KR101219627B1 - Composition containing extracts from rice bran oil for prophylaxis or treatment of cerebrovascular diseases and method of production thereof - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating cerebral disease using rice bran oil extract as an active ingredient, and more specifically, as a composition for preventing or treating brain disease or thermosetting neuroprotective cells containing thermocompressed rice bran oil extract extract as an active ingredient, The extract of rice bran oil is not only excellent in inhibiting neuronal cell death caused by cerebral ischemia and inhibiting memory damage but also effectively used without side effects to prevent and treat brain diseases caused by necrosis of nerve cells due to harmless to human body. Could be.

Description

Composition containing extracts from rice bran oil for prophylaxis or treatment of cerebrovascular diseases and method of production according to rice bran oil extract

The present invention relates to a pharmaceutical composition for preventing or treating cerebral diseases using rice bran oil extract as an active ingredient, and more particularly, to a composition for preventing or treating brain disease or thermosetting neuroprotective cells containing thermocompressed rice bran oil extract as an active ingredient. will be.

In today's developed and some developing countries, disease structure is altered by industrialization, urbanization, lower mortality rates and increased life expectancy. In terms of the disease aspect, the incidence of infectious diseases, which have been a traditional health problem in the past, has decreased, and the proportion of non-communicable and chronic degenerative diseases has been increasing. It is a trend.

On the other hand, as the average GNP of $ 10,000 was entered, people became more concerned about their health, and the average life expectancy increased rapidly, so that the average life expectancy for women is 80 years or older. Increasing life expectancy is important for chronic degenerative brain disease, which has not been important in the past, and studies are being actively conducted to prevent and treat it.

In the case of transient cerebral ischemia in the cerebrum, the supply of oxygen and glucose is blocked, resulting in a decrease in ATP and edema in neurons, which eventually leads to extensive brain damage. Neuronal death occurs after a significant period of time after cerebral ischemia, which is called delayed neuronal death. Delayed neuronal death was observed in a transient forebrain ischemic model using Mongolian gerbil, and neuronal cell death was observed in the CA1 region of the hippocampus (hippcampus) 4 days after induction of cerebral ischemia for 5 minutes. (Kirino T, Sano K. Acta Neuropathol., 62, pp 201-208, 1984; Kirino T. Brain Res., 239, pp 57-69, 1982).

The causes of neuronal cell death due to cerebral ischemia are the excitatory neuronal cell death (Excitotoxicity, Kang TC, Hwang IK, Park SK, An). SJ, Yoon DK, Moon SM, Lee YB, Sohn HS, Cho SS, Won MH.J Neurocytol. 30 (2001) 945-955) and DNA damage caused by reactive oxygen radicals occurs early and eventually Oxidative damage (Won MH, Kang TC, Jeon GS, Lee JC, Kim DY, Choi EM, Lee KH, Choi CD, Chung MH, Cho SS. Brain Res. 836 (1999) 70-78; Sun AY, Chen YM. J Biomed Sci. 5 (1998) 401-414, Flowers F, Zimmerman JJ. New Horiz. 6 (1998) 169-180). However, these two mechanisms of injury are thought to be induced simultaneously to aggravate neuronal damage (Won MH, Kang T, Park S, Jeon G, Kim Y, Seo JH, Choi E, Chung M, Cho SS. Neurosci Lett 301 (2001) 139-142).

Several test methods have been proposed that resemble the mechanisms of brain damage caused by these various causes, one of which is cerebral ischemia-reperfusion. The mechanism of brain cell damage caused by this method can be summarized in two ways. One is the “excitotoxicity” theory that excessive calcium ions enter neurons and neurons die as the glutamate is excessively increased after cerebrovascular perfusion and the calcium ion channels open (Hagberg, 2000). H. et al., Ment Retard Dev Disabil Res Rev. 8 (1): 30-38 (2002)). Another hypothesis is the "oxidative stress theory," which is attributed to the breakdown of the intracellular enzyme lineage by free radicals induced during reperfusion (Chan. PH., J Cereb Blood Flow Metab. 21 (1): 2-14 ( 2001). Studies have shown that neuronal cell death by excitatory cytotoxicity and oxygen radicals does not occur separately but in parallel with each other, and they act directly on neuronal death (Won, MH. Et al., Brain). Res. 836: 70-78 (1999). Recently, attention is focused on substances such as inflammatory mediators and cytokines which are mainly induced around 4 days of cell death. These cytokines and inflammatory mediators are known to increase cell death by secreting from microglia or from dying cells (Basu, A. et al., J Neurosci. 22: 6071-6082). (2002)).

Based on these mechanisms, a lot of research has been conducted to search for substances that effectively inhibit neuronal cell death in cerebral ischemia or to preclinically test them. Tissue plasminogen activator is a commercially available drug approved by the FDA as a treatment for cerebral ischemia. Tissue plasminogen activator is originally used as a thrombolytic agent, but when used within 3 hours of cerebral ischemia, it has been found to be effective in preventing neuronal cell death and is used as a treatment for cerebral ischemia. However, tissue plasmidogen activators have been reported to have side effects of dissolving blood clots caused by vascular occlusion and causing bleeding. MK-801, a calcium channel blocker to effectively inhibit early calcium influx, has been clinically tested, but the drug has been discarded due to its side effects.

On the other hand, in Korea, many natural substances are marketed as health foods that are effective in preventing stroke, but most of them are not scientifically verified, and they are often social problems because they cause health food abuse.

Therefore, there is an urgent need for an effort to objectively verify the natural resources that can be eaten as foods and to develop natural materials having the effect of treating and preventing brain diseases.

Accordingly, the present inventors confirmed that rice bran oil has an activity capable of significantly inhibiting cerebral ischemic neuronal cell death while continuing research on the development of a natural substance having a therapeutic and preventive effect on brain disease, and completed the present invention based thereon. It was.

An object of the present invention is to provide a natural food extract that is harmless to the human body and can prevent nerve cell damage caused by cerebral ischemia.

It is another object of the present invention to provide a rice bran extract having a brain disease prevention and treatment effect.

In another aspect, an object of the present invention is to provide a composition for preventing or treating brain disease or for protecting nerve cells comprising the rice bran oil extract as an active ingredient.

In order to achieve the above object, the present invention provides a rice bran oil extract for nerve cell protection prepared by shaking a mixture of rice bran oil and an organic solvent.

In another aspect, the present invention provides a composition for preventing and treating cerebral disease or protecting the nerve cells of the brain comprising the rice bran oil extract as an active ingredient and a method for preparing the same.

Hereinafter, the present invention will be described in detail.

The composition of the present invention is characterized in that it contains rice bran oil extract, and more specifically, rice bran oil extract, which is used for preventing and treating brain diseases caused by neuronal cell death, and for preventing or treating brain diseases containing the same or for protecting brain cells. Characterized by providing a composition.

In addition, the method for producing a composition of the present invention comprises the steps of obtaining rice bran oil by thermal compression; And extracting the obtained rice bran oil with an organic solvent.

As used herein, the term "prevention" means that it has not been diagnosed as having a disease or condition but inhibits the occurrence of the disease or condition in an animal that tends to be prone to such disease or condition. As used herein, the term “treatment” refers to (a) inhibiting the development of a disease or condition; (b) alleviation of the disease or condition; And (c) elimination of the disease or condition.

The present invention provides an active ingredient, a composition for preventing or treating brain disease or protecting nerve cells, containing rice bran oil extract.

Rice bran oil is a high quality oil extracted and purified from rice germ and rice bran, and it contains squalene to promote healthy skin maintenance and metabolism, and contains tocotrienol and tocopherol to have antioxidant effect, chembesterol, It contains cytosterol, stigmasterol and oryzanol to promote blood circulation, and contains other vitamins to play an important role in skin moisturization and skin cell respiration.

In the present invention, the rice bran oil extract is a thermocompressed rice bran oil extract, and more specifically, the thermocompressed rice bran oil extract is a step of obtaining a rice bran oil by thermocompressing rice bran; Extracting the obtained rice bran oil with an organic solvent; will be obtained.

Brain disease to which the composition of the present invention can be applied is one or more selected from cerebral infarction, cerebral edema, cerebral ischemia, vascular dementia and memory damage, but is not limited thereto. More specifically, neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease and amyotrophic lateral sclerosis; Diseases caused by damage to nerve cells following ischemia or reperfusion, such as stroke (especially ischemic stroke); And it can be used for the prevention or treatment of schizophrenia, depression, mood swings and mental disorders such as mental post-traumatic stress disorder.

The present invention

1) thermocompressing rice bran to obtain rice bran oil;

2) extracting the obtained rice bran oil with an organic solvent; And

3) drying the extracted rice bran oil;

It provides a method for producing a composition for preventing or treating brain disease or brain nerve cell protection prepared to include.

In the present invention, the thermocompression of step 1) is characterized in that the compression at 50 to 70 Mpa pressure at a temperature of 70 to 80 ℃.

In the present invention, the extraction of step 2) is characterized in that the extraction after shaking the rice bran oil and the organic solvent in a 1: 3 to 5 weight ratio, the temperature of 30 to 50 ℃.

The organic solvent may be alcohol having 1 to 5 carbon atoms, hexane, chloroform or ethyl acetate, and in one embodiment of the present invention, a mixture of rice bran oil and hexane was prepared by shaking extract, the rice bran oil extract is cerebral ischemia It was confirmed that it can significantly prevent neuronal damage caused by.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of a powder, granules, tablets, capsules, and may further include a dispersant or stabilizer.

The composition of the present invention can be developed into a food composition. Ingredients commonly added in the manufacture of food products include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when prepared with a drink, in addition to staniocalcin 2 of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, and the like may be further included. Given the ease of access to food, the food of the present invention is very useful for the treatment or prevention of neurological diseases, and for improving cognitive function.

In the present invention, rice bran oil extract is not only excellent in inhibiting neuronal cell death caused by cerebral ischemia and suppressing memory damage, but also harmless to the human body, and is used for preventing and treating brain disease caused by necrosis of nerve cells. It can be used effectively.

1 is a photograph showing the effect of inhibiting cerebral infarction by ligation / reperfusion of rice bran oil (RBO) extract according to the present invention,
Figure 2 is a graph showing the effect of inhibiting cerebral infarction of rice bran oil (RBO) extract according to the present invention,
Figure 3 shows the inhibitory effect of behavioral abnormalities according to cerebral infarction of rice bran oil (RBO) extract according to the present invention,
(A: loss of muscle coordination, B: behavioral disorder due to nerve injury)
4 is a result of examining the volume change of infarction according to pre-infarction treatment of rice bran oil (RBO) extract according to the present invention,
5 is a result of investigating behavioral changes according to pre-infarction treatment of rice bran oil (RBO) extract according to the present invention,
(A: loss of muscle coordination, B: behavioral disorder due to nerve injury)
Figure 6 is a result of examining the volume change of infarction following treatment with cerebral bran oil (RBO) extract according to the present invention,
7 is a result of investigating the behavioral change according to post-infarction treatment of rice bran oil (RBO) extract according to the present invention,
(A: loss of muscle coordination, B: behavioral disorder due to nerve injury)
8 is a graph showing the effect of inhibiting memory damage by Aβ (25-35) of rice bran oil (RBO) extract according to the present invention,
9 is a graph confirming the neuronal cell death inhibitory effect of rice bran oil (RBO) extract according to the present invention by MTT colorimetric assay.
(A: Aβ (25-35) -induced neuronal death, B: glutamate-induced neuronal death, C: hypoxia-induced neuronal death)

The present invention will be described in more detail with reference to the following examples. However, the following examples are provided to aid understanding of the present invention, and the scope of the present invention is not limited by these examples in any sense.

Hereinafter, the technical and scientific terms used herein will be understood by those skilled in the art without departing from the scope of the present invention. Descriptions of known functions and configurations that may be unnecessarily blurred are omitted.

Preparation Example Preparation of Rice Bran Oil (RBO) Extract

Rice cultivated in Cheongwon-gun, Chungbuk, Korea, used 18 kg of fresh rice bran sample from Cheongwon Life Rice (Brown Rice) at the Ochang Rice Processing Center (RPC).

18 kg of rice bran samples were milked. First, it was put into the cylinder of the milking mechanism, and when the internal temperature reached 75 ° C., the pressure of the milking machine was constantly increased. After the pressure reached 60 Mpa, it was pressed for 20 minutes and milked to obtain 1,350 ml of rice bran oil. After the milking, the residual fat of the pressed cake was measured according to the AOAC method, and the oil extraction rate calculated as the difference between the total fat and the residual fat was 35%, and the oil contained in the rice bran was 14-17%. Yield of rice bran oil by the law was about 5-8%.

The milking device was milked using a National Hanaro Milking Machine (National ENG), the milking mechanism is made of stainless steel, and the cylinder and the bottom plate are connected by screws. The inner circumference of the cylinder was 27.8 mm, and 50 discs with a diameter of 2 mm were drilled in the disc supporting the sample for oil leakage. Milking was performed by adding two 60 mesh nets to prevent solids from spilling.

After mixing n-Hexane 3.6 L with 1.35 L of the obtained rice bran oil, the extract obtained by shaking extraction at 37 ° C. for 12-24 hours was evaporated to dry all n-Hexane using a rotary evaporator (Rice Bran). Oil) extract was obtained. 1,260 ml of rice bran oil extract was obtained from 18 kg of rice bran, and the obtained rice bran oil was stored at 4 ° C in a shaded state.

Experimental Example 1 Measurement of Inhibitory Effect of Thermocompressed Rice Bran Oil Extract on Cerebral Infarction After Ischemia / Reperfusion by the Middle Cerebral Artery Occlusion (MCAO) Method

300 ± 5 g of Sprague-Dawley (SD) rats were inhaled with 1-2% isoflurane (N2O: O2 = 4: 1), and then surgically operated on the right carotid artery Exposed. The groin artery branching from the external carotid artery to the central part and the thyroid gland branching from the external carotid artery to the internal carotid artery were ligated by electrophoresis. Arrange the area well until a Y-shaped branch is found above the internal carotid artery, and then the head of the external carotid artery is threaded to block blood flow and to prevent blood flow between the total carotid and internal carotid arteries. The bottom was clipped. After inserting a probe through a small hole in the carotid artery, the probe and the lower part of the external carotid artery were lightly tied to the thread just enough to prevent bleeding, and then the clip that blocked the blood flow was removed. After cutting between the hole where the probe was inserted and the tied portion of the external carotid artery, the probe inserted into the external carotid artery was carefully inserted into the internal carotid artery. The probe was inserted into the inner vessel of the blood vessel branching Y-shaped above the internal carotid artery. The probe was inserted only 20 mm from the total carotid artery branch. The rectal temperature was measured immediately after surgery, and the body temperature was maintained at 37 ± 0.5 ° C. using a heating pad for 6 hours. After 2 hours of occlusion, the probe was removed and reperfused. After 24 hours, the rats were dislocated and the brains were separated, cut into 2 mm thickness using a brain matrix, and stained with 2% TTC (2,3,5-triphenyltetrazolium chloride) solution for 30 minutes at 37 ° C. After that, a digital camera was used to measure infarct volume (mm ³ ) using an image analysis system.

The behavioral indicators of nerve injury were 1) the degree of motor coordination by rota rod test and 2) the neurological score by Menzies method.

As a result, as can be seen in Figure 1, the left cerebral tissue when the brain ischemic state in which nothing is processed, it can be seen that the cerebral infarction appeared white in the left hemisphere, compared to the second to fourth lines of the brain from the left 0.5, 1, 3 ml / kg of the extract was administered orally twice an hour before arterial ligation and 1 hour after ligation. A dose of 1 and 3 ml / kg of cerebral infarction due to cerebral ischemia caused by carotid artery ligation It was confirmed that significantly inhibited in.

In addition, as seen in FIG. 2 in the infarct volume, the control group (vehicle group) showed an infarct volume of 308.8 mm 3, Rice Bran Oil Extract The infarct volume was reduced to 227.2 ㎣ in the 0.5 ml / kg group, 195.0 ㎣ in the 1 ml / kg group, 196.1 3 in the 3 ml / kg group, and 237.8 5 in the 5 ml / kg group and 243.8 ㎣ in the 10 ml / kg group. It was confirmed that it was confirmed that the most significantly inhibited infarction in the administration group administered doses of 1 and 3 ml / kg.

In addition, as a result of confirming the inhibitory effect of the rice bran oil extract on the loss of muscle cooperative coordination due to cerebral infarction, as shown in Figure 3, the cooperative exercise disturbance by cerebral infarction most remarkably in the administration group administered doses of 1 and 3 ㎖ / kg It was confirmed that the inhibition, and the inhibitory effect of rice bran oil on behavioral abnormalities caused by neurological damage, it was confirmed that the most severely inhibited behavioral disorder in the administration group administered 1 and 3 ㎖ / kg dose.

From the above results, the volume and behavioral changes of infarct according to the administration of rice bran extract before and after cerebral infarction were selected by selecting 1 and 3 ml / kg of the most effective doses of rice bran extract.

To investigate the volume and behavioral changes of infarction before cerebral infarction, 1 or 3 ml / kg of rice bran oil was administered orally once a week for one week, and the brain was examined 24 hours after surgery and carotid artery ligation 30 minutes after the last dose. Extraction was performed to measure the infarct volume.

As a result, as shown in Figure 4, the control group (vehicle group) showed an infarct volume of 359.0 9.0, Rice Bran Oil Extract 1 ml / kg administration group was confirmed that the infarct volume was reduced to 249.1 ㎣, 3 ml / kg administration group was 272.1 ,, and as shown in Figure 5, the inhibitory effect of the rice bran oil on loss of muscle movement coordination caused by cerebral infarction However, the inhibitory effect of rice bran oil on behavioral abnormalities caused by nerve injury was confirmed.

In addition, in order to investigate the volume and behavioral changes of infarction after cerebral infarction, 1 ml of rice bran extract and 3 ml / kg dose were administered 1 hour after surgery and carotid artery ligation, and after oral administration once daily, thereafter, along with behavioral test on day 7 The brain was harvested and the infarct was measured.

As a result, as can be seen in Figure 6, the control group (vehicle group) showed a growth volume of 302.1 ㎣, Rice Bran Oil Extract 1 ml / kg administration group was confirmed that the infarct volume was reduced to 193.9 ㎣, 3 ㎖ / kg administration group 156.3 ㎣, as shown in Figure 7A, the inhibitory effect of rice bran oil extract on loss of muscle movement coordination by cerebral infarction As a result, the control group (vehicle group) did not change the time to endure in the rota rod, but the rice bran extract administration group was gradually recovered to confirm that the time is significantly longer. In addition, as a result of confirming the behavior change on the 6th day after the operation, as shown in B of FIG.

From the above results, it was confirmed that the rice bran oil extract significantly reduced the infarct volume of rat brain tissue caused by infarction by ischemia / reperfusion, and significantly improved coordination disorder and behavioral abnormality according to cerebral infarction.

[ Experimental Example  2] Manual Evasion Test Step through method Measurement of memory and learning by

Passive Avoidance Test is one of the most common and excellent methods used in rodents to measure the effects of drugs on memory and learning. Step through apparatus (Gemini II avoidance system, Sandiego Instrument) is a shuttle box divided into two compartments (25 (W) × 21 (D) × 19 (H) cm) It is connected by a photo cell-guarded gate, and the bottom is made up of a grid. One side was illuminated to create a bright environment, the other through the door to a dark environment, and electrical stimulation was applied through a grid of dark boxes.

ICR mice were used as animals, and after the purchase, the rice bran oil extract (0.5, 1 ml / kg) was orally administered on the first day of the experiment (day 0), and after 30 minutes, 27 gages were used. 15 μl (15 nmol) of 1 mM Aβ (25-35) was slowly injected into the ventricle using a microsyringe equipped with a needle. The same amount of physiological saline for injection was administered to the Siamese control group in the same manner. Thereafter, three weeks or more of the adaptation tests were performed in which the animals were placed in bright boxes and orally administered with rice bran oil once daily for one week. Corn oil was orally administered to the control group that did not administer Aβ (25-35) and the control group that did not administer Aβ (25-35) but did not receive rice bran extract. On day 7, the rice bran oil was orally administered, and after 30 minutes, the animals were placed in bright boxes, and after 10 seconds, the doors were opened, and when the animals entered the dark boxes, the doors were closed and electric stimulation (0.2 mA, 2 seconds) was applied. . The animal thus remembers the experience of electric shock in a dark box. After 24 hours, the animal is placed in a bright box, and the same experiment is carried out. The animal that remembers the shock of the previous day does not move to the dark box. Therefore, the animal staying time in the bright box is measured as an indicator of memory formation, and the staying time of 5 minutes or more is 5 minutes.

As a result, as can be seen in Figure 8, one week after the Aβ (25-35) (15 nmol) administered in the ventricle and passive avoidance test was compared with the control group Aβ ( 25-35) only moved to the dark room at 51 seconds. This means that no memory is formed. In contrast, those who received 0.5 and 1 ml / kg of rice bran extract daily for 1 week were 186 seconds for 0.5 ml / kg of rice bran extract and 229 seconds for 1 ml / kg dose group. All groups were significantly increased to inhibit memory damage caused by Aβ (25-35).

From the above results, it was confirmed that the rice bran oil extract can exhibit an excellent memory damage inhibitory effect in a dose-dependent manner.

[ Experimental Example  3] Measurement of neuronal cell death inhibitory effect- MTT Color Assay

1) Culture of Cerebral Cortical Neurons

Rats at 15 days of gestation were removed from the fetus under ether anesthesia and only cerebral cortex was isolated under a microscope. This was added to Joklik-modified Eagle's medium containing trypsin (0.25 mg / ml) and mechanically dispersed by 5 ml pipette. This was incubated at 37 ℃ for 10 minutes to perform enzymatic separation. The cell suspension was centrifuged at 1,500 rpm for 5 minutes in a precipitate layer containing the cells sodium bicarbonate (44 mM), penicillin (40 U / mL), gentamycin (50 g / mL), KCl (5 mM) and 10 A culture vessel coated with poly-L-lysine after addition of DMEM containing% fetal bovine serum (FBS) to adjust the cell concentration to 2 × 10 ⁶ cells / ml, passing through a nylon mesh (35 μm) Planted in. Incubation was carried out in a CO ₂ incubator at 37 ° C. and 5% CO ₂ /95% air.

2) MTT Assay (Measure viable cell number)

In order to elucidate the mechanism explaining the inhibitory effect of rice bran oil on cerebral infarction, experiments were carried out using the cultured neuronal neurons of the above-mentioned strain. All experiments were measured for cell viability by MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl-tetrazolium bromide) assay. This method is a very sensitive and stable method for quantitatively measuring the survival and proliferation of cells, based on the mitochondria's activity in converting the yellow water-soluble substrate MTT into dark blue formazan in living cells. Therefore, the amount of formazan produced is proportional to the number of living cells, and the higher the absorbance, the more the number of living cells.

After removing the medium of the cells cultured for a certain time after drug treatment, MTT (0.5 mg / ㎖) solution was added and incubated at 37 ℃ for 4 hours. The MTT solution was then removed and 200 μl of acid-isopropanol (0.04 N HCl in isopropanol) was added to all wells to dissolve the dark blue formazan crystals formed, followed by a wavelength of 570 nm with a microelisa reader. The absorbance was measured by (reference wavelength 630 nm). At this time, when the cells were not treated with the drug (control) as a control (100%), the absorbance of glutamate, hypoxia, Aβ (25-35), and a group of rice bran extracts treated with them were expressed in%.

(1) Aβ (25-35) -induced Neuronal cell death  About Rice Bran Oil  Determination of Inhibitory Effect of Extracts

Aβ (25-35) was administered to the ventricles of mice to cause memory damage. In contrast, chronic administration of rice bran oil orally inhibited memory damage. In order to investigate the biochemical mechanism, the cerebral cortical neurons were cultured and the cell death by 10 μM Aβ (25-35) was induced and the inhibitory effect of rice bran oil was investigated.

As a result, as shown in FIG. 9A, when treated with 10 μM Aβ (25-35), the Aβ (25-35) treated group had 56.1% less viable cells compared to the control group (100%). Treatments treated with the cultured neuronal cells were confirmed to recover the survival rate by 59.7, 67.5, 78.0% by 0.1, 1, 5 μl / ㎖ respectively.

(2) Glutamate -Judo Neuronal cell death  About Rice Bran Oil  Determination of Inhibitory Effect of Extracts

In degenerative brain diseases such as cerebral infarction, it is known that glutamate, an excitatory amino acid, is released in large amounts, causing neuronal death. Therefore, glutamate (500 μM) was added to the cultured cerebral cortical neurons of 1) and cultured for 14 hours to investigate the inhibitory effect of rice bran oil on neuronal cell death.

As a result, as shown in B of FIG. 9, the cell viability of the glutamate treatment group was reduced to 73.6% compared to the control group (100%), whereas the rice bran extract was extracted in 79.1% and 1 μl / ml by 0.1 μl / ml. As a result, it was confirmed that the survival rate of 84.2% was obtained by 88.7% and 5 μl / ml. In particular, 1, 5 μl / ml dose of rice bran extract significantly inhibited cultured neuronal cell death by glutamate.

(3) hypoxia ( hypoxia )-Judo Neuronal cell death  About Rice Bran Oil  Determination of Inhibitory Effect of Extracts

In order to satisfy the condition of ischemia, the neuronal cells of the cultured cerebral cortex of 1) were cultured in a chamber for hypoxia condition to induce cell death and to investigate the inhibitory effect of rice bran oil extract. In other words, the cultured cerebral cortical neurons were cultured on the 5th to 7th day of culture, and the medium was changed to glucose-deficient HEPES buffer and then plated in a hypoxic chamber maintained at 2% O ₂ , 5% CO ₂ , and 93% N ₂ . Was added and incubated for 24 hours in a chamber. The medium was then exchanged with serum-free DMEM and incubated for another 6 hours in a 5% CO ₂ incubator and cell viability was measured via MTT assay. As a control, cells substituted with serum-free DMEM and glucose-deficient HEPES buffer were exposed to the 5% CO ₂ incubator for the same time.

As a result, as shown in FIG. 9C, when the cell survival of cells cultured in normal medium was 100%, cells cultured in glucose-deficient HEPES medium showed 83.7% survival rate. Survival was 63.5% at 24 hours culture and confirmed about 20% cell death. In the treatment group treated with rice bran oil extract 0.1, 0.5, 1 μl / ml showed 66.9, 68.5, 75.1% of brain cell viability, and 1 μl / ml treatment group showed significant cell death inhibition.

From the above results, it can be seen that the rice bran oil extract according to the present invention can be effectively used as a preventive or therapeutic agent for cerebrovascular diseases such as cerebral infarction, cerebral ischemia, stroke, and vascular dementia.

Claims (7)

A composition for preventing or treating memory damage, comprising a rice bran oil extract obtained by shaking extraction with an organic solvent from rice bran oil as an active ingredient. The method of claim 1,
The rice bran oil extract is a mixture for thermocompression rice bran oil and hexane in a weight ratio of 1: 3 to 5, and then extracting shaking at a temperature of 30 to 50 ℃ memory composition for preventing or treating damage. The method of claim 2,
The thermally compressed rice bran oil is a composition for preventing or treating memory damage, characterized in that the product is prepared by pressing the rice bran at a pressure of 50 to 70 Mpa at a temperature of 70 to 80 ℃. The method according to any one of claims 1 to 3,
The memory damage is a memory damage prevention or treatment composition, characterized in that the memory damage by Aβ factor. 1) thermocompressing rice bran to obtain rice bran oil;
2) extracting the obtained rice bran oil with an organic solvent; And
3) drying the extracted rice bran oil;
Method for producing a composition for preventing or treating memory damage, including the production. 6. The method of claim 5,
Thermocompression of step 1) is a method of producing a composition for preventing or treating memory damage, characterized in that the compression at 50 to 70 Mpa pressure at a temperature of 70 to 80 ℃. 6. The method of claim 5,
The extraction of step 2) is a method of producing a composition for preventing or treating memory damage, characterized in that after mixing the rice bran oil and the organic solvent in a 1: 3 to 5 weight ratio, shaking extraction at a temperature of 30 to 50 ℃.

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