KR101645809B1 - Kimchi having function of protection of brain neuronal cells and method of preparing the same - Google Patents

Abstract

The present invention relates to a functional Kimchi for protecting brain nerve cells and a method for preparing the same, and more particularly, to a method for inhibiting neuronal cell death caused by oxidative stress by using powders and extracts of roots, roots, dandelions, dandelions, And a method for producing the functional kimchi.
The functional Kimchi for the protection of brain nerve cells of the present invention contains a mixture of doduck extract, hemp extract, bellflower extract, mixed extract of dandelion extract, dodok powder, hemp powder, bellflower powder and dandelion powder.

Description

[Technical Field] The present invention relates to a functional Kimchi for protecting brain nerve cells and a method for manufacturing the same,

The present invention relates to a functional Kimchi for protecting brain nerve cells and a method for preparing the same, and more particularly, to a method for inhibiting neuronal cell death caused by oxidative stress by using powders and extracts of roots, roots, dandelions, dandelions, And a method for producing the functional Kimchi.

Stroke, and dementia is the second most common cause of cancer and circulatory diseases (Korea Statistical Yearbook, Number of Deaths by Signs by Gender, 1996). Alzheimer's disease, Parkinson's diease, and stroke are among the most common brain diseases. These are the elderly people with brain diseases that are rapidly increasing in age and the number of elderly people suffering from these diseases is increasing. It is becoming a serious social problem.

Cerebral nerve diseases such as Alzheimer's disease, Parkinson's disease and stroke are known to play an important role in reactive oxygen species (ROS) such as free radicals. These brain diseases are the main etiologic factor of damage of nerve cell group by various reactive oxygen species produced by oxidative stress in brain cells (Smith MA, 1997. Neurochem.

In particular, brain tissue is known to be sensitive to free radical attack. This is because brain nerve cells have insufficient defense mechanisms and high concentration of long chain unsaturated fatty acids that are susceptible to oxidation, (Fe ^{2 +} , Cu ^{2 +} ) as catalysts (Olney et al. 1974. Brain Res. 77, 507-512).

Accordingly, as a part of efforts to develop drugs capable of effectively preventing or treating these neurological diseases, researches on substances exhibiting antioxidative action have been actively conducted. A number of antioxidants have been reported to exert effects in local or whole-brain ischemia animal models (Clemens, 2000. Free Radic. Biol., 28, 1526), and a-tocopherol has been associated with Alzheimer's It is known that it alleviates the rate of disease progression in sick patients. In addition, drugs that contribute to the therapeutic effect have been used because Parkinson's disease also exhibits reactive oxygen species scavenging (Ebadi et al., 1996. Prog. Neurobiol., 48, 1). However, prevention is the most important because there is currently no way to completely treat brain diseases such as Alzheimer's disease and Parkinson's disease.

Studies on medicines for preventing brain diseases from natural products with high antioxidant activity have been tried, but researches on preventing and improving brain diseases through ingestion of foods that can be easily accessed anywhere are lacking.

Japanese Laid-Open Patent Application No. 2007-0095485 discloses a food additive containing a natural product (mixed herbal extract of licorice, wheat and control) to exhibit a neuroprotective effect. However, as a natural product, dandelion, There have been no reports on functional foods that have the effect of inhibiting brain diseases using plantain.

There have been many reports on various physiological functions of dandelion, dandelion, dandelion, bellflower, and plantain. However, functional kimchi prepared by using these powders and extracts have been shown to inhibit neuronal cell death, There is no report that the ingestion of the functional kimchi contained in the composition would prevent brain diseases by protecting the brain cells from oxidative stress.

The inventors of the present invention have investigated natural materials that inhibit brain cell death by antioxidant activity in various plants. The inventors found that kimchi extract prepared with dandelion, dodok, ganoderma, And the present invention has been completed. In addition, it has been experimentally confirmed that the present invention has a lower salt content and a higher lactic acid bacterium count than a general kimchi.

Accordingly, the present invention relates to a functional kimchi for protecting brain cells capable of preventing and ameliorating brain diseases by suppressing nerve cell death due to oxidative stress using no-side effect using the corrosion which is always seen in the Korean diet, And the like.

In order to achieve the above object, the functional Kimchi for protecting brain nerve cells according to the present invention is prepared by mixing a mixed extract of doduck extract, twig extract, bellflower extract, dandelion extract, dodok powder, ganoderma powder, .

The mixed extract is characterized in that the dandelion extract: the dandelion extract: the dandelion extract: the dandelion extract is mixed at a weight ratio of 1: 1: 1: 1.

The mixed powder may further include a plantain powder.

The mixed powder is characterized by being mixed at a weight ratio of 1: 1: 1: 1: 0.1 in the composition ratio of Doduk powder: Dodong powder: Platine powder: Plantain powder: Dandelion powder.

The dandelion powder is characterized in that it is obtained by extracting 70% of a dandelion with an extracting solvent and lyophilizing it.

The kimchi is characterized by inhibiting the death of cranial nerve cells induced by oxidative stress.

The inhibition of brain neuronal cell death is characterized by inhibition of oxidative stress by antioxidant activity.

The kimchi is characterized in that it has an effect of preventing and improving degenerative brain diseases through protection of brain nerve cells.

In order to accomplish the above object, the present invention provides a method for preparing functional Kimchi for protecting brain neurons, comprising the steps of pickling Chinese cabbage in saline; 20 to 60 parts by weight of a mixed extract of 100 parts by weight of safflower in a weight ratio of 1: 1: 1: 1 of rosemary extract, rosemary extract, bellflower extract and dandelion extract, 20 to 60 parts by weight of a mixed extract of rosemary powder, , 10 to 30 parts by weight of a mixed powder in which the dandelion powder is mixed at a weight ratio of 1: 1: 1: 1: 0.1; And a step of putting the marinade sauce added with the mixed extract and the mixed powder into the pickled Chinese cabbage.

And fermenting the fermented product at 5 캜 for one week after the fermentation step.

The present invention is based on the finding that the kimchi contained in a seasoning containing dandelion, dodok, ganoderma, platycodon, plantain powder and extract has a strong antioxidative activity and, when treated with human neuroblastoma SK-N-SH under oxidative stress conditions, It was confirmed that ROS produced is strongly cleaved and the effect of inhibiting apoptotic cell death by oxidative stress is confirmed. The strong DPPH radical scavenging activity of these cells suggests that the scavenging effect of ROS in these cells will be due to the strong antioxidative effect of the present invention.

Therefore, the functional Kimchi for protecting brain cells of the present invention inhibits the death of nerve cells induced by oxidative stress with excellent antioxidative activity.

In addition, the present invention has lower salinity and a larger number of lactic acid bacteria than general kimchi, thus further enhancing the functionality of kimchi.

The present invention using natural materials such as dandelion, dandelion, marigold, bellflower, and plantain which are edible as food materials, is a cerebral disease-improving health food having no side effects and can be easily taken anytime and anywhere. Various foods , New drug development, and drug development.

FIG. 1 is a graph showing DPPH radical scavenging experiment of common kimchi extract (NKE) and kimchi extract (DKE) of the present invention. DKE had higher DPPH radical scavenging activity than NKE. Each value is expressed as mean ± standard deviation (n = 5). Other alphabets mean that there is a statistically significant difference between the groups.
2 shows the intracellular ROS scavenging activity of the common kimchi extract (NKE) and the kimchi extract (DKE) of the present invention under oxidative stress conditions. In other words, when the human neuroblastoma SK-N-SH was treated with 500 uM H ₂ O ₂ and NKE or DKE at the same time, the production of ROS was remarkably suppressed by NKE and DKE. Each value is expressed as mean ± standard deviation (n = 5). Other alphabets mean that there is a statistically significant difference between the groups.
FIG. 3 shows that neither NKE nor DKE, which is a result of the cytotoxicity of the common kimchi extract (NKE) and the kimchi extract (DKE) of the present invention, was cytotoxic at 0.1 to 2 mg / mL. Each value is expressed as mean ± standard deviation (n = 5). NS means that there is no statistically significant difference between the groups.
FIG. 4 shows the effect of the common kimchi extract (NKE) and the kimchi extract (DKE) of the present invention against the oxidative stress to inhibit neuronal cell death. That is, NKE and DKE significantly inhibited neuronal cell death by oxidative stress when 500 uM H ₂ O ₂ and NKE or DKE were simultaneously treated in human neuroblastoma SK-N-SH. At high concentrations, DKE It was more effective than NKE. Each value is expressed as mean ± standard deviation (n = 5). Other alphabets mean that there is a statistically significant difference between the groups.

Hereinafter, a functional Kimchi for protecting brain nerve cells according to a preferred embodiment of the present invention and a method for producing the same will be described in detail.

The functional Kimchi for protecting brain cells according to an embodiment of the present invention contains a mixed extract and a mixed powder. For example, the mixed extract and the mixed powder may be added to the sauce to be used for the preparation of kimchi.

In the present invention, the mixed extract means a mixture of rosemary extract, rosemary extract, bellflower extract, and dandelion extract. Preferably, the mixed extract is a mixture of Doduck extract: Dodomai extract: Dodomai extract: Dandelion extract at a weight ratio of 1: 1: 1: 1.

Each extract can be extracted in various ways. For example, an extraction solvent may be added for hot water extraction, cold-water extraction or warm-up extraction. In this case, the extraction solvent is mixed at a ratio of 5 to 50 times by weight, and the mixture is extracted at 10 to 150 ° C for 1 to 24 hours. The extraction solvent may be at least one selected from water, lower alcohols having 1 to 4 carbon atoms, polyhydric alcohols, and mixtures thereof. As the lower alcohol having 1 to 4 carbon atoms, methanol, ethanol and the like can be used. It is preferable to use a alcohol which can be edible with a lower alcohol.

As the polyhydric alcohol, butylene glycol, propylene glycol, pentylene glycol and the like can be used. Mixtures of water and lower alcohols, mixtures of water and polyhydric alcohols, mixtures of lower alcohols and polyhydric alcohols, or mixtures of water and lower alcohols and polyhydric alcohols can be used as the mixture.

In the present invention, the mixed powder is a mixture of dodok powder, god powder, bellflower powder, and dandelion powder. In this case, the mixed powder can be mixed in a weight ratio of 1: 1: 1: 0.1 of the flax powder: the flax powder: the bellflower powder: the dandelion powder.

As another example of the mixed powder, a plantain powder may be further included. In this case, the mixed powder can be mixed in a weight ratio of 1: 1: 1: 1: 0.1 in the case of dodok powder: inone powder: blooming powder: plantain powder: dandelion powder.

Each powder can be obtained by various drying methods. For example, the powder can be obtained by drying in a usual drying manner such as dodok, ganoderma, bellflower, plantain, and dandelion by natural drying or hot air drying.

Further, the extract can be obtained by extracting the extractant with an extraction solvent, concentrating it under reduced pressure, lyophilizing it, and pulverizing it. For example, the dandelion powder can be obtained by extracting 70% of a dandelion with an extracting solvent and lyophilization.

The dandelion, dandelion, dandelion, bellflower, and plantain used in the present invention are natural fruits and herbal medicines that have been used for food for a long time, and have no side effects and do not cause tolerance to patients.

Dandelion (Taraxacum officinale ) is a perennial herbaceous plant belonging to the family Asteraceae and has been used as a medicinal plant for a long time and the leaves are edible as herbs such as salad and vegetables (Oh et al. 2000. J East Asian Soc Dietary Life 10, 129-135; Lee et al. 2007. Korean J Med Crop Sci 15, 62-66).

Codonopsis lanceolata ) is a perennial herbaceous plant belonging to the lanthomaceae family and is characterized by a unique taste and aroma. It is widely distributed in the mountains of Korea and has been used for edible and medicinal purposes since ancient times (Hwang et al. 2011. J Korean Soc Food Sci Nutr 40, 798-803; Hong et al. 2006. Korean J Food Cookery Sci 22: 181-192).

Adenophora triphylla is a medicinal plant used in folk remedies for the treatment of a variety of diseases, and roots and young sprouts in the spring are edible as herbs (Ham et al. 2009. J Korean Soc Food Sci Nutr 38, 274-279 ).

Platycodon grandiflorum DC) is a perennial herb that is native to the mountain region and is used for medicinal and edible purposes in Korea. The unique bitter and flavorful flavor of the cooked food enhances the palatability of the food and has better physiological activity, making it more utilized for food than for medicinal purposes (Lee et al., 2013. J Korean Soc Food Sci Nutr 42, 1405 ~ 1411).

Plantago asiatica L. is a perennial herbaceous plant that grows wild in mountainous areas, roads and buildings around the country. In one room, tea beans are used as tea seeds and seedlings, both of which are known to have various physiological activities. In addition, plantaines have been eaten with leaves and stalks mixed with herbs or meat, and they have been tasted on kochujang (Jeong et al. 2004. J Korean Soc Food Sci Nutr 33, 1601-1605).

It has been experimentally confirmed that the kimchi of the present invention containing the mixed extract and the mixed powder described above has an effect of preventing and improving brain diseases by protecting the brain nerve cells. Namely, the effects of DPPH radical scavenging, intracellular ROS scavenging, and oxidative stress on human neuroblastoma SK-N-SH by H ₂ O ₂ , Respectively.

The kimchi of the present invention has a stronger antioxidative activity (DPPH radical scavenging activity) than general kimchi and strongly inhibits intracellular ROS when treated with human neuroblastoma SK-N-SH under oxidative stress conditions, It inhibits the death of brain nerve cells that are killed by oxidative stress. Such inhibition of brain neuronal cell death is accomplished by inhibiting oxidative stress by high antioxidant activity.

As described above, the present invention has an effect of preventing and ameliorating degenerative brain diseases through protection of brain cells.

In addition, the present invention has lower salinity and a larger number of lactic acid bacteria than general kimchi, thus further enhancing the functionality of kimchi.

The present invention is provided in the form of a food comprising a mixed extract of doduck extract, dodecahedron extract, bellflower extract, dandelion extract, mixed dodecahedron powder, ground powder, bellflower powder and mixed dandelion powder. Kimchi is taken as an example of food, but it can be provided in the form of various foods besides kimchi.

The sum of the mixed extract and the mixed powder may be 1 to 10% by weight of the total weight of the food. In the case of long-term intake for health and hygiene purposes or for health control purpose, it may be less than the above range, and since it has no problem in terms of safety, it can be used in an amount exceeding the above range It is not limited to the above range.

The term 'food' as used herein means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, Food, food additives, functional foods and beverages.

Examples of foods include various foods, beverages, gums, tea, vitamin complexes, functional foods, and the like. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirit, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, kochujang, mixed potatoes), sauces, confectionery (eg, snacks), candies, chocolate, gums, ice cream, milk products (eg, fermented milk, cheese, But are not limited to, pickled foods (various kinds of kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable beverages, beverages, fermented beverages, etc.), natural seasonings (e.g.

Furthermore, the present invention provides a flavor enhancer and a flavor enhancer which can be suitably used as flavorings such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers (cheese, chocolate etc.), pectic acid and its salts, alginic acid and its salts, Stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. These components may be used independently or in combination.

Preferably, the present invention is provided as a kimchi having a neuroprotective effect. Kimchi is easily ingested by everyone at an early age for eating.

The functional Kimchi for protecting brain cells of the present invention inhibits the death of brain nerve cells induced by oxidative stress. Herein, the death of cranial nerve cells is caused by oxidative stress, and the present invention inhibits or kills nerve cell death by inhibiting or blocking apoptosis pathway due to high antioxidant ability.

The present invention has the effect of preventing and ameliorating brain diseases, particularly degenerative brain diseases, through protection of brain cells. Degenerative brain diseases that can be prevented or ameliorated by the present invention include Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, and Hypoxic-ischemic injury.

The functional Kimchi for protecting brain cells of the present invention can be produced, for example, by the following method.

First, cabbage is pickled in brine. For example, it is soaked in 6 to 10% saline for 10 to 20 hours. Pickled Chinese cabbage and placed on a sieve for half a day to remove moisture.

Next, prepare a marinade. The sauce is made by mixing red pepper powder, garlic, ginger, onion, glutinous rice porridge, fish sauce, shrimp sauce and sugar. Add the mixed extract and the mixed powder to the prepared sauce. 20 to 60 parts by weight of the mixed extract and 10 to 30 parts by weight of the mixed powder may be added to 100 parts by weight of the sauce.

Next, the sauce prepared with the mixture of the mixed extract powder is uniformly mixed with pickled Chinese cabbage to prepare kimchi.

On the other hand, a fermentation step in which the fermentation is performed under certain conditions after the fermentation can be further performed.

For example, it is stored at 5 ° C for 1 week to ferment. The fermented kimchi thus obtained is excellent in taste, low in salinity, and large in the number of lactic acid bacteria, which is very useful for health.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following examples.

(Example)

1. Preparation of mixed extract

Dandelion, ganoderma, bellflower and dodeca were purchased from Gangwon - do, and dried and pulverized. Then, 1500 mL of drinking water was added to 50 g of each material, and the mixture was extracted at 90 ° C. for 2 hours and 30 minutes, followed by filtration to prepare dandelion extract, dendrimer extract, bellflower extract, and doduck extract. Each extract was mixed at a constant weight ratio to make a mixed extract.

2. Preparation of mixed powder

The ginseng, bellflower, doduck and plantain produced in Hoengseong, Gangwon province were purchased, dried and pulverized to prepare a powdery ginger powder, a bellflower powder, a ginger powder, and a plantain powder.

And the dandelion powder was prepared as follows.

Dandelions produced in Hoengseong, Gangwon Province were purchased and dried and ground. To 50 g of ground dandelion was added 500 mL of a 70% concentration alcohol and the mixture was extracted at room temperature (20 ° C) for 8 hours.

This procedure was repeated three times for a total of 24 hours, followed by filtration to obtain a liquid dandelion extract having a solid content removed. Then, the mixture was concentrated under reduced pressure using a vacuum rotary evaporator (Rotary Vacuum Evaporator), lyophilized using a vacuum freeze dryer, and pulverized to prepare dandelion powder.

The above-prepared powder, bellflower powder, dodok powder, plantain powder, and dandelion powder were mixed at a constant weight ratio to prepare a mixed powder.

3. Selection of optimum mixing ratio of mixed extract and mixed powder

Mixed extracts obtained by mixing the various extracts at various weight ratios were selected by nine sensory test agents to evaluate the flavor, wild taste, unique taste, and overall acceptability. The mixed extract was a mixture of Dodok extract: Dodomai extract: Dodomai extract: Dandelion extract at a weight ratio of 1: 1: 1: 1.

Mixed powders were mixed with various powders at various weight ratios and flavor, wild taste, unique taste, and overall acceptability were evaluated by 9 sensory test agents. The mixed powder having the highest score was selected as the result of the evaluation. The mixed powder was a mixture of hederdeal powder: finer powder: blooming powder: plantain powder: dandelion powder at a weight ratio of 1: 1: 1: 1: 0.1.

4. Manufacture of Kimchi

The Chinese cabbage was cut into 4 pieces and then immersed in salted 8% salt water for 15 hours to remove the cabbage. The pickled Chinese cabbage was placed on the sieve for about 6 hours to remove water.

Then, 210g of unsweetened and 20g of water were added to 125g of marinade based on 1kg of pickled cabbage, and 45g of mixed extract and 20g of mixed powder were added. The sauce was prepared by mixing 43 g of red pepper powder, 29 g of garlic, 5 g of ginger, 10 g of onion, 20 g of glutinous rice bran, 8 g of black curry sauce, 5 g of salted salted ginger, and 5 g of sugar.

The sauce mixed with water, water, mixed extract and mixed powder was mixed with pickled Chinese cabbage to prepare the kimchi (DK) of the present invention.

(Comparative Example)

In the general kimchi (NK) to be used as a comparative example, the mixed extract and the mixed powder were not added to the sauce, and more of the safflower sauce, shrimp sauce, and water were added, and the other conditions were the same.

Table 1 summarizes the Kimchi formulations of the above Examples and Comparative Examples.

material Comparative Example (NK) Example (DK) Pickled cabbage 1000g 1000g Unhealthy 210g 210g Chili powder 43g 43g garlic 29g 29g ginger 5g 5g onion 10g 10g Glutinous rice porridge 20g 20g Kannari sauce 16g 8g Shrimp 10g 5g Sugar 5g 5g water 65g 20g Mixed extract - 45g Mixed powder - 20g

<Sensory Test>

The sensory evaluation of kimchi according to the examples and the comparative examples was conducted by selecting 40 male and female college students aged 20 to 25 years using the 5 point scale method. In the sensory test questionnaires, the comparative kimchi was used as a control group and evaluated in comparison with the kimchi of the example.

The sensory evaluation items were kimchi 's appearance, color, flavor, texture, taste, salty taste, sour taste, wild taste, It is very good (5 points), good (4 points), just (3 points), strong (4 points), just (3 points), weak (2 points), very weak (1 point) Respectively.

The volunteers who participated in the sensory test recorded the age and sex in the questionnaire, and 10 g of kimchi was put into the mouth, chewed for 20 seconds to stimulate the oral skin evenly, and then spit out. When the evaluation of one sample was finished, And the following samples were evaluated.

The results of the sensory test are shown in Table 2 below.

Item Comparative Example (NK) Example (DK) Appearance 3.28 ± 0.99 3.45 ± 0.88 Color 3.35 ± 1.00 3.23 ± 0.86 Flavor 3.15 ± 0.92 2.83 ± 0.78 Texture 3.25 ± 0.67 3.48 ± 0.64 Taste 2.93 ± 0.80 2.63 ± 0.87 Salty taste 2.80 0.93 2.83 ± 0.75 Sour taste 1.35 0.48 2.45 ± 1.06 Wild grass taste 1.53 + - 0.78 3.58 ± 0.98 Comprehensive preference
(overall acceptability) 2.75 0.93 2.88 ± 0.76 Very good (5 points), good (4 points), so (3 points), bad (2 points), very bad (1 point)
Salty, sour, wild grass taste: very strong (5 points), strong (4 points), just (3 points), weak (2 points), very weak (1 point)

Referring to the results of Table 2, the appearance, color, texture, taste, and salty taste of the kimchi of the examples were similar to those of the comparative example, but the flavor was slightly lower than that of the kimchi of the comparative example. The kimchi of the example seemed to be stronger than the kimchi of the comparative example. Comprehensive preference scores were similar between the kimchi of the comparative example and the kimchi of the example.

Therefore, the kimchi of the examples and the comparative examples were similar in consumer preference, but the kimchi of the examples had excellent merits in terms of functionalities showing the effect of improving the brain disease due to the antioxidative effect, functionality containing a lot of lactic acid bacteria and low salt kimchi .

< Optimal Kimchi Selection>

Sensory evaluation was carried out using three kinds of kimchi to select the optimum kimchi according to the amount of mixed extract and mixed powder.

First, three types of kimchi were classified as DK, DK1, and DK2. DK is the kimchi of the above embodiment.

DK1 was prepared by adding 210 g of water and 20 g of water to 125 g of marinade based on 1 kg of pickled cabbage, adding 45 g of mixed extract and 10 g of mixed powder. DK2 was prepared by mixing 210 g of water and 20 g of water in 115 g of marinade based on 1 kg of pickled cabbage and adding 45 g of mixed extract. The marinades used in DK1 and DK2 are the same as the marinades used in DK.

As a result of sensory evaluation of the comparative example kimchi (NK), DK kimchi, DK1 kimchi and DK2 kimchi by the above-mentioned sensory evaluation method, DK obtained higher scores than DK1 and DK2, Respectively.

< pH , Acidity and salinity measurement>

The kimchi (DK) of the example and the kimchi (NK) of the comparative example were transferred into a plastic container, sealed, and then fermented at 5 캜 for 1 week. The pH, acidity and salinity of each kimchi were measured during fermentation. pH, acidity and salinity were determined by homogenizing 10 g of sample with 100 mL of distilled water, filtering it, measuring it with a pH meter (Pocu-ph, Sartotius, Gottingen, Germany) and acidity by AOAC standard test method (AOAC. The concentration of 0.1N NaOH consumed until the pH reached 8.4 was measured by adding 0.1N NaOH to the sample according to the Association of Official Analytical Chemistry, Washington, DC, USA, p79. The appropriate value was expressed as% of lactic acid content, and the formula was calculated as acidity (%) = [(mL of 0.1 N NaOH × normality of NaOH × 0.09) / weight of sample (g)] × 100 Respectively. Salinity was measured with a salinity meter (PAL-ES3, ATAGO, Tokyo, Japan).

As shown in Table 3 below, the pH and the pH of the comparative example and the fermentation example 3 were similar to those of 6.60 and 6.22, respectively. On the 7th day of fermentation, the pH and the pH of the control were decreased to 5.63 and 5.20, respectively. The initial pH values of the comparative example and the example were 0.13% and 0.19%, respectively, but increased during the fermentation period, and the acidity of the comparative examples and the examples were 0.39% and 0.41% at the 7th day of fermentation. On the third day of fermentation, the salinity of the comparative example and the example were 1.6% and 1.4%, respectively, and there was no change in salinity on the 7th day of fermentation. Through the salinity measurement, the samples showed lower salinity than the comparative example.

Comparative Example (NK) Example (DK) pH Fermentation 3 days 6.60 + - 0.01 6.22 ± 0.01 7 days of fermentation 5.63 ± 0.01 5.20 ± 0.01 Acidity (%) Fermentation 3 days 0.13 + - 0.01 0.19 ± 0.01 7 days of fermentation 0.39 ± 0.01 0.41 + - 0.01 Salinity (%) Fermentation 3 days 1.6 ± 0.02 1.4 ± 0.01 7 days of fermentation 1.6 ± 0.03 1.4 ± 0.01

&Lt; Measurement of number of lactic acid bacteria &

Lactic acid bacteria were prepared by mixing 10 g of Kimchi with 90 mL of sterilized water and diluting by deciduous method. Using a pour plate method using BCP agar (Eiken chemical Co. LTD., Tokyo, Japan), 37 incubators (JSGI-50L, JSR, Chung- After culturing for 24 hours, the number of colonies was counted and the number of lactic acid bacteria was measured by the number of colony forming units per gram (CFU / g).

The results of measuring the number of lactic acid bacteria in the kimchi (DK) of the examples and the kimchi (NK) of the comparative examples are shown in Table 4 below. On the third day of fermentation, the numbers of the lactic acid bacteria in the comparative examples and the examples were similar, but on the seventh day of fermentation, the number of the lactic acid bacteria in the examples was remarkably higher than those of the comparative examples.

Number of lactic acid bacteria (average, CFU / g) Comparative Example (NK) Example (DK) On the third day of fermentation 2.32 x 10 ⁵ 2.07 × 10 ⁵ On the 7th day of fermentation 6.95 × 10 ⁷ 1.29 x 10 ⁸

<Preparation of Kimchi Extract>

Kimchi extract (powdery form) was prepared from the kimchi (DK) of Example 3 after fermentation 3 days and the kimchi (NK) of Comparative Example.

After the kimchi was freeze-dried, 500 mL of a 70% alcohol solution was added to 50 g of the pulverized product, and the mixture was extracted at room temperature (20 ° C) for 8 hours. This procedure was repeated three times for a total of 24 hours, followed by filtration to obtain a liquid Kimchi extract in which the solid content was removed. The resulting kimchi extract was concentrated under reduced pressure using a rotary vacuum evaporator and then lyophilized using a vacuum freeze dryer, followed by pulverization to obtain a powdery kimchi extract. The extracted kimchi extract (DKE) extracted from the kimchi of the example and the common kimchi extract (NKE) extracted from the kimchi of the comparative example were used in all the following experiments while being stored in a freezer at -20 ° C.

<Antioxidant Effect of Kimchi Extract>

In this experiment, the antioxidative effect of the developed kimchi extract (DKE) was compared with the general kimchi extract (NKE) by measuring DPPH radical scavenging activity. Each kimchi extract was adjusted to 50 mg / mL with 40% ethanol, diluted with methanol (0.05 ~ 10 mg / mL) and used as a sample.

1.2 ml of DPPH (1,1-diphenyl-2-picryl hydrazyl) of 1.5 × 10 ^-4 M was added to 1.8 ml of the sample, reacted at 37 ° C. for 30 minutes, and the absorbance was measured at 517 nm. The radical scavenging activity was expressed by the absorbance reduction rate of the sample and the control. The control group used distilled water instead of the sample. The radical scavenging performance was calculated by the following equation and is shown in Fig.

Radical scavenging activity (%) = (1-absorbance of sample added / absorbance of control) × 100

Referring to FIG. 1, DKE scavenged DPPH radicals in a concentration-dependent manner, and showed an antioxidative effect higher than that of NKE in the group treated with 2.5-10 mg / mL.

<Kimchi extract Neuronal cell  Measurement of death suppression effect>

In this experiment, the inhibitory effect of the developed kimchi extract (DKE) on the neuronal cell death under the oxidative stress condition was compared with that of the ordinary kimchi extract (NKE).

1. Cell culture and treatment

10% inactivation of human neuroblastoma SK-N-SH in EMEM culture medium containing fetal bovine serum and penicillin / streptomycin 1% penicillin / streptomycin was performed in an incubator with 5% CO ₂ at 37 ° C Lt; / RTI &gt;

To obtain sufficient cells, cells were cultured in a culture flask (T75 cm ² ), and the medium was replaced 80% every 2-3 days. When grown sufficiently, trypsin-EDTA (ethylene diamine tetraacetic acid) solution (1x) After the adhered cells were removed, the cells were adjusted to 1 × 10 ⁴ cells / well and 2 × 10 ⁴ cells / well in the growth medium and cultured for 24 hours in 96-well plates. Were used to detect ROS.

Development Kimchi extract (NKE) and general kimchi extract (NKE) were prepared at a concentration of 50 mg / mL with 40% ethanol, diluted to a certain concentration with the medium, and used as a sample. A sample prepared by diluting the growth medium with DKE was used as a test sample, and a diluted sample was prepared by adding a growth medium to a common kimchi extract (NKE).

The cytotoxicity of kimchi extract was examined by diluting each sample with growth medium without the addition of fetal bovine serum and antibiotics for 24 hours, and the results are shown in FIG.

In order to examine the protective effect of the Kimchi extract against oxidative stress, each sample was simultaneously treated with 500 μM H ₂ O ₂ for 30 minutes. The results are shown in FIG. In FIG. 4, the control group (C) and the positive control group (H ₂ O ₂ ) are the untreated group and the group treated with 500 μM H ₂ O ₂ , respectively.

2. Measurement of reactive oxygen species (ROS) in cells

SK-N-SH cells were pretreated with 2,7-dichlorofluorescein diacetate (DCF-DA) as a prove to determine whether oxidative stress was present and whether the samples could eradicate ROS in brain cells. The amount of ROS produced in N-SH cells was measured. 10 mM DCF-DA was made and diluted with PBS to a concentration of 10. After the medium was removed from the cells, 10DCF-DA solution was added to the cells, followed by incubation at 37 ° C for 45 minutes. After removal of DCF-DA solution, DCF-DA remaining in PBS was completely removed, and PBS was added and incubated for 5 minutes. Changes in intracellular fluorescence intensities were measured at 488 nm (excitation) and 525 nm (emission) using a fluorescence photometer.

The test sample and the comparative sample, respectively are shown in Figure 2, the results when treated simultaneously with 500μM H ₂ O _2. In FIG. 2, the control group (C) and the positive control group (H ₂ O ₂ ) are the untreated group and the group treated with 500 μM H ₂ O ₂ , respectively.

Referring to FIG. 2, DKE and NKE treatment significantly inhibited increased intracellular ROS by 500 μM H ₂ O ₂ . The inhibitory effect of DKE on intracellular ROS production was concentration-dependent, and DKE had a higher ROS scavenging activity than the NKE (0.5 mg / mL, 1.0 mg / mL), but no statistically significant difference was observed.

3. Measurement of cell viability

The survival rate of cultured SK-N-SH cells was measured by the MTT [3- (4,5-dimethythiazol-2-yl) -2,5-dipheyl terazolium bromide] reduction method used by Chung et al. That is, each well MTT solution (5 mg / mL) was added to 1/10 of the growth medium and further cultured at 37 ° C. for 4 hours to reduce the MTT. The medium was carefully removed so that the generated formazon did not follow the medium . To completely remove the remaining medium, the sample was allowed to stand at room temperature for 30 minutes and then dissolved in DMSO. Absorbance was measured at 570 nm. For the measurement of absorbance, the control group was DMSO and the cell viability was calculated as follows.

Cell survival rate (%) = [absorbance of sample treated group / absorbance of control group] x 100

Referring to FIG. 3, no cytotoxicity was observed when NKE and DKE at a concentration of 0.1 to 2 mg / mL were treated alone.

And, Figure 4, compared with the general kimchi extract (NKE) or development kimchi extract when (DKE) the process at the same time with _{500μM H 2 O 2 500μM H 2} O 2 only treatment of the positive control (H ₂ O ₂₎ The cell survival rate of DKE treated group was significantly higher than that of NKE treatment group at 0.25 ~ 1mg / mL concentration.

As shown in the above experiment, it was confirmed that the functional kimchi of the present invention containing natural materials more effectively protects brain cells against oxidative stress than general kimchi. These results fully support that the present invention is a health-functional kimchi capable of preventing or treating various degenerative brain diseases including Alzheimer caused by oxidative stress.

<Statistics Processing>

The test results were expressed as mean ± SD, and the mean difference between the experimental groups was analyzed by one-way ANOVA using Duncan's multiple range test. P <0.05 The significance was tested. All statistical analyzes were performed using the Statistical Package for the Social Science (SPSS) version 12.0 program (SPSS Inc., Chicago, IL, USA).

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation. Accordingly, the true scope of protection of the present invention should be determined only by the appended claims.

Claims (10)

A pickling step in which the cabbage is soaked in 6 to 10% saline for 10 to 20 hours;
20 to 60 parts by weight of a mixed extract of 100 parts by weight of safflower in a weight ratio of 1: 1: 1: 1 of rosemary extract, rosemary extract, bellflower extract and dandelion extract, 20 to 60 parts by weight of a mixed extract of rosemary powder, , 10 to 30 parts by weight of a mixed powder in which the dandelion powder is mixed at a weight ratio of 1: 1: 1: 1: 0.1;
A step of putting the marinade sauce added with the mixed extract and the mixed powder into the pickled Chinese cabbage;
A fermentation step of fermenting at 5 캜 for one week following the step of fermentation;
Wherein the method comprises the steps of: delete delete delete The method according to claim 1,
Wherein the dandelion powder is obtained by extracting 70% of a dandelion with 70% alcohol as an extraction solvent, followed by lyophilization, to obtain a functional kimchi. The method according to claim 1,
Wherein the kimchi inhibits the death of brain nerve cells induced by oxidative stress. The method according to claim 6,
Wherein said inhibition of brain neuronal cell death is caused by inhibition of oxidative stress induced by antioxidant activity. The method according to claim 1,
Wherein the kimchi has an effect of preventing and ameliorating degenerative brain diseases through protection of brain nerve cells. delete delete

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