp seed cake (example, HSC) according to the present invention using Nano-LC-MS / MS

[0099] Nano-LC-MS / MS analysis was performed to identify the main active ingredient of the hemp seed cake (Example, HSC) according to the present invention. The protein concentration of the hemp seed cake (Example, HSC) was measured to be 3.4 mg / ml using the Bradford method, and protein separation and purification were performed.

[0100] Finally, 1 mg of HPC (Hempseed protein concentrate) was obtained, and the results obtained through LC-MS / MS were compared with previously reported hempseed proteins (Aiello, Lammi et al. 2017). Nine proteins were identified and their relative amounts were calculated. The results are shown in Table 1.

[0101] As can be seen in Table 1, the sum of Edestin1 and Edestin2 showed the highest content ratio at 31.25%.

[0102] Proteinrelative abundance(%)Polyketide synthase 4 (PKS4)16.19Edestin216.03Edestin115.224-Coumarate : CoA ligase14.42THCA synthase13.94photosystem I P700 chlorophyll A apoprotein9.62naringenin chalcone synthase8.81oxidoreductase chain 53.37Protein CcsA2.40

[0103]

[0104] Experiment 2: Animal Experiment

[0105] 1) Animal model

[0106] All animal studies followed the guidelines of the Chungbuk National University Institutional Animal Care and Use Committee (IACUC) and adhered to the Chungbuk National University Animal Care and Use Committee. Eight-week-old male B6 mice (weighing 30 g) were purchased and acclimated to the facility for one week prior to experimental use. Mice were fed standard diet and maintained under controlled environmental conditions (12-h light / dark cycle, 23±2°C, and 55±10% humidity).

[0107] The hemp seed cake of the present invention (Example, HSC), chlorpromazine (positive control group) and edestin (Edestin) were administered to 5 animals per group, and the control group and the positive control group were set as 4 mg / kg and 8 mg / kg chlorpromazine groups.

[0108]

[0109] 2) Sleep induction test

[0110] The hemp seed cake (example, HSC) administration group of the present invention was administered orally once a day at 900 mg / kg, considering the body weight of 30 g of the mouse, by mixing 27 mg of the hemp seed cake (example, HSC) of the present invention in 0.3 ml of distilled water (5 mice each of 50, 100, and 150 mg, 405 mg of the hemp seed cake (example, HSC) of the present invention in 4.5 ml of distilled water, and fed 0.1 ml / 0.2 ml / 0.3 ml to a total of 15 mice). The short-term (n=5 per group) group was administered the hemp seed cake of the present invention (example, HSC) only once 1 hour before the sleep test, while the long-term group (n=5 per group) was administered the hemp seed cake of the present invention (example, Hemp Seed Cake, HSC) daily for 15 days and underwent a sleep test on the last day.

[0111] After each oral administration, anesthesia was induced by placing the mice in an anesthesia box filled with 4% isoflurane, prepared by mixing it with air using a vaporizer, and leaving them in that condition for 2 minutes. The mice were then removed from the anesthesia box, placed on their backs, and the time to recovery from loss of righting reflex (LR) was measured. Loss of R was defined as the inability of the mouse to maintain its position on all four paws, and recovery of R was defined as the moment when the mouse was able to maintain its position on all four paws for the first time. The results are presented in Figures 1 and 2.

[0112] As shown in FIGS. 1 and 2, the experimental results showed that the time required to recover from loss of righting reflex increased in a dose-dependent manner in the hemp seed cake (example, HSC) group of the present invention compared to the control group, demonstrating that the hemp seed cake (example, HSC) of the present invention is effective in inducing sleep.

[0113] In particular, as shown in Fig. 2, it was found that mice administered 900 mg / kg of the hemp seed cake of the present invention (Example, HSC) daily for 15 days showed results similar to those of the positive control group of 4 mg / kg.

[0114] In addition, when comparing Figs. 1 and 2 with respect to resistance, it was observed that the positive control group developed resistance after 15 days of administration and a decrease in effectiveness was observed, but when the hemp seed cake of the present invention (Example, HSC) was administered for 15 days, it was confirmed that the decrease in effectiveness due to resistance was not significant.

[0115]

[0116] 3) Experiment to measure glutamate and GABA concentrations in serum and cerebral cortex

[0117] Homogenized serum and brain tissue samples were analyzed using a Shimadzu Prominence LC-20AD HPLC system equipped with an electrochemical detector Nanospace 3005 (OSAKA SODA, Tokyo, Japan) using a Unison UK-C18 column (IMTAKT, Kyoto, 2.0 mm × 50 mm). The graphite working electrode of the electrochemical detector was set at +850 mV with respect to the Ag / AgCl reference electrode.

[0118] The mobile phase consisted of 50 mM phosphoric acid, 50 mM citric acid, 0.1 mM EDTA (pH 3.5), and 5% acetonitrile at a flow rate of 0.3 mL / min. Twenty microliters of sample was added to 2 μL of a reactive solution prepared with o-phaldialdehyde (37 mM) in 0.1 M borate buffer (pH 10.4) containing 250 μL ethanol and 50 mM sodium sulfite. The sample (10 μL) was then injected into the HPLC system. The amount of neurotransmitter in the dialysate was calculated using Labsolutions software (Simadzu, Kyoto, Japan). The results are shown in Figures 3 and 4.

[0119] As shown in Figures 3 and 4, glutamate was not significant in serum and brain tissue, but GABA increased dose-dependently in serum after 15 days of administration and increased compared to the control group, and GABA increased dose-dependently in cerebral cortex tissue after single administration and 15 days of administration, and the cumulative effect increased compared to the positive control group, and no tolerance was observed.

[0120] In particular, the increase in GABA was greater in the cerebral cortex than in the serum in the positive control group and the hemp seed cake (example, HSC) group of the present invention, and it was confirmed that the positive control group and the hemp seed cake (example, HSC) of the present invention can increase GABA in the cerebral cortex tissue. The hemp seed cake (example, HSC) of the present invention increased GABA in the cerebral cortex tissue to a level similar to that of the positive control group, and the 900 mg / kg group of the hemp seed cake (example, HSC) of the present invention showed a greater increase in GABA than the 4 mg / kg group of the positive control group.

[0121]

[0122] 4) Delayed recovery of righting reflex by edestin

[0123] Based on the administration of 150 mg / kg, 4.5 mg of edestin was mixed in 0.3 ml of distilled water and administered orally once a day, considering the body weight of 30 g of mice (15 mice in total, 5 mice each of 50, 100, and 150 mg, 4.5 mg of edestin mixed in 4.5 ml of distilled water, administered 0.1 ml / 0.2 ml / 0.3 ml). The righting reflex recovery time was measured in the same manner as before. The results are shown in Fig. 5.

[0124] As shown in Fig. 5, mice orally administered Edestin for 15 days showed a significant dose-dependent increase in the righting reflex recovery time from anesthesia with 4% isoflurane, and when 150 mg / kg of Edestin was administered, the righting reflex recovery time was similar to that of 900 mg / kg of the hemp seed cake of the present invention (Example, HSC). These results indicate that the main active ingredient for inducing sleep in the hemp seed cake of the present invention (Example, HSC) is Edestin.

[0125]

[0126] 5) Experiment to measure glutamate and GABA concentrations in serum and cerebral cortex using edestin

[0127] To investigate the mechanism of sleep-inducing effects of edestin, serum and cerebral cortex tissue were collected from mice after the experiment, and glutamate and GABA concentrations were measured using ELISA. The results are shown in Figures 6 and 7.

[0128] As shown in Fig. 6, serum glutamate decreased in the group administered Edestin compared to the control group, and the decrease was significant in a dose-dependent manner. Serum GABA increased in a dose-dependent manner in the group administered Edestin compared to the control group, confirming that Edestin can decrease serum glutamate and increase GABA.

[0129] As shown in Figure 7, there was no significant difference in glutamate in the cerebral cortex tissue in the group administered Edestin compared to the control group, and GABA significantly increased in a dose-dependent manner in the group administered Edestin compared to the control group, confirming that Edestin can increase GABA in the cerebral cortex.

[0130] The increase in GABA was greater in the cerebral cortex than in the serum in the group administered edestin. In addition, the cerebral cortical GABA concentration of mice administered 150 mg / kg of edestin was slightly lower than that of mice administered 900 mg / kg of the hemp seed cake of the present invention (example, HSC), but was almost at a similar level. These results indicate that the main active ingredient for inducing sleep in the hemp seed cake of the present invention (example, HSC) is edestin.

[0131]

[0132] 6) GABA and GABA in the cerebral cortex using Western blot A receptor testing

[0133] To investigate the mechanism of the effect of edestin on sleep induction, Western blot analysis was performed to determine the levels of GABA and GABA in the cerebral cortex of mice administered edestin for 15 days. A The receptor content was investigated. The results are shown in Fig. 8.

[0134] As shown in Figure 8, GABA and GABA by Edestin A It was confirmed that the receptor expression increased in a dose-dependent manner.

[0135]

[0136] 7) GABA and GABA in the cerebral cortex using immunohistochemistryA receptor testing

[0137] To investigate the mechanism of the effect of edestin on sleep induction, immunohistochemical examination was performed on GABA and GABA in the cerebral cortex of mice administered edestin for 15 days. A The receptor was stained. The results are shown in Figure 9.

[0138] As shown in Figure 9, GABA and GABA by Edestin A It was confirmed that the receptor expression increased in a dose-dependent manner.

[0139]

[0140] According to the present invention, a large amount of protein and dietary fiber can be provided through self-extracted hempseed cake (HSC), and there is no resistance, and in particular, the active ingredient Edestin in the hempseed cake has the effect of increasing GABA in the serum and cerebral cortex and inducing sleep.

Claims

1. A food composition for enhancing GABA in serum and cerebral cortex, containing hemp seed cake as an active ingredient.

2. In paragraph 1, A food composition for increasing GABA in serum and cerebral cortex, characterized in that GABA is increased through the active ingredient Edestin in the above hemp seed cake.

3. In either paragraph 1 or paragraph 2, A food composition for enhancing GABA in serum and cerebral cortex, containing 10 to 50 wt% of hemp seed cake as an active ingredient.

4. In either paragraph 1 or paragraph 2, The above food composition, A food composition for enhancing GABA in serum and cerebral cortex, comprising hemp seed cake as an active ingredient, characterized in that it is in the form of a capsule, tablet, pill, liquid or drink.

5. A method for producing a food composition for enhancing GABA in serum and cerebral cortex containing hemp seed cake as an effective ingredient, A method for producing a food composition for enhancing GABA in serum and cerebral cortex, comprising: freeze-drying hemp seeds in a vacuum at -40 to -35°C for 48 to 72 hours, extracting the freeze-dried hemp seeds with a low-temperature extractor to separate them into hemp seed cake and hemp seed oil, and grinding the separated hemp seed cake into a size of 100 to 1,000 mesh.

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