KR102296272B1 - A composition for improving memory and cognitive function, preventing and improving ischemia reperfusion injury including acorus gramineus soland extract - Google Patents

Abstract

The present invention relates to a composition for improving memory and cognitive function, preventing and improving ischemic disease or ischemic reperfusion-related disease, comprising a seokchangpo extract. The present invention provides a composition comprising a seokchangpo extract having an effect of improving memory and cognitive function based on a natural product. In addition, the present invention provides a composition comprising a seokchangpo extract having an effect of preventing and improving ischemic reperfusion injury.

Description

A composition for improving memory and cognitive function, preventing and improving ischemia-reperfusion injury, comprising an extract of oleander

The present invention relates to a composition for improving memory and cognitive function, and preventing and improving ischemia-reperfusion injury, comprising a seokchangpo extract. More specifically, it is to provide a composition capable of preventing and ameliorating a disease caused by ischemic reperfusion injury by preventing or alleviating memory and cognitive function improvement activity and ischemic reperfusion injury.

Cerebral infarction and myocardial infarction, which are typical ischemic diseases, are in the state of arteriosclerosis, which narrows blood vessels due to high blood pressure, hyperlipidemia, diabetes, and smoking. It is caused by tissue necrosis.

In particular, stroke can be largely divided into cerebral ischemia, which occurs when blood supply is blocked due to cerebral blood vessel stenosis or blockage by a thrombus, and cerebral hemorrhage due to intracranial hemorrhage. Among them, cerebral ischemia is caused by a lack of oxygen and nutrients due to blockage of blood supply, leading to a decrease in ATP in nerve cells and edema, which directly induces cell damage and necrosis. Such edema interferes with the smooth functioning of the brain and induces tissue necrosis, leading to extensive brain tissue damage, resulting in secondary behavioral or cognitive impairments depending on the damaged area. Therefore, most stroke patients have behavioral disorders due to brain tissue damage even after the thrombus is removed, which is also an important part of stroke treatment. In addition, recently, it is a disease that occurs frequently enough to be the number one cause of death. Currently, the treatment for cerebral ischemia is t-PA (tissue plasminogen activator) approved by the FDA. , it cannot be used when there is a possibility of hemorrhagic brain ischemia at the same time.

On the other hand, ischemia refers to a decrease in local blood flow in a state where oxygen supply is insufficient than the oxygen demand required for physiological functions. Tissue damage is further accelerated due to reoxidation of tissues that tend to be highly susceptible to oxidative damage during the This is called ischemia/reperfusion injury (IRI).

Ischemia-reperfusion injury is well known to cause pathological conditions. It may indicate a predicted morbidity or an unexpected morbidity. The aforementioned stroke is one of the most common forms of unpredictable ischemia-reperfusion injury, which is the third leading cause of death in industrialized countries and a leading cause of long-term disability. Stroke is a type of cardiovascular disease that affects the arteries within and to the brain. A stroke occurs when those arteries are blocked by a clot, resulting in ischemia of the cerebral tissue due to the blocked artery. Direct damage to the brain is caused by obstruction of blood flow, mainly due to lack of oxygenation in living tissue, eventually resulting in infarction if left unaltered. However, if such damage is altered (physiologically or therapeutically), then reperfusion of the ischemic tissue can paradoxically cause additional indirect damage. When ischemia persists for a long time, "direct" damage from oxygen deprivation alone is the main mechanism. For short duration of ischemia, indirect or reperfusion mediated injury is becoming increasingly important to the end result.

However, as a treatment method for ischemia developed so far, it is best to reperfusion the blood vessel by piercing the blocked blood vessel in the kidney where ischemia has occurred in a short time with a thrombolytic agent, etc. It is not expected, and additional damage caused by reperfusion is unavoidable, and it also delays the recovery of the transplanted kidney during kidney transplantation, increases acute rejection, and even reduces the survival rate of transplanted kidney in the long term.

That is, in the case of ischemia-reperfusion injury that can occur in various surgical operations, there is still no therapeutic agent that can be applied clinically. development is imperative. Accordingly, studies to elucidate substances and mechanisms caused by ischemia in the brain, cardiovascular, and liver are continuing, but the results are insufficient.

Therefore, there is a demand for the development of a material that can more effectively prevent and improve the ischemic reperfusion injury.

KR 1994-7003658 A KR 10-2008-0098001 A

It is an object of the present invention to provide a composition comprising a seokchangpo extract having an effect of improving memory and cognitive function based on a natural product.

It is an object of the present invention to provide a composition comprising a seokchangpo extract having an effect of preventing and improving ischemic reperfusion injury.

The composition comprising the extract according to the present invention is a natural extract, and there is no problem of side effects such as cytotoxicity.

It is an object of the present invention to provide a composition for improving memory and cognitive function, and preventing and improving ischemic reperfusion injury, including the extract of seokchangpo, in the form of food, health functional food, and the like.

In order to achieve the above object, the composition for improving memory and cognitive function according to an embodiment of the present invention may include a seokchangpo extract.

In the composition for improving memory and cognitive function, the extract may be one selected from the group consisting of water, alcohols having 1 to 10 carbon atoms, and mixtures thereof with a solvent.

The composition for preventing and improving ischemic reperfusion injury according to another embodiment of the present invention may include a seokchangpo extract.

In the composition for preventing and improving ischemic reperfusion injury, the extract may be one selected from the group consisting of water, alcohols having 1 to 10 carbon atoms, and mixtures thereof with a solvent.

A food for improving memory and cognitive function according to another embodiment of the present invention includes the composition, Schisandra extract, strawberry extract, acai berry extract, aronia extract, blueberry extract, cranberry extract, grapefruit extract, orange extract, Selected from the group consisting of lemon extract, chocomint extract, basil extract, peppermint extract, sage extract, lavender extract, parsley extract, gompi extract, beetroot extract, coltsfoot extract, soothing extract, green tea extract, noni extract, and mixtures thereof It may further include any one.

A food for preventing and improving ischemic reperfusion injury according to another embodiment of the present invention includes the above composition, and includes Schisandra extract, strawberry extract, acai berry extract, aronia extract, blueberry extract, cranberry extract, grapefruit extract, and orange Extracts, lemon extract, chocomint extract, basil extract, peppermint extract, sage extract, lavender extract, parsley extract, gompi extract, beetroot extract, coltsfoot extract, soothing extract, tea extract, noni extract and mixtures thereof It may be to further include any one selected from.

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

The composition for improving memory and cognitive function according to an embodiment of the present invention may include a seokchangpo extract.

The above-mentioned Seokchangpo (石菖蒲) is a perennial herb with green leaves all year round. The rhizome is thick and hard, has many nodes, and gives off fine roots and lives in the same place as a rock crevice. The leaves grow from the rhizome in a state where the bases overlap each other. Narrow lines are tough and shiny. The tip of the leaf is sharp like a knife, the edge is flat, and it always smells good. Flowers are clustered in a round stick shape in the middle of a flower stalk that has a leaf-like shape. The length of the flower stick is about 5cm and the color is yellowish blue.

As used herein, the term "extract" has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the seokchangpo extract includes not only those obtained by using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the extract of Seokchangpo of the present invention.

The seokchangpo extract may exhibit the effect of improving memory and cognitive function and preventing or improving ischemic disease.

Preferably, the ischemic disease may be a stroke or cerebral infarction.

Preferably, the composition for improving memory and cognitive function may include extracts from stock extracts, gompi extracts, beech extracts, and chazugi extracts. In the case of the above range, the activity of improving memory and cognitive function may be increased due to a synergistic effect due to the interaction.

More preferably, the composition for improving memory and cognitive function may further include 1 to 5 parts by weight of gompi extract, 10 to 20 parts by weight of perennial herbs, and 5 parts by weight of chazugi extract 1 based on 100 parts by weight of the seokchangpo extract. . According to the above range, it is possible to provide not only excellent memory and cognitive function improvement activity, but also a formulation capable of improving taste and flavor when provided as a food composition. On the other hand, in the case of containing the base paper extract, memory and cognitive function improvement activity is excellent, but when it is provided as a food composition, it is not possible to suppress the palatable low flavor and taste of the base paper even by mixing other natural substances described later, so it can be provided as a palatable food composition. There is a problem that you can't.

In the composition for improving memory and cognitive function, the extract may be one selected from the group consisting of water, alcohols having 1 to 10 carbon atoms, and mixtures thereof with a solvent.

Preferably, the solvent may be water. When water is used as a solvent to prepare a seokchangpo extract, it may be advantageous to obtain an active substance.

The composition for preventing and improving ischemic reperfusion injury according to another embodiment of the present invention may include a seokchangpo extract.

Rapid reperfusion is essential to suppress tissue damage caused by ischemia, such as blood vessel blockage and narrowing. However, in the process of blood reperfusion, additional damage is accompanied by aggravating tissue damage, such as decreased vascular endothelial cell function. This is called ischemia reperfusion injury (IRI). In particular, when ischemia occurs in tissues or organs, damage is unavoidable, and the extent and degree of damage are determined according to the time of reperfusion. If the ischemia time is prolonged, irreversible damage occurs, and even if reperfusion occurs, severe damage can be caused by reperfusion.

In the present invention, "ischemic damage" refers to damage caused by reduced oxygen delivery while blood circulation is blocked in organs that require blood flow, such as the heart, brain, and kidney, and can lead to tissue functional damage and cell death. includes damage. Possible causes of ischemic injury include vascular disease, coronary thrombosis, cerebrovascular thrombosis, aneurysm rupture, systemic hemorrhage, crush injury, sepsis, severe skin burns, and vascular occlusion surgical procedures (e.g., spinal cord during thoracoabdominal aneurysm surgery). ischemia), cardiopulmonary bypass method, organ transplantation, cardiopulmonary collapse (acute cardiac death), and asphyxiation.

In the present invention, "ischemic injury" includes ischemic reperfusion injury that may occur due to organ transplantation, etc. in addition to ischemic injury that may normally occur.

In the present invention, "ischemic reperfusion injury" refers to myocardial ischemia, cerebrovascular ischemia, renal ischemia, liver ischemia, ischemia-reperfusion cardiomyopathy, skin ischemia, intestinal ischemia, intestinal ischemia, gastric ischemia, pulmonary ischemia, pancreatic ischemia, skeletal muscle ischemia, abdominal muscle ischemia. , extremity ischemia, ischemia-reperfusion colitis, mesenteric ischemia, and asymptomatic ischemia may be caused by at least one ischemia selected from the group consisting of, but not limited to.

The ischemic reperfusion injury is the restoration of blood flow in a tissue or organ that has suffered a decrease in blood supply or a natural event, such as restoration of blood flow after trauma, eg, acute myocardial infarction, or restoration of blood flow in a tissue or organ that has undergone reperfusion surgery, such as a decrease in blood supply. It can be caused by one or more surgical procedures, other therapeutic interventions, or organ transplant procedures. The surgical procedure includes, for example, coronary artery bypass grafting, coronary angioplasty, and organ transplantation.

For example, acute myocardial infarction refers to a phenomenon in which the heart muscle is killed or necrotic because one of the blood vessels of the heart is occluded and oxygen and nutrients are not supplied. The mortality rate can be reduced to less than 10%. However, in about 10% of patients, the clogged blood vessels are reperfused and the myocardial cells are damaged due to the rapid oxygen supply. This is called reperfusion injury. % will develop symptoms such as heart failure.

The cerebrovascular disease includes, but is not limited to, stroke, cerebral infarction, cerebral thrombosis, cerebral embolism, and the like. The cardiovascular disease includes, but is not limited to, myocardial infarction, angina pectoris, and the like.

The seokchangpo extract may exhibit the effect of preventing or alleviating ischemia-reperfusion damage by inhibiting the mitochondrial-dependent apoptotic pathway. Accordingly, it is possible to prevent and improve diseases caused by ischemia-reperfusion injury.

Preferably, the composition for the prevention and improvement of ischemic reperfusion injury may include a stock extract, a gompi extract, an extract of fenugreek tree, and an extract of chazugi. In the case of the above range, the effect of preventing and improving ischemia reperfusion damage by preventing mitochondrial and apoptosis due to a synergistic effect by interaction can be increased.

More preferably, the composition for preventing and improving ischemic reperfusion injury further comprises 1 to 5 parts by weight of Gompi extract, 10 to 20 parts by weight of beetroot extract, and 5 parts by weight of Chazugi extract 1 based on 100 parts by weight of the seokchangpo extract. can According to the above range, the effect of preventing and improving ischemic reperfusion damage as an effect of inhibiting antioxidant activity and mitochondrial damage or apoptosis may be further improved. Accordingly, it is possible to exhibit high activity with a smaller content.

In the composition for preventing and improving ischemic reperfusion injury, the extract may be one selected from the group consisting of water, alcohols having 1 to 10 carbon atoms, and mixtures thereof with a solvent.

Preferably, the solvent may be water. When water is extracted with a solvent, it may be advantageous to obtain an active substance.

The food for improving memory and cognitive function according to another embodiment of the present invention includes the composition, Schisandra extract, strawberry extract, acai berry extract, aronia extract, blueberry extract, cranberry extract, grapefruit extract, orange extract, Any from the group consisting of lemon extract, chocomint extract, basil extract, peppermint extract, sage extract, lavender extract, parsley extract, gompi extract, beetroot extract, coltsfoot extract, soothing extract, chazu extract, noni extract, and mixtures thereof It may include one more.

The food is to include general food, processed food, health functional food, functional food, illustratively as a formulation for the food can be applied to beverages, tea, tablets, processed meat food, snacks, instant food, etc. .

A food for preventing and improving ischemic reperfusion injury according to another embodiment of the present invention includes the above composition, and includes Schisandra extract, strawberry extract, acai berry extract, aronia extract, blueberry extract, cranberry extract, grapefruit extract, and orange Extracts, lemon extract, chocomint extract, basil extract, peppermint extract, sage extract, lavender extract, parsley extract, gompi extract, beetroot extract, coltsfoot extract, soothing extract, tea extract, noni extract and mixtures thereof It may be to further include any one in.

Preferably, the food or health functional food is Omija 0.1 to 1 parts by weight based on 100 parts by weight of Seokchangpo extract; 0.1 to 1 part by weight of cranberry extract; It may further include 0.1 to 2 parts by weight of peppermint extract, 1-3 parts by weight of parsley extract, and 0.1 to 1 parts by weight of soothing extract. According to the above range, it is possible to provide a food composition with a palatable texture with a fairly deep and fresh flavor while neutralizing the low palatability and taste of the extract of seokchangpo. In particular, when according to the above range, it is included in less than 10% by weight compared to the seokchangpo extract, and improves the taste and flavor of the seokchangpo extract as described above without compromising the functionality of the seokchangpo extract to provide a food composition with a fairly high palatability. can In addition, the above-mentioned Gompi extract; Even in the case of adding a persimmon leaf extract and chazugi extract, the taste and flavor are excellent, so the palatability is high, and it has the advantage that the activity of the active ingredient desired in the present invention can be maintained as much as possible.

The health functional food is not particularly limited as long as it can be ingested to improve cognitive function or memory, prevent or improve ischemic disease, or prevent or improve ischemic reperfusion injury.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The active ingredient may be used appropriately depending on the purpose of its use (prevention or improvement). In general, in the production of food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for health purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the type of the health functional food. The health functional food composition of the present invention may be prepared as a food, particularly a functional food. The functional food of the present invention is

It includes ingredients commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.) or sugar alcohols (eg, , xylitol, sorbitol, erythritol, etc.). As the flavoring agent, natural flavoring agents (eg, taumatine, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used.

In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid It may further contain a carbonation agent and the like used in beverages.

The present invention provides a composition comprising a seokchangpo extract having the effect of improving memory and cognitive function, and preventing and improving ischemic disease based on a natural product.

The present invention provides a composition comprising a seokchangpo extract having an effect of preventing and improving ischemic reperfusion injury.

The composition comprising the extract according to the present invention is a natural extract and has no side effects such as cytotoxicity, so that it can be used for a long period of time, improving memory and cognitive function, and preventing and improving ischemic reperfusion injury.

The present invention can provide a composition for improving memory and cognitive function, preventing and improving ischemic reperfusion injury, including the extract of seokchangpo, in the form of food, health functional food, and the like.

1 relates to cell viability in an OGD model according to an embodiment of the present invention.
2 is for the evaluation of LDH expression in the OGD model according to an embodiment of the present invention.
3 is a change in reactive oxygen species in the OGD model according to an embodiment of the present invention.
4 is a change in reactive oxygen species in the OGD model according to an embodiment of the present invention.
5 is a change in reduced glutathione (GSH) in the OGD model according to an embodiment of the present invention.
6 illustrates changes in intracellular active oxidative species due to oxidative stress in the OGD model according to an embodiment of the present invention.
7 is related to the evaluation of anti-inflammatory activity in the OGD model according to an embodiment of the present invention.
8 is related to the evaluation of inhibitory activity of ischemic stroke in the OGD model according to an embodiment of the present invention.
9 relates to a cytoprotective activity according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Preparation Example 1: Preparation of extract of seokchangpo extract]

After washing, drying, crushing, hot water extraction, and concentrated filtration, Seokgampo extract (WAGS) was prepared. The following experiment was performed while diluting the Seokchangpo extract to a certain concentration.

[Experimental Example 1: Prevention and improvement activity of ischemic relapse duct damage through OGD model]

1. Construction of OGD Model and Cell Culture

In order to confirm the effect of alleviating ischemia-reperfusion injury of the extract according to the preparation example, an in vitro model of ischemia was established through an oxygen-glucose deprivation (OGD) test method in cells. On the other hand, human neuroblastoma SH-SY5Y cell (ATCC no. CRL-2266) is dulbecco's modified eagle's medium (Sigma-Aldrich) containing 10% fetal bovine serum (Gibco-BRL, USA) and antibiotics (Sigma-Aldrich, USA). , USA) cultured under medium. The incubator was maintained at a temperature of 37° C., and _{a gas mixture of 95 v% air and 5 v% CO 2} was continuously supplied to prepare suitable conditions for cell culture.

2. Analysis of cell viability of seokchangpo extract

To analyze the cell viability, treated human neurons (SH-SY5Y) ^{were inoculated into a 6-well plate at 3 x 10 5} cells/ml, cultured for 24 hours, and treated with various concentrations of WAGS. and then incubated for 24 hours.

As a result, as a result of treatment with WAGS in the range of 10 to 500 μg/ml, it was confirmed that WAGS had no significant effect on cell viability.

3. Analysis of oxygen-glucoside privation (OGD) and cell viability and LDH

OGD-induced apoptosis is a method of inducing cell death by temporarily blocking the supply of nutrients and oxygen, causing oxidative stress in cells, and re-supplying nutrients and oxygen. it can be said Reactive oxygen species can directly induce cell damage or indirectly cause cell damage through signaling systems. These reactive oxygen species play an etiologically important role in several central nervous system diseases including stroke and various degenerative brain diseases. In order to artificially stop the supply of glucose and serum to human neuroblastoma SH-SY5Y cells and create an oxygen-deficient environment to induce apoptosis, the OGD model sets an environment similar to the ischemic state following the interruption of blood supply. , while inducing brain neuronal cell death according to ischemic conditions, the cell viability according to WAGS administration was evaluated. Referring to FIG. 1 for the results, it can be seen that the cell viability is improved in a concentration-dependent manner when WAGS is treated even in the OGD model.

Meanwhile, serum lactate dehydrogenase (LDH) activity in the OGD model was measured and confirmed by ELISA. LDH is a representative index component for ischemic reperfusion injury. Referring to FIG. 2 below, it can be seen that the expression level of LDH is decreased in a concentration-dependent manner when WAGS is treated.

4. Measurement of Enzyme Activity

In the OGD model, the TBARS method was used to measure malondialdehyde (MDA) in order to investigate the change of intracellular reactive oxygen species due to external oxidative stress according to WAGS. To evaluate the activity of superoxide dismutase (SOD), SOD Assay Kit was used, and the activity of glutathione peroxidase (GPx) was measured using Glutathione peroxidas cellular Activity Assay Kit.

The results are shown in FIGS. 3 to 5 below. 3 to 5, it can be confirmed that WAGS can significantly reduce the concentration of oxidative stress induced by the OGD model.

In addition, in the OGD model, dichlorodihydro-fluorescein diacetate (DCF-DA), which is deacetylated and fluorescing when combined with reactive oxygen species, was used to examine changes in intracellular reactive oxygen species due to oxidative stress. was measured. It is shown in Figure 6 below. Referring to FIG. 6, it can be seen that the oxidative stress induced by the OGD model in WAGS decreases in a concentration-dependent manner.

5. Anti-inflammatory activity evaluation

Cytokines are substances that control the immune response, infection, trauma, inflammation, etc. and transmit signals between cells. Pro-inflammatory cytokines include IL-1 (IL-1α, β), IL-6, IL-8, TNF-α, G-CSF, GM-CSF, interferon (IFN)-γ, CXCL1, CXCL2, etc. It plays an important role in inducing and enhancing and may cause damage.

In particular, the inflammatory response is known to be one of the factors of ischemia-reperfusion injury. Therefore, in the OGD model, the expression of NO and inflammatory cytokines according to the seokchangpo extract (WAGS) was evaluated and shown in FIG. 7 below.

Referring to FIG. 7 below, it can be seen that the concentration-dependent anti-inflammatory activity is increased in the OGD model in the case of the seokgyepo extract (WAGS). Accordingly, it can be confirmed that the ischemia-reperfusion injury caused by inflammation can be prevented or improved by the sekkipo extract (WAGS).

5. Evaluation of inhibitory activity of ischemic stroke

Ischemic stroke is an important factor in the pathogenesis of vascular dementia and Alzheimer's dementia.

Aβ, which is excessively deposited in the form of plaques in the brain of these dementia patients, and is pointed as the cause of brain tissue degeneration, is generated by sequentially acting on APP by β-secretase and γ-secretase. In particular, the plaque is mainly composed of Aβ consisting of 40 to 42 amino acids produced by cleaving the APP protein.

There are two main types of Aβ, of which Aβ ₄₀ increases with aging in normal brains, but Aβ _42, which has high neurotoxicity, increases specifically in the brains of dementia patients. The production of _{Aβ 40} and Aβ ₄₂ is directly determined by γ-secretase cleavage. On the other hand, the expression of β-secretase is increased in the brain tissue of dementia patients. In particular, the increase in the expression of β-secretase due to oxidative stress is very distinct, and the expression of β-secretase by this oxidative stress is dependent on the activity of γ-secretase.

Therefore, in the OGD model, the production of Aβ ₄₀ and Aβ ₄₂ peptides, and the expression levels of β-secretase and γ-secretase were evaluated while processing the extract of seokchangpo (WAGS), and it is shown in FIG. 8 below.

Referring to FIG. 8 below, in the case of WAGS in the OGD model, it can be seen that the production amount _{of Aβ 40} and Aβ _{42 is decreased in a concentration-dependent manner.} Also, it can be seen that the expression of β-secretase and γ-secretase is decreased. Accordingly, it was confirmed that the occurrence of vascular dementia or Alzheimer's dementia could be prevented and improved through prevention or improvement of ischemic stroke in the case of the seokchangpo extract (WAGS).

6. Apoptotic cell death

Cell apoptosis was performed by flow cytometry (BD, Mountain View, USA). The results are shown in FIG. 9 below. Referring to FIG. 9 below, it can be confirmed that neuronal cell death is prevented when WAGS is treated in the OGD model.

In the end, the seokchangpo extract (WAGS) has resistance activity to external oxidative stress in human nerve cells (SH-SY5Y) and inhibits mitochondrial-dependent apoptosis pathway by inhibiting mitochondrial damage and apoptosis. It was confirmed that the ischemic reperfusion injury could be alleviated.

[Preparation Example 2: Preparation of mixed extract]

Through hot water extraction according to the same method as above and WAGS, Gompi extract (SGP), Windbreak extract (SDS), Chazugi extract (SPF), Raw paper extract (SPT), Schisandra extract (E1), Cranberry extract (E2) , peppermint extract (E3), parsley extract (E4) and soothing extract (E5) were prepared, and a mixed composition was prepared according to the composition of each extract as shown in [Table 1] below.

M1 M2 M3 M4 M5 M6 T1 T2 T3 T4 T5 T6 WAGS 100 100 100 100 100 100 100 100 100 100 100 100 SGP 0.1 One 3 5 10 - - - - - - 3 SDS One 10 15 20 40 - - - - - - 15 SPF 0.1 One 3 5 10 - - - - - - 3 SPT - - - - - 20 - - - - 20 - E1 - - - - - - 0.01 0.1 One 3 One One E2 - - - - - - 0.1 One 3 5 3 3 E3 - - - - - - 0.01 0.1 2 5 2 2 E4 - - - - - - One One 3 5 3 3 E5 - - - - - - 0.1 0.1 2 5 2 2

(Unit: parts by weight)

[Experimental Example 2: Memory and cognitive function improvement effect experiment]

1. DPPH radical analysis

For the analysis of reactive oxygen species such as DPPH and superoxide anion related to oxidative stress in vivo, the radical scavenging ability of WAGS and M1 to M6 was measured using 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical. did. After reacting by adding a DPPH solution to WAGS and M1 to M6, absorbance was measured. The DPPH radical content is shown in [Table 2] by converting the absorbance ratio of the sample addition group and the control group to a % value.

WAGS M1 M2 M3 M4 M5 M6 DPPH radical (%) 59 61 49 41 48 58 40

2. Reducing Power Analysis

Superoxide anion radical scavenging activity was performed according to the Fridovich method. To 1 ml of the sample, 200 mM phosphate buffer of pH 6.6 and 1 ml of potassium ferricyanide were sequentially added and reacted. Here, 10% TCA (trichloroacetic acid) solution was mixed with distilled water and ferric chloride in the supernatant, and absorbance was measured. WAGS and M1 to M6 are shown in [Table 3] below by converting the absorbance ratio into a % value based on the value of the control group.

WAGS M1 M2 M3 M4 M5 M6 Superoxide anion (%) 258 269 338 377 325 237 324

3. Analysis of DNA damage by hydroxyl radical

Genomic DNA was extracted from PC12 cells according to a slightly modified standard procedure. WAGS and M1 to M6, 200 μM FeSO4, 1 mM H2O2, and 50 μg/ml genomic DNA were added to the DNA solution. After completion of the reaction, electrophoresis was performed. After staining, it was observed using a LAS3000 image analyzer with UV, and the results were expressed in % based on the untreated group and shown in [Table 4] below, and the lower the % value, the better.

WAGS M1 M2 M3 M4 M5 M6 DNA oxidation(%) 69 65 45 49 52 63 53

4. Measurement of Angiotensin I Converting Enzyme (ACE) Activity

ACE activity was pre-reacted by adding sodium borate buffer (pH 8.3) and ACE coenzyme solution to WAGS and M1 to M6, respectively, and then reacted by adding a substrate prepared by adding HHL (hippuryl-histidyl-leucine). Meanwhile, the ACE coenzyme solution was prepared by extracting rabbit lung acetone powder in sodium borate buffer (pH 8.3) and then centrifuging. The supernatant collected after the reaction was mixed with dried and distilled water, and absorbance was measured. Distilled water was used as a negative control group, and the absorbance ratio before and after addition of the sample was converted into a % value based on the untreated group as a standard (100%), and is shown in [Table 5] below.

WAGS M1 M2 M3 M4 M5 M6 ACE inhibition (%) 230 227 257 264 254 224 266

5. AChE (Acetylcholine esterase) activity measurement

50 μl of AchE (25 mU/ml) isolated from PC12 cells was used as a reaction enzyme, and acetylcholine iodide dissolved in 0.1 M sodium phosphate buffer (pH 7.8) was used as a substrate, and the chromogenic reagent was 5,5'-dithiobis-2. -nitrobenzoic acid and sodium hydrogen carbonate were dissolved in 10 ml of 0.1 M sodium phosphate buffer (pH 7.0). After the reaction, the absorbance value was measured at 412 nm with a spectrophotometer. The activity of AChE is shown in [Table 6] below by converting the absorbance ratio before and after addition of WAGS and M1 to M6 into % values based on the untreated group (100%).

WAGS M1 M2 M3 M4 M5 M6 AChE activity (%) 56 57 48 49 48 61 50

6. Test for NF-κB transcriptional activity in C12 cells

While culturing PC12 cells, the transcriptional activity of NF-κB was evaluated using the NF-κB promoter-luciferase construct, and the effect on NF-κB transcriptional activity in C12 cells for WAGS and M1 to M6 was compared. The result of WAGS was set to 10, and the results of M1 to M6 were evaluated as indices, and are shown in [Table 7]. The higher the index, the better the transcriptional activity of NF-κB.

WAGS M1 M2 M3 M4 M5 M6 Relative luciferase activity 10 10 16 16 14 9 14

In the end, according to the present invention, removal of reactive oxygen species, antioxidant against DNA damage, ACE (Angiotensin Converting Enzyme) enzyme inhibitory activity, AChE (acetylcholine esterase) inhibitory activity, NO (nitric oxide) production inhibitory activity, NF-κB transcription It can be seen that the activity increasing effect appears. In addition, it can be seen that a certain synergistic effect appears in the case of the mixed composition of M2 to M4 and M6.

Therefore, it can be applied to a variety of products that can produce effects for improving memory and brain cognitive function based on the mixed composition of the seokchangpo extract or the seokchangpo extract.

[Experimental Example 3: Evaluation of palatability]

A tea beverage was prepared according to the composition of WAGS and T1 to T5, and after tasting it, 10 testers were asked to evaluate the flavor and taste to evaluate the preference of 1 to 10. The results are summarized and averaged and shown in [Table 8] below. In the following indices, the higher the number, the better the preference (rounding is applied to the average value for each index).

WAGS T1 T2 T3 T4 T5 T6 palatability One One 6 6 3 One 7

(Unit: Index)

As a result of the palatability evaluation, most of the tasters rated the palatability for the aroma and taste of Seokchangpo as low. However, in the case of the mixed composition of T2 and T3, it was confirmed that the overall taste and flavor were excellent while the sweet and refreshing scent and the unique flavor of seokchangpo were subtly felt. In particular, in the case of the mixed composition, since the mixture of E1 to E5 is contained in less than 10% by weight based on the extract of Seokchangpo, the palatability can be excellent while maintaining the activity according to the active ingredient of the Seokchangpo as much as possible. In addition, it can be seen that, in the case of the above composition, an effect of improving palatability can be obtained even in the composition for M3.

On the other hand, in the case of T5, even when a mixture of E1 to E5 is used, the bitter taste of the base paper itself is not neutralized and the palatability is quite low, so it may be difficult to provide a palatable food composition by mixing a small amount of plant extract.

Although the preferred embodiment of the present invention has been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements by those skilled in the art using the basic concept of the present invention as defined in the following claims are also provided. is within the scope of the

Claims (3)

Contains seokchangpo extract and natural flavoring agent,
The natural flavoring agent includes omija extract, cranberry extract, peppermint extract, parsley extract and soothing extract,
The natural flavoring agent is 0.1 to 1 parts by weight of Schisandra extract, 1 to 3 parts by weight of cranberry extract, 0.1 to 2 parts by weight of peppermint extract, 1 to 3 parts by weight of parsley extract, and 0.1 to 2 parts by weight of parsley extract based on 100 parts by weight of seokchangpo extract. that includes wealth
A food for the prevention or improvement of ischemic reperfusion injury. The method of claim 1,
The extract is one selected from the group consisting of water, alcohols having 1 to 10 carbon atoms, and mixtures thereof with a solvent
A food for the prevention or improvement of ischemic reperfusion injury. delete

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