KR20000000375A - Anticancer activities of chinese cabbage Kimchi and process for preparation thereof - Google Patents

Abstract

PURPOSE: A cabbage Kimchi of an anti-cancer function and a production method thereof are provided to show the anti-cancer function not only in the in vitro but also in the in vivo, and that is excellent in the preservation. CONSTITUTION: A cabbage Kimchi of the anti-cancer function comprises: radish 13.0%; leek 2.0%; powder of red pepper 7%; garlic 2.8%; ginger 0.6%; salted anchovies 2.2%; sugar 1.0%; refined salt 2.5%; leaf mustard 5% mixed in salted cabbage 100; fermented the first in 15°C; re-fermented in a low temperature of 5°C to pH4.3.

Description

Anti-cancer functional cabbage kimchi and its manufacturing method {Anticancer activities of chinese cabbage Kimchi and process for preparation}

The present invention relates to an anti-cancer functional cabbage kimchi and a method of manufacturing the same. More specifically, in the production of cabbage kimchi, the present invention relates to anti-cancer functional cabbage kimchi and anti-cancer cabbage kimchi which improved anticancer functionality of cabbage kimchi by adjusting the mixing ratio of materials such as red pepper powder, garlic, freshly chopped and chopi.

In modern times, it has been found that each food has a function of promoting health, and researches on each food ingredient supply, processing, and fermentation technology are actively conducted. In particular, kimchi is a food that does not fall out of our diet has been studied a lot. For example, the study on the production of kimchi with the addition of subsidiary materials is carried out by the method of ripening and extending the storage period of kimchi by the addition of chitosan oligosaccharide (Korean Patent No. 98-019544), kimchi seasoning composition containing chitin and roasted salt -13238), ginseng added cabbage kimchi (Korean Patent Publication 96-13237), ginseng kimchi manufacturing method (Republic of Korea Patent Publication 98-025327), seaweed kimchi production method (Republic of Korea Patent Publication 97-58579), mushroom kimchi The manufacturing method (Korean Patent Publication No. 97-032465) and the like are known, but these known techniques are related to the extension of the shelf life and a small amount of the additives known to be functional.

Kimchi is a fermented food Kimchi's fermentation technology has traditionally been handed down by housewives, but recently, industrialization has promoted the use of factory kimchi. With such industrialization, Kimchi's diversification and functionality are emphasized, but until now, Kimchi, which has anticancer function, has been mainly studied to obtain the effect by simply adding ingredients with anticancer effect to kimchi. No research has been published on the production of anti-cancer functional kimchi by controlling the amount of the ingredients.

In order to solve the above problems, the present inventors have prepared various kinds of cabbage kimchis according to the kinds of ingredients, their mixing ratios and fermentation conditions, and then investigated the anticancer functionality of these cabbage kimchis in vitro as well as in vivo. The present invention was completed by examining the mixing ratio and fermentation conditions of the cabbage kimchi that is effective.

Therefore, an object of the present invention is to provide a cabbage kimchi exhibiting anti-cancer functionality by mixing the standardized cabbage kimchi in an appropriate ratio of the subsidiary materials and fermented under specific conditions. Another object of the present invention to provide a method for producing the anti-cancer functional cabbage kimchi. Another object of the present invention to provide a diet containing the anti-cancer functional cabbage kimchi.

The purpose of the present invention is to make kimchi and organic cabbage kimchi dipped by varying the amount of red pepper powder, garlic, fresh, auditory, cinnamon, cinnamon powder, ginseng powder, chitosan oligosaccharides, alginic acid, additive powder based on standardized Chinese cabbage kimchi pH 4.3 The anticancer and antimutagenic effects of Chinese cabbage kimchi were fermented in various ways until fermentation, and the anticancer effect of methanol extract from fermented cabbage kimchi was investigated in vitro. Subsequently, using the organically grown ingredients and preparing cabbage kimchi with the addition of red pepper powder, garlic, freshly and chopi to the standardized kimchi's ingredients, the cancer prevention and anticancer effects of the cabbage kimchi were also investigated in vitro. The anticancer and immune activity enhancement effects of Chinese cabbage kimchi prepared with the organically grown material in vivo and the effects on nitric oxide production of mouse macrophages were investigated. Furthermore, after feeding a diet containing an appropriate amount of cabbage kimchi made of the organic material to the mice, the anti-cancer effect and immune-enhancing effects were also investigated, and after the subcutaneous injection of methanol extract of the organic cabbage kimchi into the mouse, the metastasis to the lungs was increased. A tumor cell line was injected into the tail of the mouse and then examined by examining tumor suppression effects.

Hereinafter, the configuration and operation of the present invention.

1 shows nitric oxide (NO) measured by the concentration of nitrate ion generated in intraperitoneal macrophages of mice treated with cabbage kimchi extract of the present invention.

Figure 2 shows the effect of the anticancer functional cabbage kimchi methanol extract of the present invention on glutathione-S-transferase (GST) activity in mouse liver treated with sarcoma-180 cells.

Figure 3 shows the effect of the anticancer functional cabbage kimchi methanol extract of the present invention on glutathione (GSH) content in mouse liver treated with sarcoma-180 cells.

Figure 4 shows the effect of the anticancer functional cabbage kimchi methanol extract of the present invention on the content of lipid peroxide in mouse liver treated with sarcoma-180 cells.

5 shows the effect of the anticancer functional cabbage kimchi methanol extract of the present invention on the natural killer cell activity of mouse spleen lymphocytes.

Figure 6 shows the effect of the present cabbage kimchi containing diet on the natural killer cell activity of mouse spleen lymphocytes.

Figure 7 is a graph showing the anti-nuclear induction effect of the anti-cancer functional cabbage kimchi of the present invention compared to the standardized cabbage kimchi.

13 kinds of Chinese cabbage kimchi with different amounts of supplementary ingredients such as roasted salt, red pepper powder, garlic, lampshade, auditory, cinnamon, cinnamon, ginseng, chitosan oligosaccharide, sodium alginate, and powdered powder, which have anti-cancer functional effects Manufacturing step; Culturing AGS human adenocarcinoma cells, HT-29 human colon adenocarcinoma cells, and MG-63 human osteosarcoma cells for use in anticancer effect experiments; Comparing the time required for reaching 13 kinds of Chinese cabbage kimchi prepared by varying the amount and type of subsidiary ingredients to standardized Chinese cabbage kimchi at different fermentation temperatures to pH 4.3, which is the final ripening season of kimchi (comparable to preservation); 13 kinds of Chinese cabbage kimchi prepared by varying the amount and type of subsidiary ingredients in standardized Chinese cabbage kimchi were fermented at 15 ° C. for 1 day and then fermented again until pH 4.3 at 5 ° C. Comparing and analyzing the survival inhibition effect on the cells by MTT assay; The survival inhibition effect of the cabbage kimchi juice fermented from 15 ℃ to pH 4.3 at 15 ℃ was prepared by MTT assay. Comparative analysis; The survival inhibition effect of Chinese cabbage kimchi juice obtained by fermenting 13 kinds of Chinese cabbage kimchi prepared by varying the amount and type of subsidiary materials to standardized cabbage kimchi until pH 4.3 at 25 ℃ was investigated by MTT assay. Comparative analysis; Investigating the antimutagenic effect of methanol extracts from 13 kinds of Chinese cabbage kimchi prepared by varying the amount and type of subsidiary ingredients in the standardized Chinese cabbage kimchi by Ames test using Salmonella typhimurium TA100 strain; Confirming the antimutagenic effect of methanol extracts obtained from 13 kinds of cabbage kimchi prepared by varying the amount and type of subsidiary ingredients to standardized Chinese cabbage kimchi by SOS chromotest; AGS Human Gastric Cancer Cells, HT-29 Human Colon Cancer Cells, HL-60 Human Blood Cancer Cells, and MG-63 Human Osteosarcoma Cells of Methanol Extracts from 13 Chinese Cabbage Kimchi Prepared with Different Substances and Types of Standardized Chinese Cabbage Kimchi Comparing with a sulforhodamine B assay to investigate the effect of inhibiting survival; AGS Human Gastric Cancer Cells, HT-29 Human Colon Cancer Cells, HL-60 Human Blood Cancer Cells, and MG-63 Human Osteosarcoma Cells of Methanol Extracts from 13 Chinese Cabbage Kimchi Prepared with Different Substances and Types of Standardized Chinese Cabbage Kimchi Investigating survival inhibition by MTT assay; Attaching AGS human gastric cancer cells and HT-29 human colon cancer cells to the plate and adding fermented cabbage kimchi methanol extract as a sample and culturing the cells to investigate the effect of inhibiting cancer cell growth by measuring the number of proliferated cells; Standardized Chinese Cabbage Kimchi, Organic Standardized Chinese Cabbage Kimchi Using Organically Grown Ingredients, General High-Active Chinese Cabbage Kimchi Using Organic Ingredients and Added Red Pepper Powder, Garlic, Fresh, and Chopi Preparing organic high-activity Chinese cabbage kimchi added with red pepper powder, garlic, freshly and chopi to the Chinese cabbage, fermenting and lyophilizing, and then extracting methanol to obtain an extract; Investigating oral administration of methanol extracts of the standardized cabbage kimchi, organic standardized cabbage kimchi, general high activity cabbage kimchi, and organic high activity cabbage kimchi to mice and observing the effect of micronucleus suppression to investigate the cancer prevention effect; Investigating the inhibitory effect of AGS human gastric cancer cells of the methanol extract obtained from the standardized cabbage kimchi, organic standardized cabbage kimchi, general high activity cabbage kimchi, organic high activity cabbage kimchi by MTT assay; performing a cytotoxicity test of the methanol extract of the standardized cabbage kimchi, organic standardized cabbage kimchi, general high activity cabbage kimchi, and organic high activity cabbage kimchi by performing a survival test in vitro using a dye exclusion method; Implantation of 1.0 × 10 ⁶ cell / mL tumor cell suspension in the abdominal cavity of the male Balb / c mouse and the standardized cabbage kimchi, organic standardized cabbage kimchi, general high activity cabbage kimchi, organic high activity cabbage Administering a sample of methanol extract obtained from kimchi and observing survival over time to observe the life extension effect and to investigate the weight change of each organ; After subcutaneous transplantation of tumor cell suspension into mice, the standardized Chinese cabbage kimchi, organic standardized Chinese cabbage kimchi, general high activity cabbage kimchi, and methanol extract samples of organic high activity cabbage kimchi were administered to the abdominal cavity, and the resulting solid cancer was extracted and weighed. Measuring; Glutathione-S-transferase (GST) activity and glutathione in the mouse liver administered with the methanol extract of the standardized Chinese cabbage kimchi, organic standardized Chinese cabbage kimchi, general high active cabbage kimchi, organic high active cabbage kimchi Examining the change in the content of GSH; NK cells were isolated from tumor cells and spleens of mice to which the standardized Chinese cabbage kimchi, organic standardized Chinese cabbage kimchi, general high activity cabbage kimchi and methanol extract of organic high activity cabbage kimchi were isolated, and their cell destruction on Yac-1 cells. Measuring activity (% target cell cytolysis); After administering the standardized Chinese cabbage kimchi, organic standardized Chinese cabbage kimchi, general high activity cabbage kimchi, organic high activity cabbage kimchi methanol extract for 3 weeks into the mouth of the mouse, the intraperitoneal macrophages are separated to act by lipopolysaccharide (lipopolysaccharide) Measuring the amount of nitric oxide (NO) produced; Injecting sarcoma-180 cells into the inguinal part of the mice fed the diet containing the standardized Chinese cabbage kimchi, organic standardized Chinese cabbage kimchi, general high-active cabbage kimchi, organic high-active cabbage kimchi and continued feeding the diet to solid cancer growth inhibition And investigating the activity of natural killer cells; Inhibition of Chinese cabbage kimchi extract subcutaneously, two days later, intravenously injected with colon 26-M3.1 carcinoma, a classical tumor cell line, in the tail of Balb / c mice, and 14 days later, lung tumors were removed to inhibit tumor metastasis. Investigate the effect.

In the present invention, the standardized Chinese cabbage kimchi of the Department of Food and Nutrition, Pusan National University, Eun-ju Cho, Sun-mi Lee, Sook-hee Lee, and Kun-young Park were standardized cabbage kimchi recipes through sensory and physicochemical tests according to the mixing ratio of Chinese cabbage kimchi. The ideal cabbage kimchi composition ratio, published in the Korean Journal of HSKCAN 39 (2), is an ideal cabbage kimchi blending ratio for pickled cabbage 100 radish 13.0, leek 2.0, red pepper powder 3.5, garlic 1.4, ginger 0.6, salted anchovy 2.2, sugar 1.0 Cabbage kimchi containing 2.5 final salt.

Human gastric cancer cells (AGS human adenocarcinoma cell), HT-29 human colon adenocarcinoma cell, HL-60 human promyelocytic leukemia cell, MG-63 human bone Cancer cells (MG-63 human osteosarcoma cells) were distributed from the Korea Cell Line Bank (Seoul National University of Medicine), and sarcoma 180 cells from mice are being stored in the laboratory. Colon 26-M3.1 carcinoma, a classical tumor cell line used, was purchased from Dr. Yoon Taek-joon of Handong University and the animal Balb / c mouse (female, 6 weeks old) was purchased from the Korea Research Institute of Chemical Technology in Daejeon.

Salt samples used in the present invention are two kinds of purified mechanical salts commercially available (NaCl reagent (Junsei Chemical Co., Ltd) and Hanjugeum)] and two kinds of natural salts [west coast sun salts purchased from Sansan Co., Ltd.] , Raw salt Co., Ltd.] and three kinds of processed salts (baked salt Co., Ltd.). Mountain salts, bamboo salt (hazelnut) were used, respectively.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1: 13 kinds of Chinese cabbage kimchi

Chinese cabbage was pickled in brine (10%) twice the weight of cabbage for 10 hours, washed three times, and then used to remove water for 2 hours. Based on the standardized Chinese cabbage kimchi, 13 kinds of Chinese cabbage kimchi were prepared by different kinds and amounts of subsidiary materials. Standardized Chinese cabbage kimchi, Kimchi with 10.5% red pepper powder, Kimchi with 5.2% garlic, Kimchi with 7% red pepper powder + 2.8% Garlic, Kimchi with 5% fresh garlic, Kimchi with 5% fresh hearing, Kimchi with 1% cinnamon powder, Kimchi with 1% cinnamon powder , Kimchi with 1% ginseng powder, kimchi with 1% chitosan oligosaccharide, kimchi with 1% sodium alginate, kimchi with 1% added powder, kimchi with organic cabbage instead of regular cabbage The temperature treatment was changed to pH 4.3 or fermented at 15 ° C. for 1 day and fermented at 5 ° C. to obtain a juice solution and a methanol extract. At this time, the salt used in this embodiment may be any one of reagent NaCl, mechanical salt, sun salt, fresh salt, baked salt, fresh salt, bamboo salt, but the growth inhibitory effect of HT-29 human colon cancer cells is the best and DNA of cancer cells Roasted salt with the best synthetic inhibitory effect was used.

Baked salt was treated in HT-Human Colon Cancer cells and tested for its growth inhibition effect. As shown in Table 1, NaCl for reagent, 19% for Meth salt, 33% for Natural Salt, 31% for Raw Salt, and 60% for Baked Salt. Inhibition of 57% and 50% of bamboo salt showed the highest effect. In addition, as shown in Table 2, the inhibition of DNA synthesis using HT-29 cancer cells showed that NaCl was 28%, mechanical salt 40%, sun salt 45%, fresh salt 30%, baked salt 82%, and salt at 10 μl / well. Inhibition of DNA synthesis by 83% and bamboo salt by 61% was also observed.

Inhibitory Effect of Salt on HT-29 Human Colon Cancer Cell Growth Concentration per well 1 µg 5 µg 10 µg 20 µg NaCl for reagents 32 ± 3 (24) ¹⁾ 31 ± 1 (23) 36 ± 7 (14) 35 ± 3 (17) Mechanical salt 34 ± 6 (19) 34 ± 6 (19) 30 ± 6 (29) 40 ± 7 (5) Sun salt 35 ± 1 (17) 28 ± 4 (33) 26 ± 1 (38) 30 ± 6 (29) Fresh salt 34 ± 4 (19) 29 ± 11 (31) 30 ± 1 (29) 30 ± 4 (29) Baked Salt 32 ± 5 (24) 17 ± 3 (60) 18 ± 5 (57) 12 ± 7 (71) Salary 20 ± 0 (52) 18 ± 7 (57) 15 ± 7 (64) 13 ± 5 (69) Bamboo salt 24 ± 2 (43) 21 ± 8 (50) 21 ± 8 (50) 21 ± 12 (50) [Note] Unit: number of cells × 10 ⁴ / mL (1): inhibition rate

Inhibitory Effects of Salt on HT-29 Human Colon Cancer Cell DNA Synthesis Concentration per well 1 µg 5 µg 10 µg 20 µg NaCl for reagents 1.65 ± 0.06 (20) ¹⁾ 1.80 ± 0.09 (12) 1.48 ± 0.44 (28) 1.86 ± 0.33 (10) Mechanical salt 2.00 ± 0.67 (3) 1.82 ± 0.29 (12) 1.22 ± 0.17 (40) 2.17 ± 0.04 (-5) Sun salt 2.03 ± 0.96 (1) 0.89 ± 0.12 (8) 1.13 ± 0.23 (45) 1.82 ± 0.02 (12) Fresh salt 1.91 ± 0.17 (7) 1.81 ± 0.17 (12) 1.44 ± 0.23 (30) 1.55 ± 0.31 (25) Baked Salt 2.08 ± 0.51 (1) 1.40 ± 0.19 (32) 0.37 ± 0.08 (82) 0.42 ± 0.02 (79) Salary 1.92 ± 0.68 (7) 1.38 ± 0.05 (33) 0.35 ± 0.10 (83) 0.30 ± 0.01 (85) Bamboo salt 1.65 ± 0.75 (20) 1.31 ± 0.20 (36) 0.81 ± 0.14 (61) 0.66 ± 0.30 (68) [Note] (1) Inhibition rate

Example 2: Cancer Cell Culture

AGS human gastric cancer cells and HT-29 human colon cancer cells were cultured in a 37 ° C., 5% CO ₂ incubator using RPMI 1640 containing 100 units / mL penicillin-streptomycin and 10% FCS. Each cultured cancer cell was refeeded 2-3 times a week with fresh medium, washed with PBS in 6-7 days, and then centrifuged by separating cells attached with 0.05% trypsin-0.02% EDTA. The medium was added to the integrated cancer cells, and the pipettes were mixed well so that the cancer cells were evenly dispersed. The cells were inoculated by a predetermined number of 10mL portions in a 75mL cell flask, and the cells were continuously cultured every 6 to 7 days. Each passage number was recorded at the time of passage and when more than 10 passages were taken, new cancer cells were removed from the liquid nitrogen tank and tested again.

Example 3: In vitro anticancer effect of Chinese cabbage kimchi according to fermentation temperature and cabbage kimchi juice

Experimental Example 1: Experiment of storage capacity of Chinese cabbage kimchi according to fermentation temperature

In this experiment, the fermentation temperature was compared first until the pH of 4.3 increased the retention of additives. As shown in Table 3, kimchi added three times (10.5%) of red pepper powder compared to standardized Chinese cabbage kimchi, took longer to reach pH 4.3 than fermentation.

When fermentation was carried out at 15 ° C. for 1 day and fermentation at 5 ° C., the time required to reach pH 4.3 was 15 days compared to 15 days of standardized Chinese cabbage kimchi. Also, kimchi added with garlic 3 times (5.2%) was found to be more slowly fermented than standardized kimchi. Kimchi added twice with red pepper powder and garlic did not show much difference from the standardized cabbage kimchi. , Fermentation was delayed when fermented at 25 ℃.

Kimchi added 5% of fresh and auditory to Kimchi cabbage was not different from the standardized kimchi, and fermentation was delayed slightly when fermented at 25 ℃. Fermentation of kimchi with chopi, chitosan oligosaccharide, and added powder was slower than standardized cabbage kimchi. Especially, it took time to reach pH 4.3 when fermented with chitosan oligosaccharide and added powder from 15 ℃ → 5 ℃. These were 20 days and 24 days, respectively, much longer than the standardized Kimchi 15 days.

In the case of kimchi dipped with organic cabbage, fermentation occurred a little slower than standardized kimchi, but there was no significant difference.

Number of days of fermentation until days when the additive-free kimchi and organic cabbage kimchi reached pH 4.3 Fermentation Temperature Kimchi Type 15 ℃ → 5 ℃ ¹⁾ 15 ℃ 25 ℃ Standardized Chinese Cabbage Kimchi 10.5% Red Pepper Powder Kimchi 5.2% Garlic Kimchi 7% Red Pepper Powder + 2.8% Garlic Kimchi 5% Fresh Kimchi 5% Hearing Kimchi 1% Chopi 1 Kimchi 1% Cinnamon Powder 1 Kimchi 1% Ginseng Powder 1 % Chitosan oligosaccharide added kimchi 1% sodium alginate added kimchi 1% added powder Kimchi organic cabbage kimchi 15181615151516201320152416 4544445446475 11.51.52.52122.512.5131

The reason for fermentation at 5 ℃ for one day at 15 ℃ is to improve the taste and fermentation period by increasing the number of lactic acid bacteria of Leuconostoc sp.

Experimental Example 2: anticancer effect of cabbage kimchi juice fermented at 5 ℃

Cancer cells were seeded at 1 × 10 ³ cells / mL per well in a 96-well plate, incubated for 24 hours, and then fermented with cabbage kimchi prepared in Example 1 before each fermentation of cabbage kimchi as a sample. C was incubated in a 5% CO ₂ incubator. After 3 days of culture, 50 µl of 3- (4,5-dimethyl-thiazol) -2,5-diphenyl tetrazolium bromide; MTT, 2 mg / mL) was added, followed by further incubation for 4 hours, and then the medium was removed (180 µl). 150 μl of DMSO was added and mixed with a plate mixer, and the absorbance was measured at 540 nm. Based on the average cell growth rate (%) obtained by treating each sample by concentration, the concentration (IC ₅₀ ) of each sample that inhibited cell growth by 50% was calculated according to the Lotus data regression program. The cabbage kimchi was fermented at 15 ° C. for 1 day and then fermented at 5 ° C. until pH 4.3 (13-20 days), and the juice was used as a sample. As a result, as shown in Table 4, 13 kinds of Chinese cabbage kimchi were fermented at 15 ° C. for 1 day and then fermented until the pH was 4.3 at 5 ° C .. It was found that kimchi and organic cabbage kimchi added 7% and 2.8%, respectively, significantly inhibited the survival of AGS human gastric cancer cells compared to standardized cabbage kimchi. Especially, kimchi added with chopi showed a high inhibitory effect. (p <0.05). Table 5 shows the inhibitory effect on the survival of HT-29 colon cancer cells of cabbage kimchi juice fermented at 15 ° C. to pH 4.3 after fermentation at 15 ° C. for 1 day. Compared to Chinese cabbage kimchi, freshly added kimchi and organic cabbage kimchi showed a high inhibitory effect (p <0.05). At 10 μl / assay juice, standardized cabbage kimchi inhibited the survival of 42% of human colon cancer cells, while freshly added kimchi and organic cabbage kimchi inhibited the survival of colon cancer cells by more than 50%. (P <0.05).

The effect of inhibiting the survival of AGS human gastric cancer cells by MTT test of the juice obtained by fermenting 13 kinds of Chinese cabbage kimchi at 15 ℃ for 1 day and then fermenting at 5 ℃ to pH 4.3 Treated substance (μl / assay) OD ₅₄₀ 5 10 Control 0.790 ± 0.05 ^a 0.618 ± 0.032 ^a Standardized Chinese Cabbage Kimchi 0.652 ± 0.012 ^bc (17) ¹⁾ 0.190 ± 0.012 ^cd (69) Kimchi with 10.5% red pepper powder 0.368 ± 0.022 ^e (53) 0.187 ± 0.022 ^cd (70) Kimchi with 5.2% garlic 0.506 ± 0.078 ^d (36) 0.233 ± 0.078 ^bcd (62) Kimchi with 7% red pepper powder + 2.8% garlic 0.768 ± 0.021 ^ab (3) 0.150 ± 0.021 ^cd (76) 5% freshly added kimchi 0.487 ± 0.088 ^de (38) 0.077 ± 0.088 ^d (88) 5% Hearing Added Kimchi 0.808 ± 0.015 ^a (-2) 0.265 ± 0.015 ^bc (57) Kimchi with 1% Chopi 0.541 ± 0.120 ^cd (32) 0.402 ± 0.012 ^b (35) Kimchi with 1% Cinnamon Powder 0.591 ± 0.087 ^cd (25) 0.166 ± 0.087 ^cd (73) Kimchi with 1% Ginseng Powder 0.652 ± 0.109 ^bc (17) 0.250 ± 0.109 ^bc (60) Kimchi with 1% chitosan oligosaccharide 0.648 ± 0.065 ^bc (18) 0.375 ± 0.065P (39) Kimchi with 1% Sodium Alginate 0.672 ± 0.069 ^abc (15) 0.371 ± 0.069 ^b (40) Kimchi with 1% added powder 0.750 ± 0.231 ^ab (5) 0.569 ± 0.231 ^a (8) Organic Cabbage Kimchi 0.603 ± 0.123 ^cd (24) 0.125 ± 0.123 ^cd (80) One) a to d represent significant differences (p <0.05) in Duncan's multiple range test.

The effect of inhibiting the survival of HT-29 human colon cancer cells by MTT test of the juice of 13 cabbage kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 Treated substance (μl / assay) OD ₅₄₀ 5 10 Control 0.703 ± 0.02 ^a 0.913 ± 0.028 ^a Standardized Chinese Cabbage Kimchi 0.483 ± 0.014 ^bc (31) ²⁾ 0.526 ± 0.034 ^bc (42) Kimchi with 10.5% red pepper powder 0.337 ± 0.072 ^cd (52) 0.662 ± 0.083 ^b (28) Kimchi with 5.2% garlic 0.510 ± 0.179 ^b (28) 0.614 ± 0.143 ^bc (33) Kimchi with 7% red pepper powder + 2.8% garlic 0.521 ± 0.063 ^b (30) 0.617 ± 0.138 ^bc (32) 5% freshly added kimchi 0.229 ± 0.101 ^d (67) 0.415 ± 0.072 ^c (55) 5% Hearing Added Kimchi 0.667 ± 0.059 ^a (5) 0.648 ± 0.078 ^b (29) Kimchi with 1% Chopi 0.382 ± 0.027 ^bc (46) 0.545 ± 0.022 ^bc (40) Kimchi with 1% Cinnamon Powder 0.453 ± 0.062 ^bc (36) 0.416 ± 0.116 ^c (54) Kimchi with 1% Ginseng Powder 0.431 ± 0.020 ^bc (39) 0.670 ± 0.266 ^b (27) Kimchi with 1% chitosan oligosaccharide 0.477 ± 0.134 ^bc (32) 0.532 ± 0.061 ^bc (42) Kimchi with 1% Sodium Alginate 0.269 ± 0.065 ^bc (40) 0.469 ± 0.015 ^bc (49) Kimchi with 1% added powder 0.481 ± 0.800 ^bc (32) 0.562 ± 0.085 ^bc (38) Organic Cabbage Kimchi 0.400 ± 0.420 ^bc (43) 0.445 ± 0.005 ^bc (51) One) a to d represent significant differences (p <0.05) in Duncan's multiple range test.

Experimental Example 3: anticancer effect of Chinese cabbage kimchi juice fermented at 15 ℃

The anticancer effect was investigated in the same manner as in Experimental Example 1, but the cabbage kimchi was fermented at 15 ° C. until pH 4.3, and the juice was used as a sample. Experimental results, as shown in Table 6 showed a somewhat lower inhibitory effect compared to the cabbage kimchi juice fermented for 1 day at 15 ℃ of Experimental Example 2 and 5 ℃. Compared to the standardized Chinese cabbage kimchi juice, kimchi containing red pepper powder three times, kimchi with red pepper powder and garlic added twice, auditory kimchi, chopi added kimchi and organic cabbage kimchi showed higher inhibitory effect on AGS human gastric cancer cell survival. p <0.05). In particular, kimchi (7%, 2.8%) with 2 times the red pepper powder and garlic respectively showed the inhibition of survival of 57% and 68% of human gastric cancer cells at 5µl / assay and 10µl / assay juice concentrations. The inhibitory effect was much higher than 40% and 59%. As a result, the effect on the survival of cancer cells was somewhat different according to the fermentation temperature. However, it was found that kimchi, chopi added kimchi and organic cabbage kimchi with high red pepper powder and garlic had higher cancer cell survival inhibition effect than standardized cabbage kimchi. Could. Table 7 also compares the effects of 13 kinds of kimchi juice fermented to pH 4.3 at 15 ° C. on the survival of HT-29 human colon cancer cells. The juices of kimchi added red pepper powder three times, kimchi added 5% freshly, fresh kimchi added 1% chopi and cabbage kimchi added 1% chitosan oligosaccharide compared to standardized Chinese cabbage kimchi, and the survival of HT-29 human colon cancer cells Greater inhibition (p <0.05). In the case of cabbage kimchi juice fermented at 15 ℃, it was found that kimchi and organic cabbage kimchi added with red pepper powder and garlic at high concentrations inhibited the survival of HT-29 human colon cancer cells as well as AGS human stomach cancer cells. there was.

Inhibition of AGS Human Gastric Cancer Cell Survival by MTT Test Method of Juice Extracted by Fermenting 13 Cabbage Kimchis to pH 4.3 at 15 ℃ Treated substance (μl / assay) OD ₅₄₀ 5 10 Control 0.795 ± 0.037 ^a 0.795 ± 0.037 ^a Standardized Chinese Cabbage Kimchi 0.476 ± 0.032 ^cd (40) ¹⁾ 0.323 ± 0.011 ^efg (59) Kimchi with 10.5% red pepper powder 0.345 ± 0.050 ^e (57) 0.256 ± 0.007 ^gh (68) Kimchi with 5.2% garlic 0.431 ± 0.095 ^de (46) 0.348 ± 0.015 ^def (56) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.389 ± 0.042 ^de (51) 0.257 ± 0.028 ^gh (68) 5% freshly added kimchi 0.565 ± 0.071 ^c (29) 0.362 ± 0.023 ^cde (55) 5% Hearing Added Kimchi 0.413 ± 0.016 ^f (48) 0.254 ± 0.031 ^gh (68) Kimchi with 1% Chopi 0.475 ± 0.050 ^cd (40) 0.298 ± 0.020 ^efgh (63) Kimchi with 1% Cinnamon Powder 0.676 ± 0.106 ^b (15) 0.234 ± 0.018 ^h (71) Kimchi with 1% Ginseng Powder 0.731 ± 0.018 ^ab (8) 0.436 ± 0.042 ^b (45) Kimchi with 1% chitosan oligosaccharide 0.484 ± 0.017 ^cd (39) 0.283 ± 0.027 ^fgh (49) Kimchi with 1% Sodium Alginate 0.615 ± 0.021 ^ab (23) 0.409 ± 0.075 ^bcd (64) Kimchi with 1% added powder 0.452 ± 0.055 ^cde (43) 0.320 ± 0.024 ^ef (60) Organic Cabbage Kimchi 0.406 ± 0.053 ^de (49) 0.330 ± 0.017 ^bc (59) ^{One) a ~ h} represent significant differences (p <0.05) in Duncan's multiple range test.

Effect of Juvenile Kimchi from 13 Cabbage Kimchi at 15 ° C until pH 4.3, Survival Effect of HT-29 Colon Cancer Cells by Human MTT Test Treated substance (μl / assay) OD ₅₄₀ 5 10 Control 0.425 ± 0.017 ^a 0.626 ± 0.032 ^a Standardized Chinese Cabbage Kimchi 0.197 ± 0.017 ^bcdef (54) ¹⁾ 0.256 ± 0.040 ^defgh (59) Kimchi with 10.5% red pepper powder 0.168 ± 0.002 ^def (61) 0.236 ± 0.015 ^efgh (62) Kimchi with 5.2% garlic 0.139 ± 0.014 ^f (67) 0.281 ± 0.003 ^cde (55) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.136 ± 0.006 ^f (68) 0.352 ± 0.021 ^b (44) 5% freshly added kimchi 0.152 ± 0.019 ^ef (64) 0.204 ± 0.018 ^h (67) 5% Hearing Added Kimchi 0.182 ± 0.018 ^cdef (57) 0.315 ± 0.019 ^bc (50) Kimchi with 1% Chopi 0.170 ± 0.010 ^def (60) 0.241 ± 0.031 ^efgh (62) Kimchi with 1% Cinnamon Powder 0.243 ± 0.013 ^b (43) 0.270 ± 0.021 ^cdef (57) Kimchi with 1% Ginseng Powder 0.235 ± 0.008 ^bc (45) 0.219 ± 0.004 ^fgh (65) Kimchi with 1% chitosan oligosaccharide 0.216 ± 0.070 ^bcd (49) 0.120 ± 0.046 ⁱ (81) Kimchi with 1% Sodium Alginate 0.223 ± 0.013 ^bcd (48) 0.299 ± 0.004 ^cd (52) Kimchi with 1% added powder 0.210 ± 0.070 ^bcde (51) 0.211 ± 0.260 ^gh (66) Organic Cabbage Kimchi 0.217 ± 0.053 ^f (49) 0.260 ± 0.005 ^defg (59) ^{One) a ~ h} represent significant differences (p <0.05) in Duncan's multiple range test.

Experimental Example 4: anticancer effect of cabbage kimchi juice fermented at 25 ℃

The anticancer effect was investigated in the same manner as in Experimental Example 1, but the cabbage kimchi was fermented at 25 ° C. until pH 4.3, and the juice was used as a sample. Experimental results, as shown in Table 8 when the fermentation at 25 ℃ compared to the cabbage kimchi fermented at 15 ℃ of Experimental Example 2 and the cabbage kimchi juice fermented at 15 ℃ of Experimental Example 2 after 15 days of fermentation at 15 ℃. Showed slightly lower gastric cancer cell survival, and compared with standardized Chinese cabbage kimchi, Chinese cabbage kimchi added with red pepper powder and garlic twice, Chinese cabbage kimchi added with fresh 5%, and organic cabbage kimchi juice showed higher inhibitory effect on gastric cancer cell survival. p <0.05). Standardized Chinese cabbage kimchi showed 29% inhibitory effect at 10µl / assay juice concentration, while kimchi containing 7% red pepper powder + 2.8% garlic, fresh kimchi added 5%, organic cabbage kimchi 62%, 58%, The cancer cell survival inhibitory effect was 49% (p <0.05). In addition, the effect of 13 kinds of cabbage kimchi juice fermented at 25 ° C on the survival of HT-29 human colon cancer cells is different from AGS human gastric cancer cells. Did not look. Compared to the standardized Chinese cabbage kimchi juice, kimchi with 10.5% red pepper powder, kimchi with 7% red pepper powder and garlic, 2.8% kimchi with 5% hearing and kimchi with 1% chokcho and organic cabbage kimchi with 1% choppy and Survival inhibitory effect of HT-29 colon cancer cells (p <0.05). In particular, kimchi added with high red pepper powder showed high inhibitory effect, while standardized Chinese cabbage juice showed 43% inhibitory effect, while cabbage kimchi juice added with red pepper powder and garlic twice each treated with 10µl / assay. The concentration showed much higher inhibitory effect of 72%.

Above, cabbage kimchi dipped in different amounts and types of subsidiary materials from the results of Experimental Examples 1 to 4 were slightly different depending on the fermentation temperature. Chinese cabbage kimchi showed the highest inhibitory effect. Also, Kimchi, Chochu added kimchi and organic cabbage kimchi with high red pepper powder and garlic significantly inhibited the survival of AGS human gastric cancer cells and HT-29 colon cancer cells. Could know.

13 kinds of Chinese cabbage kimchi are fermented at 25 ℃ until pH 4.3, and the juice solution inhibits the survival of human gastric cancer cells Treated substance (μl / assay) OD ₅₄₀ 5 10 Control 0.672 ± 0.07 ^a 0.672 ± 0.07 ^a Standardized Chinese Cabbage Kimchi 0.349 ± 0.034 ^ef (48) ¹⁾ 0.542 ± 0.086 ^b (29) Kimchi with 10.5% red pepper powder 0.376 ± 0.018 ^de (44) 0.226 ± 0.055 ^f (70) Kimchi with 5.2% garlic 0.396 ± 0.075 ^de (41) 0.477 ± 0.045 ^bc (37) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.437 ± 0.030 ^cd (35) 0.287 ± 0.001 ^ef (62) 5% freshly added kimchi 0.355 ± 0.048 ^ef (47) 0.320 ± 0.075 ^def (58) 5% Hearing Added Kimchi 0.349 ± 0.043 ^ef (48) 0.430 ± 0.069 ^bcd (43) Kimchi with 1% Chopi 0.464 ± 0.007 ^c (31) 0.449 ± 0.001 ^bcd (41) Kimchi with 1% Cinnamon Powder 0.396 ± 0.460 ^de (41) 0.516 ± 0.053 ^b (32) Kimchi with 1% Ginseng Powder 0.304 ± 0.011 ^f (55) 0.482 ± 0.126 ^bc (36) Kimchi with 1% chitosan oligosaccharide 0.562 ± 0.026 ^b (16) 0.381 ± 0.055 ^cde (50) Kimchi with 1% Sodium Alginate 0.595 ± 0.017 ^b (12) 0.387 ± 0.116 ^cde (49) Kimchi with 1% added powder 0.580 ± 0.025 ^b (8) 0.380 ± 0.056 ^cde (8) Organic Cabbage Kimchi 0.343 ± 0.026 ^ef (49) 0.390 ± 0.071 ^cde (49) ^{One) a ~ f} represent the significant difference (p <0.05) of Duncan's multiple range test.

The effect of inhibiting the survival of HT-29 human colon cancer cells by MTT assay of the juice obtained by fermenting 13 kinds of Chinese cabbage kimchi to pH 4.3 at 25 ℃ Treated substance (μl / assay) OD540 5 10 Control 0.513 ± 0.02 ^a 0.702 ± 0.007 ^a Standardized Chinese Cabbage Kimchi 0.262 ± 0.031 ^cd (49) ¹⁾ 0.399 ± 0.010 ^b (43) Kimchi with 10.5% red pepper powder 0.231 ± 0.004 ^d (55) 0.193 ± 0.056 ^f (73) Kimchi with 5.2% garlic 0.221 ± 0.050 ^d (57) 0.399 ± 0.069 ^b (43) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.198 ± 0.018 ^de (61) 0.194 ± 0.073 ^f (72) 5% freshly added kimchi 0.196 ± 0.012 ^de (62) 0.291 ± 0.024 ^cde (59) 5% Hearing Added Kimchi 0.234 ± 0.037 ^d (54) 0.245 ± 0.034 ^def (65) Kimchi with 1% Chopi 0.229 ± 0.012 ^d (55) 0.260 ± 0.029 ^cdef (63) Kimchi with 1% Cinnamon Powder 0.358 ± 0.057 ^b (30) 0.317 ± 0.017 ^c (55) Kimchi with 1% Ginseng Powder 0.307 ± 0.037 ^bc (40) 0.316 ± 0.026 ^cd (55) Kimchi with 1% chitosan oligosaccharide 0.336 ± 0.008 ^b (35) 0.259 ± 0.008 ^cdef (63) Kimchi with 1% Sodium Alginate 0.352 ± 0.011 ^b (31) 0.263 ± 0.029 ^cdef (63) Kimchi with 1% added powder 0.310 ± 0.032 ^e (8) 0.255 ± 0.009 ^ef (8) Organic Cabbage Kimchi 0.141 ± 0.028 ^bc (73) 0.237 ± 0.039 ^cdef (66) One) a to f represent significant differences (p <0.05) in Duncan's multiple range test.

Example 4 Antimutagenic Effect of Cabbage Kimchi Methanol Extract

Experimental Example 1: antimutagenic effect by Ames test

The strain used in this experiment was Samonella typhimurium TA100 used by Dr. BN Ames of the University of California, USA. Aflatoxin B _{1, an} indirect mutagen, was purchased from Sigma, USA. In order to activate the indirect mutagen, a mixture of S9, a microsomal enzyme compound of liver, was prepared according to the method of Maron and Ames. For the enzymatic activity of about 200 g of male Sprague-Dawley rats, Aroclor 1254, a mixture of polychlorinated biphenyl (PCB), was intraperitoneally injected at a concentration of 200 mg per 1 mL of corn oil. Liver was removed after 5 days (500 mg / kg). Liver harvested at 45 ° C. aseptic state was washed several times with 0.15 KCl and homogenized in a homogenizer (Potter-Elvehien apparatus, USA) by adding 0.15M KCl solution three times the liver weight. This was centrifuged at 9000 × g for 10 minutes to obtain the S9 fraction, the supernatant, which was dispensed in 1 to 2 mL into a cryogenic tube and rapidly frozen on dry ice, followed by a deep freezer. ). This S9 fraction (10%) was added with MgCl-KCl salt (2%), 1M glucose-6-phosphate (0.5%), 1M NADP (4%), 0.2M phosphate buffer. (pH 7.4) and sterile water were mixed to prepare a S9 mixture. Before the fermented cabbage kimchi methanol extract sample was used in the experiment, a toxicity test was conducted to determine the concentration of the sample within the range of no toxicity. First, dispense 2 mL of the top agar into a sterile cap test tube, add 100 μl of the strain and 100 μl of the diluted sample, lightly vortex, and then minimal agar plate. ) Was aliquoted and solidified, and then cultured at 37 ° C for 24 hours, and then the presence of mutation was determined. Mutation inhibitory effect was calculated by the following formula.

% Inhibition = 100 × [(a-b) / (a-c)]

a is the number of back mutations induced by the mutagen

b is the number of return mutations when the sample is processed

c is the natural reversion mutation in the absence of mutagen and sample.

As a result, as shown in Table 10, 13 kinds of Chinese cabbage kimchi (15 ℃ → 5 ℃ fermentation, pH 4.3) dipped by changing the amount of subsidiary materials or by adding cancer prevention functional material, methanol extract (water-soluble and fat-soluble) The antimutagenic effect on aflatoxin (AFB ₁ ) in Salmonella typhimurium TA100 strain of the material extracted) inhibited the mutagenesis of AFB ₁ by 66% for standardized cabbage kimchi. In comparison, kimchi added with high concentration of garlic (5.2%) and kimchi added with garlic and red pepper powder of 2.8% and 7% showed higher antimutagenic effects of more than 75% (p <0.05). In addition, kimchi added with 5% of fresh and ginseng powder added with 1% of ginseng significantly inhibited mutagenesis. Especially, kimchi added with freshness showed inhibitory effect of 86% (p <0.05). Chinese cabbage kimchi soaked with organic cabbage also showed higher antimutagenic effect than standardized kimchi, which suppressed 80% mutagenesis of AFB ₁ . Therefore, it was found that the antimutagenic effect of kimchi added with garlic at high concentration, kimchi added with fresh and ginseng powder, and organic cabbage kimchi improved compared to standardized cabbage kimchi.

A sub material is added Kimchi ^- methanol extract (2.5mg / plate) antimutagenic effect on AFB ₁ (2.0㎍ / plate) in TA100 strain of Salmonella typhimurium Treatment Return mutation / plate % Inhibition Natural Return Mutation (Spontaneous) AFB ₁ (Control) 118 ± 141099 ± 77 ^a Standardized Chinese Cabbage Kimchi 456 ± 29 ^de 66 Kimchi with 10.5% red pepper powder 449 ± 38 ^de 66 Kimchi with 5.2% garlic 360 ± 50 ^fg 76 7% Red Pepper Powder + 2.8% Garlic Kimchi 357 ± 49 ^fg 76 5% freshly added kimchi 261 ± 5 ^h 86 5% Hearing Added Kimchi 438 ± 16 ^de 68 Kimchi with 1% Chopi 436 ± 17 ^ef 68 Kimchi with 1% Cinnamon Powder 496 ± 73 ^d 62 Kimchi with 1% Ginseng Powder 401 ± 16 ^ef 71 Kimchi with 1% chitosan oligosaccharide 601 ± 16 ^c 51 Kimchi with 1% Sodium Alginate 672 ± 60 ^bc 41 Kimchi with 1% added powder 677 ± 28 ^b 43 Organic Cabbage Kimchi 314 ± 47 ^gh 80 ^* Kimchi samples ^{a ~ h} fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 show a significant difference (p <0.05) in Duncan's multiple range test.

Experimental Example 2: Antimutagenic Effect by SOS Chromotest

In this experimental example, about 90% of the mutants identified in the Ames test of Experimental Example 1 were SOS-inducing agents, and it was reported that there is a quantitative correlation between these two methods. MNNG using SOS chromotest was used. The effects of 13 types of Chinese cabbage kimchi on the inhibition of SOS reactions were investigated for different types and amounts of (N-methyl-N'-nitro-N-nitrosoguanidine).

The strain used in the experiment was E. coli PQ37 (Kangwon University Hamseung City professor) derived from E. coli GC4436, the strain was mixed at 90% glycerol and incubated overnight in L medium and stored at -20 ℃. The mutagenesis was a direct mutagenesis, MNNG was purchased from Aldrich, USA and dissolved in distilled water and used in the experiment.

50 μl of the frozen PQ37 bacteria solution was inoculated in 5 mL L culture medium, shaken overnight at 37 ° C., and then inoculated again in 5 mL L culture medium and shaken for 2 hours until the A ₆₆₀ measurement reached 0.3-0.4. The obtained microbial solution was diluted 1/10 in L culture medium, and 100 µl of each well of a 96-well plate in which 20 µl of a sample prepared by each concentration and a mixture of mutagen were preliminarily dispensed was aliquoted and shaken at 37 ° C for 90 minutes. After inducing the SOS response, on one side, O-nitrophenyl-β-D-galactopyranoside (O-nitrophenyl-β- for measuring activity of β-galactosidase (β-G) 100 μl of D-galactopyraniside (ONPG), and 100 μl of P-nitrophenyl phosphate disodium (PNPP) were added to the other side to measure the activity of alkaline phosphatase (AP). . The color development time was 30 minutes, β-G was 100M of 1.5M Na ₂ CO ₃ , AP was 50M of 1M HCl, and the color development reaction was stopped by enzyme. After 5 minutes, 50μL of 2M Tris buffer was added to AP to HCl. Was neutralized and the absorbance was measured at 420 nm with a spectrophotometer. The measured OD 420 nm measured enzyme value (EU) by Miller's formula.

EU = (1000 × A ₄₂₀ ) / t (min)

As a result, as shown in Table 11, chromotest using MNNG showed a similar effect to that of Ames experimental system. Chinese cabbage kimchi with 10.5% red pepper powder and garlic cabbage kimchi with 5.2% garlic showed higher inhibitory effect on SOS than standardized Chinese cabbage kimchi. Also, standardized Chinese cabbage with 1% chopi kimchi, sodium alginate and kimchi powder added It showed higher inhibitory effect than kimchi (p <0.05). In particular, organic cabbage kimchi showed the highest inhibitory effect on SOS reaction, and the inhibitory effect of 55% was observed compared to the antimutagenic effect of 33% of standardized cabbage kimchi. Therefore, red pepper powder, garlic, chopi added kimchi and organic cabbage kimchi have high antimutagenic effect, which can increase antimutagenic effect by adding red pepper powder and garlic to organic cabbage.

Antimutagenic effect on N-methyl-N'- nitro-N -nitrosoguanidine (MNNG, 40ng / assay) in the E.coli strain PQ37 of the sub material is added Kimchi ^- methanol extract Treatment β-galactosidase Unit (β) Alkaline phosphatase Unit (β) / (ρ) SOS Inducer Inhibition Ratio (%) Spontaneous Control Group (MNNG) 22.388.9 ^a 14.613.9 1.526.41 1.004.22 Standardized Chinese Cabbage Kimchi 73.9 ^f 15.4 4.79 3.15 33 Kimchi with 10.5% red pepper powder 65.8 ^h 15.5 4.25 2.80 44 Kimchi with 5.2% garlic 65.5 ^h 15.6 4.19 2.76 45 7% Red Pepper Powder + 2.8% Garlic Kimchi 63.3 ⁱ 15.0 4.22 2.78 45 5% freshly added kimchi 72.7 ^g 14.9 4.87 3.20 32 5% Hearing Added Kimchi 81.5 ^c 16.3 5.01 3.30 29 Kimchi with 1% Chopi 64.7 ^h 15.1 4.28 2.82 44 Kimchi with 1% Cinnamon Powder 79.1 ^d 16.0 4.95 3.26 30 Kimchi with 1% Ginseng Powder 87.3 ^b 16.9 5.17 3.40 26 Kimchi with 1% chitosan oligosaccharide 76.8 ^e 16.1 4.76 3.13 34 Kimchi with 1% Sodium Alginate 71.6 ^g 16.2 4.42 2.91 41 Kimchi with 1% added powder 70.5 ^g 15.9 4.44 2.92 40 Organic Cabbage Kimchi 59.5 ^j 15.9 3.74 2.46 55 ^* Kimchi samples ^{a ~ j} fermented at 15 ℃ for 1 day and then fermented to pH 4.3 at 5 ℃ show significant difference (p <0.05) in Duncan's multiple range test.

Example 5: Anticancer Effect of Cabbage Kimchi Methanol Extract

Experimental Example 1: Anticancer Effect of Cabbage Kimchi Methanol Extract in SRB Method

Cancer cells were seeded at 40,000 cells / mL per well in a 96 well plate, and after incubation for 24 hours, when cells were attached to the plate, 100 μl of sample extract was added and then cultured in a 37 ° C. 5% CO ₂ incubator. At this time, only a sample containing 10% FCS was added to a blank and a DMSO was added to the control instead of cells and the sample. After 48 hours of incubation, the medium was removed, washed once with PBS, and then refrigerated at 4 ° C by adding 50% TCA. After 1 hour, TCA was removed, washed five times with distilled water, dried at room temperature, and then stained for 30 minutes by adding 100 µl of 0.4% sulforhodamine B. Next, washed 5 times with 1% acetic acid and dried again at room temperature, and 150 μl of 0.01M Tris base was added, and the absorbance was measured at 510 nm. As a result, as shown in Table 12, kimchi added 7% and 2.8% of red pepper powder and garlic at 0.05 mg / mL treatment concentration showed more than 50% of cancer cell survival inhibition. On the other hand, at the concentration of 0.1mg / mL, red pepper powder 10.5%, garlic 5.2%, red pepper powder and garlic with 7% and 2.8% kimchi, 1% chopi added kimchi, 1% added powder kimchi and organic cabbage kimchi 50% Abnormal AGS inhibited the survival of human gastric cancer cells (p <0.05). As shown in Table 13, the results of SRB assay on 13 kinds of kimchi methanol extracts using HT-29 human colon cancer cells showed that kimchi and red pepper powder added three times (10.5%) of red pepper powder compared to standard kimchi. Chinese cabbage kimchi containing 7% and 2.8% garlic and garlic extracts inhibited the survival of high HT-29 colon cancer cells (p <0.05). In addition, it was found that kimchi and organic cabbage kimchi added with Gad, Chopi, Sodium Alginate, and powdered cabbage inhibited the survival of colon cancer cells by more than 50% (p <0.05). Table 14 shows the results of 13 kinds of Chinese cabbage kimchi (15 ℃ → 5 ℃ fermentation, pH 4.3) methanol extract of HL-60 human blood cancer cell survival inhibition effect by SRB assay. Kimchi with 10.5% red pepper powder, Kimchi with 7% red pepper powder and garlic, and Kimchi with 1% chopi and 1% Choppi and 0.1% organic cabbage kimchi at 0.1 mg / mL, respectively. The survival of blood cancer cells was further suppressed (p <0.05). This is similar to the results of other human cancer cells, it was found that the red pepper powder, garlic, chopi and organic cabbage is a subsidiary material that can improve the anticancer function in the Chinese cabbage kimchi. Table 15 shows the survival inhibition effect of 13 kinds of Chinese cabbage kimchi methanol extract on MG-63 human bone cancer cells compared to other human cancer cells, namely stomach cancer, colon cancer, and blood cancer. Only cabbage kimchi added with red pepper powder 7% and garlic 2.8% showed an inhibitory effect of 40% at the treatment concentration of 0.1 mg / mL (p <0.05).

Effect of Substances Added Cabbage Kimchi ^* AGS Human Gastric Cancer Cell Survival Inhibitory Effect by Methanol Extract Treated product (mg / ml) OD ₅₁₀ 0.05 0.1 Control 0.757 ± 0.036 ^a 0.757 ± 0.036 Standardized Chinese Cabbage Kimchi 0.691 ± 0.076 ^ab (9) ¹⁾ 0.472 ± 0.006 ^b (38) Kimchi with 10.5% red pepper powder 0.667 ± 0.067 ^ab (12) 0.328 ± 0.068 ^cde (57) Kimchi with 5.2% garlic 0.616 ± 0.105 ^ab (19) 0.308 ± 0.017 ^de (59) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.339 ± 0.047 ^c (55) 0.251 ± 0.013 ^e (67) 5% freshly added kimchi 0.411 ± 0.025 ^c (46) 0.399 ± 0.093 ^bcd (47) 5% Hearing Added Kimchi 0.653 ± 0.021 ^ab (14) 0.389 ± 0.076 ^bcd (49) Kimchi with 1% Chopi 0.720 ± 0.050 ^ab (5) 0.200 ± 0.005 ^e (74) Kimchi with 1% Cinnamon Powder 0.691 ± 0.049 ^ab (9) 0.434 ± 0.012 ^bc (43) Kimchi with 1% Ginseng Powder 0.687 ± 0.114 ^ab (9) 0.516 ± 0.161 ^b (32) Kimchi with 1% chitosan oligosaccharide 0.389 ± 0.109 ^c (49) 0.519 ± 0.075 ^b (31) Kimchi with 1% Sodium Alginate 0.393 ± 0.041 ^c (48) 0.397 ± 0.066 ^bcd (48) Kimchi with 1% added powder 0.583 ± 0.008 ^b (23) 0.201 ± 0.027 ^e (73) Organic Cabbage Kimchi 0.663 ± 0.107 ^ab (12) 0.294 ± 0.003 ^de (61) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ e} represent significant differences (p <0.05) in Duncan's multiple range test.

Effect of Substances Added Cabbage Kimchi ^* Survival Inhibitory Effect of HT-29 Human Colon Cancer Cell by Methanol Extract SRB Method Treated product (mg / ml) OD ₅₁₀ 0.05 0.1 Control 0.718 ± 0.072 ^a 0.638 ± 0.072 ^a Standardized Chinese Cabbage Kimchi 0.443 ± 0.030 ^c (38) ²⁾ 0.460 ± 0.038 ^bc (36) Kimchi with 10.5% red pepper powder 0.479 ± 0.005 ^c (33) 0.196 ± 0.047 ^f (73) Kimchi with 5.2% garlic 0.711 ± 0.021 ^a (1) 0.498 ± 0.029 ^b (31) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.402 ± 0.046 ^c (44) 0.038 ± 0.035 ^g (95) 5% freshly added kimchi 0.590 ± 0.008 ^b (18) 0.331 ± 0.032 ^e (54) 5% Hearing Added Kimchi 0.439 ± 0.024 ^c (39) 0.514 ± 0.031 ^b (28) Kimchi with 1% Chopi 0.407 ± 0.067 ^c (43) 0.329 ± 0.047 ^e (54) Kimchi with 1% Cinnamon Powder 0.596 ± 0.005 ^b (17) 0.412 ± 0.022 ^cd (35) Kimchi with 1% Ginseng Powder 0.703 ± 0.026 ^a (2) 0.482 ± 0.042 ^b (33) Kimchi with 1% chitosan oligosaccharide 0.717 ± 0.062 ^a (0) 0.360 ± 0.006 ^de (50) Kimchi with 1% Sodium Alginate 0.668 ± 0.027 ^ab (7) 0.308 ± 0.044 ^e (57) Kimchi with 1% added powder 0.394 ± 0.030 ^c (45) 0.342 ± 0.007 ^e (52) Organic Cabbage Kimchi 0.448 ± 0.033 ^c (38) 0.345 ± 0.044 ^e (52) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ g} represent significant differences (p <0.05) in Duncan's multiple range test.

Various Chinese Cabbage Kimchi with Substances ^* Survival of HL-60 Human Blood Cancer Cells by Methanol Extract of SRB Treated product (mg / ml) OD ₅₁₀ 0.05 0.1 Control 0.872 ± 0.012 ^a 0.872 ± 0.012 ^a Standardized Chinese Cabbage Kimchi 0.635 ± 0.023 ^b (27) ¹⁾ 0.198 ± 0.079 ^cdef (77) Kimchi with 10.5% red pepper powder 0.425 ± 0.016 ^def (51) 0.050 ± 0.022 ^g (94) Kimchi with 5.2% garlic 0.520 ± 0.060 ^cd (40) 0.221 ± 0.030 ^cde (75) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.366 ± 0.016 ^f (49) 0.146 ± 0.132 ^defg (83) 5% freshly added kimchi 0.449 ± 0.032 ^def (49) 0.287 ± 0.030 ^cd (67) 5% Hearing Added Kimchi 0.362 ± 0.065 ^f (59) 0.230 ± 0.023 ^cde (74) Kimchi with 1% Chopi 0.335 ± 0.087 ^ef (62) 0.079 ± 0.028 ^fg (91) Kimchi with 1% Cinnamon Powder 0.512 ± 0.037 ^cde (41) 0.271 ± 0.081 ^cde (69) Kimchi with 1% Ginseng Powder 0.495 ± 0.035 ^cde (43) 0.423 ± 0.030 ^b (52) Kimchi with 1% chitosan oligosaccharide 0.410 ± 0.057 ^def (53) 0.328 ± 0.077 ^bc (62) Kimchi with 1% Sodium Alginate 0.595 ± 0.022 ^bc (32) 0.296 ± 0.057 ^cde (65) Kimchi with 1% added powder 0.462 ± 0.052 ^def (47) 0.145 ± 0.119 ^defg (83) Organic Cabbage Kimchi 0.493 ± 0.088 ^cde (44) 0.128 ± 0.042 ^efg (85) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ g} represent significant differences (p <0.05) in Duncan's multiple range test.

Various Chinese Cabbage Kimchi Added Substances ^* Survival of MG-63 Human Bone Cancer Cells by SRB Test of Methanol Extract Treated product (mg / ml) OD ₅₁₀ 0.05 0.1 Control 0.501 ± 0.018 ^a 0.501 ± 0.018 ^a Standardized Chinese Cabbage Kimchi 0.392 ± 0.082 ^bcd (22) ¹⁾ 0.366 ± 0.014 ^ef (27) Kimchi with 10.5% red pepper powder 0.372 ± 0.001 ^bcde (26) 0.441 ± 0.006 ^bcd (12) Kimchi with 5.2% garlic 0.441 ± 0.035 ^ab (12) 0.452 ± 0.034 ^abcd (10) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.375 ± 0.050 ^bcde (25) 0.090 ± 0.005 ^h (40) 5% freshly added kimchi 0.355 ± 0.007 ^cdef (29) 0.299 ± 0.002 ^g (18) 5% Hearing Added Kimchi 0.441 ± 0.044 ^ab (12) 0.411 ± 0.053 ^cde (20) Kimchi with 1% Chopi 0.258 ± 0.027 ^g (49) 0.401 ± 0.039 ^de (5) Kimchi with 1% Cinnamon Powder 0.404 ± 0.042 ^bc (19) 0.474 ± 0.040 ^ab 6) Kimchi with 1% Ginseng Powder 0.351 ± 0.013 ^cdef (30) 0.469 ± 0.055 ^ab (28) Kimchi with 1% chitosan oligosaccharide 0.351 ± 0.103 ^cdef (30) 0.363 ± 0.017 ^ef (9) Kimchi with 1% Sodium Alginate 0.324 ± 0.069 ^defg (35) 0.456 ± 0.008 ^abc (8) Kimchi with 1% added powder 0.311 ± 0.005 ^efg (38) 0.462 ± 0.027 ^abc (20) Organic Cabbage Kimchi 0.295 ± 0.029 ^fg (41) 0.349 ± 0.017 ^f (30) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ h} represent significant differences (p <0.05) in Duncan's multiple range test.

Experimental Example 2: Anticancer Effect of Cabbage Kimchi Methanol Extract by MTT Method

In this experimental example, the anticancer effect was carried out by the same method as the MTT assay performed in Example 3, and the difference from Example 3 was that the Chinese cabbage kimchi methanol extract was used without using the cabbage kimchi juice as a sample. Experimental results, as shown in Table 16, the kimchi, kimchi, kimchi, chitosan oligosaccharide added kimchi, added powdered kimchi and organic cabbage kimchi added to the high concentration of red pepper powder and garlic, similar to the SRB assay result of Experiment 1 The survival of gastric cancer cells was significantly inhibited (p <0.05). In particular, kimchi added with red pepper powder and garlic and 2.8% kimchi, kimchi added with chopi and organic cabbage kimchi showed a high inhibitory effect on cancer cell survival by more than 90%. This result was consistent with the fact that in vitro anticancer experiment using the juice of Example 3 kimchi, chopi added kimchi and organic cabbage kimchi with high concentration of red pepper powder and garlic significantly inhibited the survival of gastric cancer cells. Therefore, it was found that cancer prevention kimchi can be developed in consideration of functionality, functionality, and shelf life. Table 17 shows the results of comparing the effect of HT-29 on human colon cancer cell survival by MTT assay. Kimchi (10.5%), red pepper powder and garlic added twice as much as red pepper powder compared to standardized Chinese cabbage kimchi. One kimchi and organic cabbage kimchi showed high inhibitory effect on colon cancer cell survival. In addition, fresh kimchi, sodium alginate, and added kimchi showed high inhibitory effect (p <0.05). Table 18 shows the survival inhibition effect of HL-60 human blood cancer cells of 13 kinds of Chinese cabbage kimchi samples through MTT assay. Kimchi, chochu added kimchi and organically added pepper powder and garlic at high concentrations as in SRB assay of Experimental Example 1 Chinese cabbage kimchi inhibited the survival of blood cancer cells more than standard cabbage kimchi (p <0.05). In particular, cabbage kimchi added with red pepper powder and garlic twice as standardized cabbage kimchi showed more than 50% inhibitory effect even at the concentration of 0.05mg / ml, and very high inhibition of hematologic cancer cell survival of 91% at 0.1mg / ml. The effect was shown. Table 19 also shows the results of comparing the effects of 13 kinds of Chinese cabbage kimchi samples on the survival of MG-63 bone cancer cancer cells by MTT assay. At 0.1 mg / ml treatment, only kimchi added red pepper powder and garlic twice as much as standardized cabbage kimchi and kimchi added 1% of chopi showed the inhibitory effect on survival of 50% bone cancer cancer cells (p <0.05). ).

Inhibitory Effect of AGS Human Gastric Cancer Cell Survival by MTT Assay on Chinese Cabbage Kimchi Extract Treated product (mg / ml) OD ₅₄₀ 0.05 0.1 Control 0.747 ± 0.004 ^a 0.747 ± 0.004 ^a Standardized Chinese Cabbage Kimchi 0.491 ± 0.087 ^bc (34) ¹⁾ 0.328 ± 0.045 ^bc (57) Kimchi with 10.5% red pepper powder 0.393 ± 0.112 ^c (47) 0.152 ± 0.055 ^d (80) Kimchi with 5.2% garlic 0.511 ± 0.122 ^bc (32) 0.071 ± 0.017 ^de (91) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.418 ± 0.023 ^c (44) 0.074 ± 0.029 ^de (90) 5% freshly added kimchi 0.517 ± 0.041 ^b (31) 0.395 ± 0.069 ^bc (47) 5% Hearing Added Kimchi 0.513 ± 0.048 ^bc (31) 0.380 ± 0.009 ^bc (49) Kimchi with 1% Chopi 0.400 ± 0.063 ^c (47) 0.015 ± 0.010 ^e (98) Kimchi with 1% Cinnamon Powder 0.432 ± 0.045 ^bc (42) 0.455 ± 0.013 ^b (39) Kimchi with 1% Ginseng Powder 0.453 ± 0.074 ^bc (39) 0.331 ± 0.038 ^c (56) Kimchi with 1% chitosan oligosaccharide 0.444 ± 0.074 ^bc (41) 0.114 ± 0.038 ^de (85) Kimchi with 1% Sodium Alginate 0.400 ± 0.011 ^c (47) 0.462 ± 0.025 ^b (38) Kimchi with 1% added powder 0.132 ± 0.006 ^d (82) 0.082 ± 0.026 ^de (89) Organic Cabbage Kimchi 0.449 ± 0.046 ^bc (40) 0.064 ± 0.021 ^de (91) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ e} represent the significant difference (p <0.05) of Duncan's multiple range test.

Effect of Methanol Extract of Chinese Cabbage Kimchi with Substances ^* Treated product (mg / ml) OD ₅₄₀ 0.05 0.1 Control 0.434 ± 0.060 ^a 0.594 ± 0.001 ^a Standardized Chinese Cabbage Kimchi 0.385 ± 0.019 ^b (11) ¹⁾ 0.257 ± 0.027 ^cd (60) Kimchi with 10.5% red pepper powder 0.304 ± 0.042 ^c (30) 0.124 ± 0.016 ^ef (79) Kimchi with 5.2% garlic 0.397 ± 0.042 ^b (9) 0.251 ± 0.037 ^cd (58) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.297 ± 0.036 ^c (36) 0.081 ± 0.021 ^f (86) 5% freshly added kimchi 0.297 ± 0.036 ^c (32) 0.189 ± 0.016 ^de (68) 5% Hearing Added Kimchi 0.338 ± 0.010 ^c (22) 0.427 ± 0.031 ^b (28) Kimchi with 1% Chopi 0.390 ± 0.029 ^b (10) 0.292 ± 0.067 ^c (51) Kimchi with 1% Cinnamon Powder 0.377 ± 0.012 ^b (13) 0.287 ± 0.053 ^c (52) Kimchi with 1% Ginseng Powder 0.393 ± 0.046 ^b (9) 0.430 ± 0.024 ^b (28) Kimchi with 1% chitosan oligosaccharide 0.142 ± 0.073 ^a (2) 0.292 ± 0.026 ^c (51) Kimchi with 1% Sodium Alginate 0.438 ± 0.061 ^ab (1) 0.134 ± 0.016 ^ef (77) Kimchi with 1% added powder 0.276 ± 0.075 ^c (36) 0.148 ± 0.010 ^ef (75) Organic Cabbage Kimchi 0.250 ± 0.036 ^c (42) 0.089 ± 0.039 ^f (85) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a to f} represent significant differences (p <0.05) in Duncan's multiple range test.

Effect of Substances Added Cabbage Kimchi ^* MHL Assay on MHL Assay for Viability of Human Blood Cancer Cells Treated product (mg / ml) OD ₅₄₀ 0.05 0.1 Control 0.507 ± 0.078 ^a 0.507 ± 0.078 ^a Standardized Chinese Cabbage Kimchi 0.418 ± 0.037 ^abc (18) ¹⁾ 0.109 ± 0.015 ^ef (79) Kimchi with 10.5% red pepper powder 0.302 ± 0.002 ^def (40) 0.089 ± 0.017 ^ef (82) Kimchi with 5.2% garlic 0.395 ± 0.047 ^bcd (22) 0.081 ± 0.027 ^f (84) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.222 ± 0.030 ^f (56) 0.044 ± 0.011 ^f (91) 5% freshly added kimchi 0.459 ± 0.043 ^ab (10) 0.277 ± 0.055 ^c (45) 5% Hearing Added Kimchi 0.430 ± 0.111 ^ab (15) 0.067 ± 0.039 ^f (87) Kimchi with 1% Chopi ^0.361 ± 0.007 ^bcde (28) 0.050 ± 0.059 ^f (90) Kimchi with 1% Cinnamon Powder 0.400 ± 0.023 ^abcd (21) 0.363 ± 0.008 ^b (28) Kimchi with 1% Ginseng Powder 0.440 ± 0.040 ^bcd (13) 0.234 ± 0.039 ^cd (53) Kimchi with 1% chitosan oligosaccharide 0.409 ± 0.012 ^abcd (19) 0.215 ± 0.061 ^cd (58) Kimchi with 1% Sodium Alginate 0.463 ± 0.118 ^ab (9) 0.404 ± 0.029 ^b (20) Kimchi with 1% added powder 0.316 ± 0.093 ^cdef (37) 0.161 ± 0.066 ^de (68) Organic Cabbage Kimchi 0.263 ± 0.009 ^ef (45) 0.099 ± 0.056 ^ef (81) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a to f} represent significant differences (p <0.05) in Duncan's multiple range test.

Effect of Substances Addition on Cabbage Kimchi ^* Methanol Extracts by MTT Assay Treated product (mg / ml) OD ₅₄₀ 0.05 0.1 Control 0.622 ± 0.05 ^a 0.622 ± 0.05 ^a Standardized Chinese Cabbage Kimchi 0.465 ± 0.001 ^bc (25) ¹⁾ 0.450 ± 0.043 ^bcde (28) Kimchi with 10.5% red pepper powder 0.275 ± 0.047 ^d (56) 0.331 ± 0.007 ^cde (47) Kimchi with 5.2% garlic 0.413 ± 0.016 ^bc (34) 0.481 ± 0.066 ^bcd (23) 7% Red Pepper Powder + 2.8% Garlic Kimchi 0.422 ± 0.047 ^bc (32) 0.294 ± 0.020 ^e (53) 5% freshly added kimchi 0.426 ± 0.033 ^bc (32) 0.365 ± 0.080 ^cde (41) 5% Hearing Added Kimchi 0.437 ± 0.055 ^bc (30) 0.521 ± 0.086 ^abc (16) Kimchi with 1% Chopi 0.417 ± 0.130 ^bc (33) 0.306 ± 0.185 ^de (51) Kimchi with 1% Cinnamon Powder 0.590 ± 0.124 ^a (5) 0.598 ± 0.162 ^a (4) Kimchi with 1% Ginseng Powder 0.358 ± 0.130 ^cd (42) 0.436 ± 0.134 ^cde (30) Kimchi with 1% chitosan oligosaccharide 0.469 ± 0.082 ^bc (25) 0.500 ± 0.109 ^abc (20) Kimchi with 1% Sodium Alginate 0.507 ± 0.013 ^ab (19) 0.410 ± 0.040 ^cde (34) Kimchi with 1% added powder 0.532 ± 0.038 ^ab (15) 0.384 ± 0.052 ^cde (38) Organic Cabbage Kimchi 0.448 ± 0.004 ^bc (24) 0.371 ± 0.062 ^cde (40) ^* Kimchi fermented at 15 ℃ for 1 day and then fermented at 5 ℃ to pH 4.3 ^{1) a ~ e} represent significant differences (p <0.05) in Duncan's multiple range test.

Experimental Example 3: Inhibitory Effect of Chinese Cabbage Kimchi Methanol Extract on Cancer Cell Growth

In this experiment, AGS human gastric cancer cells, HT-29 human colon cancer cells, and MG-63 human osteosarcoma cells were cultured in the same manner as in the culture method of Example 2, and then centrifuged to distribute the integrated cancer cells evenly. After mixing and inoculating (seeding) in a 24 well plate at a concentration of 20,000 cells / mL and incubating overnight, confirming that the cancer cells adhered to the plate, the sample was added to the medium containing 10% FCS, and the culture solution was changed every other day at 37 ° C., Incubated in a 5% CO ₂ incubator. The fermented cabbage kimchi methanol extract sample was replaced with fresh medium added by concentration of 10 μl / mL medium every 2 days, and after 6 days of culture, the grown cells were separated by 0.05% trypsin-0.02% EDTA enzyme and hemoglobin counted. It was measured by a cytometer (hemocytometer) to observe the cancer cell growth inhibitory effect compared to the control. As shown in Table 20, as in the SRB and MTT assays of Experimental Example 1 and Experimental Example 2, kimchi added with red pepper powder, garlic and kimchi added with chopi and organic cabbage kimchi inhibited the growth of gastric cancer cells. It was found that the effect was higher than the standardized Chinese cabbage kimchi (p <0.05). In particular, kimchi added with 1% chopi was observed as 8 × 10 ⁴ / mL (83%) compared to the control cell number of 48 × 10 ⁴ / mL, indicating that growth was significantly inhibited. In addition, Table 21 shows the results of examining the growth inhibitory effect of HT-29 colon cancer cells after culturing for 6 days while processing kimchi samples, adding 10.5% red pepper powder, 7% red pepper powder, 2.8% garlic added kimchi and 1% chopi Kimchi and organic cabbage kimchi significantly inhibited the growth of HT-29 colon cancer cells (p <0.05). In particular, kimchi supplemented with red pepper powder and garlic twice inhibited the growth of colon cancer cells most significantly, and the control group reduced 9 × 10 ⁴ / mL (61%) compared to the cell count of 23 × 10 ⁴ / mL. there was.

Through the above SRB, MTT assay and cancer cell growth inhibitory effect, kimchi added with red pepper powder and garlic, kimchi and organic cabbage kimchi added 2-3 times compared to standardized cabbage kimchi, and survival and growth of AGS human gastric cancer cells and HT-29 colon cancer cells It can be seen that suppressing.

The addition of sub material cabbage ^* growth inhibitory effect on human gastric cancer cell AGS of methanol extract Treatment Cell number (× 10 ⁴ / ml) Inhibition Ratio (%) Control 48.3 ^a 0 Standardized Chinese Cabbage Kimchi 18.3 ^cd 62.1 Kimchi with 10.5% red pepper powder 16.0 ^d 66.9 Kimchi with 5.2% garlic 15.3 ^def 68.3 Kimchi with 7% red pepper powder + 2.8% garlic 13.0 ^ef 73.1 5% freshly added kimchi 23.0 ^b 52.4 5% Hearing Added Kimchi 16.3 ^de 66.3 Kimchi with 1% Chopi 8.3 ^g 82.8 Kimchi with 1% Cinnamon Powder 16.3 ^de 66.3 Kimchi with 1% Ginseng Powder 16.0 ^de 66.9 Kimchi with 1% chitosan oligosaccharide 19.0 ^cd 66.7 Kimchi with 1% Sodium Alginate 13.7 ^ef 71.6 Kimchi with 1% added powder 20.7 ^bc 57.1 Organic Cabbage Kimchi 12.0 ^f 75.2 ^* Kimchi sample ^{a ~ g} Duncan's multiple range test fermented at 15 ℃ for 1 day and then at pH 4.3 at 5 ℃ showed significant difference (p <0.05).

Growth inhibitory effect on HT-29 human colon carcinoma cells of the addition of sub material Kimchi ^- methanol extract Treatment Cell number (× 10 ⁴ / ml) Inhibition Ratio (%) Control 23 ^a 0 Standardized Chinese Cabbage Kimchi 16.0 ^b 30.4 Kimchi with 10.5% red pepper powder 9.5 ^efg 58.7 Kimchi with 5.2% garlic 12.5 ^cde 45.7 Kimchi with 7% red pepper powder + 2.8% garlic 7.5 ^g 67.4 5% freshly added kimchi 14.5 ^bcd 37.0 5% Hearing Added Kimchi 12.0 ^def 47.8 Kimchi with 1% Chopi 9.0 ^fg 60.9 Kimchi with 1% Cinnamon Powder 14.0 ^bcd 39.1 Kimchi with 1% Ginseng Powder 14.0 ^bcd 39.1 Kimchi with 1% chitosan oligosaccharide 15.5 ^bc 32.6 Kimchi with 1% Sodium Alginate 11.5 ^def 50.0 Kimchi with 1% added powder 10.5 ^efg 54.3 Organic Cabbage Kimchi 10.5 ^efg 54.3 ^* Kimchi sample ^{a ~ g} Duncan's multiple range test fermented at 15 ℃ for 1 day and then at pH 4.3 at 5 ℃ showed significant difference (p <0.05).

From the results of Experimental Examples 1, 2 and 3, 13 kinds of Chinese cabbage kimchi extracts with different types and amounts of subsidiary materials significantly inhibited the survival of AGS human gastric cancer cells, HT-29 colon cancer cells and HL-60 human blood cancer cells. On the other hand, MG-63 human bone cancer cells showed a somewhat lower inhibitory effect than other human cancer cells. Kimchi added with red pepper powder and garlic, kimchi added with chopi and organic cabbage kimchi had much higher inhibitory effect than standardized cabbage kimchi. Although ginseng powder, sodium alginate and chitosan oligosaccharides, known as functional foods, are known for their anticancer functionality, when added to kimchi, they did not show much effect in enhancing antimutation and anticancer functionality. Kimchi added with red pepper powder, garlic and chopi or using organic cabbage was found to be more functional.

In addition, kimchi fermented at low temperature had high anti-cancer effects. In experiments using AGS human gastric cancer cells, kimchi fermented at 15 ° C. for 1 day was more effective. This is the same trend as in the previous experiment. For HT-29 human colon cancer cells, kimchi fermented at 15 ° C and 25 ° C showed high cancer cell survival and suppression effect. However, since kimchi fermented at low temperature had a good taste and had a high effect on gastric cancer, it was fermented at 15 ° C. for 1 day and then fermented at 5 ° C. Antimutagenic and anticancer effects depend on the extraction method, concentration and fermentation temperature of the sample, and the experimental method, but kimchi fermented from low temperature (5 ℃) to ripening season (pH 4.3) is the highest.

Example 6 Recipe Adjustment of Chinese Cabbage Kimchi for Anticancer Function Enhancement

In this embodiment, according to the results obtained in the above 1 to 5 to prepare the Chinese cabbage kimchi with the recipe of controlling the concentration of red pepper powder, garlic, chopi, gat with the best anticancer activity, in vitro antimutagenic and anticancer effect and in vivo The effect of nitric oxide production on mouse macrophages in rats was investigated. That is, the same standardized cabbage kimchi (K-1) and organic cabbage kimchi (K-2) added with red pepper powder and garlic 7%, 2.8%, respectively, 7%, 2.8% , Organic cabbage kimchi (K-3) added with 0.1%, red pepper powder, garlic, chopi, and fresh cabbage kimchi (K-4) added with 7%, 2.8%, 0.1% and 5%, respectively, It was fermented at 15 ° C. for 1 day and fermented at 5 ° C. until the pH was 4.3, thereby extracting the mixture with juice and methanol. This juice and methanol extract were prepared as a sample to confirm the antimutagenic and anticancer effect in vitro and the effect on nitric oxide production of mouse macrophages in vivo.

Example 7 Antimutagenic Effect of Cabbage Kimchi Methanol Extract

Experimental Example 1 antimutagenic effect by Ames test

Table 22 shows standardized Chinese cabbage kimchi (K-1), Chinese cabbage kimchi added with 7% red pepper powder and 2.8% garlic (K-2), 7% red pepper powder, 2.8% garlic and 0.1% chopi powder (K-3) ), 7% red pepper powder, 2.8% garlic, 0.1% chopi powder, and 5% freshly added cabbage kimchi (K-4) were prepared to investigate the antimutagenic effect in the Ames test. At 1.25mg / plate addition concentration, 7% red pepper powder, 2.8% garlic, 0.1% chopi powder, and 5% freshly added cabbage kimchi (K-4) showed 65% higher incidence and standardization at 2.5 mg / plate concentration. Chinese cabbage kimchi (K-1) showed 45% antimutagenic effect, but all of the enhanced kimchi showed higher (70% or more) antimutagenic effect against AFB ₁ , and the addition of pepper, garlic, chopped powder, and fresh It has been shown that the use of Chinese cabbage can enhance the antimutagenic effect of Chinese cabbage kimchi.

Antimutagenic effect for the addition of sub material cabbage ^* in Salmonella typhimurium TA100 strain in the methanol extract to AFB ₁ (2.0㎍ / plate) Kimchi Sample Mutagenesis / plate (mg / plate) 1.25 2.5 Natural Return Mutation 105 ± 6 AFB _! (Control) 706 ± 3 ^a K-1 422 ± 3 ^cd (47) ¹ 436 ± 45 ^b (45) K-2 670 ± 33 ^b (19) 286 ± 36 ^c (70) K-3 458 ± 144 ^c (41) 232 ± 53 ^c (79) K-4 317 ± 83 ^d (65) 271 ± 4 ^c (73) ^* Kimchi sample fermented at 15 ℃ for 1 day and then fermented to pH 4.3 at 5 ℃ ¹⁾ Inhibition rate K-1: Standardized cabbage kimchi K-2: Organic cabbage kimchi with 7% red pepper powder + 2.8% garlic K-3 : 7% red pepper powder + 2.8% garlic + 0.1% organic cabbage kimchi added K-4: 7% red pepper powder + 2.8% garlic + 0.1% chopi + 5% fresh organic cabbage kimchi

Example 8 anticancer effect of Chinese cabbage kimchi juice

Experimental Example 1: anticancer enhancement effect by MTT assay

Table 23 shows standardized Chinese cabbage kimchi (K-1), Chinese cabbage kimchi added with 7% red pepper powder and 2.8% garlic (K-2), 7% red pepper powder, 2.8% garlic and 0.1% chopped red cabbage kimchi (K-3) In vitro anticancer activity of 4 kinds of Chinese cabbage kimchi juice (K-4) added with 7% red pepper powder, 2.8% garlic, 0.1% chopi powder and 5% fresh cabbage kimchi juice against AGS human gastric cancer cells . At the treatment concentration of 1.0 μl / assay, 7% red pepper powder, 2.8% garlic, 0.1% chopi powder, and 5% freshly added cabbage kimchi (K-4) showed higher in vitro anticancer activity, but at 2.5 μl / assay While standardized kimchi showed 36% of AGS gastric cancer cell inhibition effect, all three types of anti-cancer active kimchi showed much higher inhibitory effect of more than 60%, and it was best to add red pepper powder, garlic, chopi powder, and fresh cabbage and use organic cabbage. It has been shown that Chinese cabbage kimchi may enhance the in vitro anticancer effect.

Effect of Substances Added Cabbage Kimchi ^* MGS Assay on the Viability of AGS Human Gastric Cancer Cells Treatment (µl / assay) OD ₅₄₀ 1.0 2.5 control 0.738 ± 0.016 ^a 0.714 ± 0.017 ^a K-1 0.525 ± 0.016 ^cd (45) ¹⁾ 0.574 ± 0.016 ^b (36) K-2 0.615 ± 0.014 ^b (26) 0.445 ± 0.014 ^c (67) K-3 0.537 ± 0.018 ^c (44) 0.496 ± 0.035 ^c (61) K-4 0.498 ± 0.014 ^d (55) 0.449 ± 0.005 ^c (63) ^* Kimchi sample fermented at 15 ° C. for 1 day and then fermented to pH 4.3 at 5 ° C. ¹⁾ Inhibition rate K-1: Standardized Chinese cabbage kimchi K-2: Organic cabbage kimchi with 7% red pepper powder + 2.8% garlic K-3 : 7% red pepper powder + 2.8% garlic + 0.1% organic cabbage kimchi added K-4: 7% red pepper powder + 2.8% garlic + 0.1% chopi + 5% fresh organic cabbage kimchi

Example 9 Effect of Anticancer Activity Enhancing Kimchi on Nitric Oxide Production in Mouse Macrophages

Nitric oxide (NO) is a reactive nitrogen intermediate (RNI) produced by macrophages, neutrophils, vascular endothelial cells, synovial hyperplasia, synovial fibroblasts, and chondrocytes under the influence of cytokines or microorganisms. Nitric oxide synthase (NOS) combines with oxygen to oxidize L-arginine to produce NO. The produced NO is known to act as a vasodilator, neurotransmitter, immune modulator, and antimicrobial or anticancer activity in vivo. Standardized Chinese cabbage kimchi (K-1), Chinese cabbage kimchi added with 7% red pepper powder and 2.8% garlic (K-2), 7% red pepper powder, 2.8% garlic, Chinese cabbage kimchi with 0.1% chopi powder, 7 Four kinds of Chinese cabbage kimchi (K-4) added with 4% red pepper powder, 2.8% garlic, 0.1% chopi powder and 5% freshly prepared cabbage kimchi and administered to mice. The NO production predicted to be produced by acting bacterial toxins was compared. Intraperitoneal macrophages were isolated from the experimental groups 3 weeks after the administration of 0.5 mg of kimchi each day into the mouth of each mouse. Nitric oxide (NO), which is expressed by the action of lipopolysacchride (LPS, 10 μg) on isolated macrophages, was measured. As a control group, only saline was administered. As a result, as shown in Figure 1, the standardized Chinese cabbage kimchi administration group (K-1 and K-2) was slightly less NO production than the control group, but increased the amount of red pepper powder and garlic, added crust (K-3), gat and chopi (K-4) added organic high activity cabbage kimchi group showed an effect to significantly increase the NO production.

Example 10 Preparation of Anti-cancer Functionality Organic High Activity Chinese Cabbage Kimchi

In this example, according to the results obtained in Examples 1 to 9, according to the recipe of anti-cancer functional enhancement cabbage kimchi that can obtain the best anti-cancer effect, Chinese cabbage kimchi was prepared, and cancer prevention and anti-cancer promotion effects were investigated in vivo. . That is, using the same standardized cabbage kimchi (CⅠ) and standardized cabbage kimchi (OCI) using the material prepared by the organic method, the same as in Example 1, based on the standardized cabbage kimchi according to the results obtained in Examples 1 to 9 As the cabbage kimchi of general cabbage high active cabbage kimchi (CII) added with red pepper powder, garlic, chopi, fresh water 7%, 2.8%, 0.1%, 5%, respectively, and the results obtained in Examples 1-9 above Based on the standardized Chinese cabbage kimchi prepared organic high activity cabbage kimchi (OCII) added with red pepper powder, garlic, chopi, freshly prepared 7%, 2.8%, 0.1%, 5% to the organic cabbage, respectively, fermented at 15 ℃ for 1 day After fermentation at 5 ° C. until the pH was 4.3, lyophilization was performed to produce a methanol extract. This methanol extract was prepared as a sample to examine the effect of life extension and inhibition of solid cancer growth after transplantation of sarcoma-180 tumor cells into Balb / c mice in vitro and antimutagenic and anticancer experiments.

Example 11 Anticancer Effect of Organic High Activity Chinese Cabbage Kimchi by MTT Method

In the present embodiment, the seasoning ratio (Ⅰ) and the standardized seasoning ratio of the standardized Chinese cabbage kimchi prepared in Example 6 (II) to improve the functionality of general cabbage (CⅠ) kimchi, organic cabbage (OCⅠ) kimchi, The anticancer effect of the anticancer functional organic high activity cabbage kimchi of the present invention was tested by the same method as the MTT assay performed in Example 3 with general high activity cabbage (CII) kimchi and organic high activity cabbage (OCII) kimchi. As a result, as shown in Table 24, the inhibitory effect of AGS human gastric cancer cells in the MTT assay showed that the normal cabbage (CI) kimchi showed 61% and 67% at 5mg / ml and 10mg / ml, respectively. Chinese cabbage (CII) kimchi was 73% and 72%, while organic cabbage (OCII) kimchi was 78% and 83%. By Chinese cabbage, organic cabbage showed higher inhibition rate than general cabbage, and the higher ratio of marinated fermentation (II) than standardized seasoning (Ⅰ).

Survival Inhibitory Effect of Cabbage Kimchi Methanol Extract on AGS Human Gastric Cancer Cells by MTT Method Treated product (mg / ml) OD ₅₄₀ 5 10 Control 0.873 ± 0.128 0.680 ± 0.044 C ¹⁾ (Ⅰ) Kimchi 0.511 ± 0.014 (61) ³⁾ 0.443 ± 0.031 (67) OC ²⁾ (Ⅰ) Kimchi 0.470 ± 0.083 (66) 0.439 ± 0.074 (71) C (Ⅱ) Kimchi 0.493 ± 0.093 (73) 0.457 ± 0.022 (72) OC (Ⅱ) Kimchi 0.407 ± 0.148 (78) 0.354 ± 0.016 (83) 1) Regular Chinese cabbage 2) Organic Chinese cabbage 3)

Example 12: anticancer effect and immune activity enhancement effect of high activity cabbage kimchi in vivo

In this example, the anticancer effect and the immune activity enhancement effect of high activity cabbage kimchi in vivo was investigated.

Balb / c mouse, an experimental animal used in this example, was used as the weight of about 25g and feed was raised as a standard feed. During breeding, water and feed were supplied in sufficient quantities and the animal room maintained a light-dark cycle of 12 hours.

Tumor cells used in this Example were taken with a plurality of sacroma-180 cells incubated for one week in the abdominal cavity of the experimental animal and centrifuged with phosphate buffered saline (PBS) (1,200rpm, 10min) to separate the tumor cells. The separated cells were suspended in PBS again, centrifuged again to remove the supernatant, and then sacroma-180 cells were made in a 1 × 10 ⁶ cells / mL tumor cell suspension to be intraperitoneally injected and used for experiments.

The sample used in this example was prepared using sterilized PBS with methanol extracts of general cabbage (CⅠ) kimchi, organic cabbage (OCⅠ) kimchi, general high activity cabbage (CII) kimchi, and organic high activity cabbage (OCII) kimchi. The dose was 0.5 mg per mouse, and the control group was injected with sterile PBS only and was stored while stored in the refrigerator when not administered.

Experimental Example 1: Viability test

In order to investigate the direct cytotoxicity of kimchi samples, viability test was performed in vitro using dye exclusion method. 24-well plates were seeded at 5 × 10 ⁴ cells / plate in 1 mL of Eagle's minimal essential medium (EMEM) medium containing 20% HFCS (heat inactivated fetal calf serum: Gibco Lab, USA). 20 μl of Kimchi sample was added thereto and incubated in 37 ° C., 5% CO ₂ , and incubator for 24 hours. After 24 hours of incubation, 50 μl of cells were mixed well with 50 μl of 0.2% trypan blue solution, and the total number of cells, the number of non-viable cells and the number of viable cells were measured using a hemocytometer. After measurement, the viability was calculated according to the following formula compared with the control cell group without the kimchi sample.

As a result, as shown in Table 25, the cytotoxic effect of the sample methanol extract on sarcoma-180 mouse tumor cells was not affected at 0.75 mg / mL.

Cytotoxicity Test of Methanol Extract of Organic Highly Active Chinese Cabbage Kimchi Sample Final concentration (mg / ml) Total cell number (× 10 ⁴ ) Saint John's Cells (%) Control 17.4 ± 2.9 92 S-180 + C (Ⅰ) Kimchi 0.1 12.8 ± 2.1 85 0.5 11.8 ± 3.1 80 0.75 10.4 ± 3.0 77 1.0 7.2 ± 3.9 62 2.5 7.0 ± 2.4 64 S-180 + OC (Ⅰ) Kimchi 0.1 11.8 ± 3.4 84 0.5 10.2 ± 2.6 77 0.75 10.8 ± 2.8 75 1.0 7.5 ± 0.9 66 2.5 5.6 ± 2.9 56 S-180 + C (Ⅱ) Kimchi 0.1 11.6 ± 4.6 88 0.5 10.2 ± 2.4 85 0.75 8.4 ± 2.9 81 1.0 6.2 ± 1.3 71 2.5 6.8 ± 1.9 59 S-180 + OC (Ⅱ) Kimchi 0.1 13.0 ± 2.1 83 0.5 10.2 ± 2.3 75 0.75 12.4 ± 2.1 81 1.0 8.4 ± 2.2 64 2.5 7.2 ± 2.5 58

Experimental Example 2: Life Extension Effect

Seven animals per group were injected into the peritoneal cavity of 1 mL (1 × 10 ⁶ cells / mouse) of tumor cell suspension prepared by the method described above, and then the samples were intraperitoneally administered for 24 days to 20 days after 24 hours. We observed the survival rate of and calculated the lifespan days. As a result, as shown in Table 26, the control survived 27 days, whereas the normal cabbage high activity (CII) cabbage kimchi showed the highest life extension effect of 32% (39.7 days) and the organic cabbage (OCⅠ) cabbage kimchi and The organic high activity cabbage (OCII) cabbage kimchi showed a high life extension effect of 26% with 36.5 days.

Effect of Methanol Extract of Organic Highly Active Chinese Cabbage Kimchi on the Life Span of Balb / c Mice Sample Survival Time (day) Extension rate (%) Control 27.0 ± 1.7 - S-180 + C (Ⅰ) Kimchi 35.2 ± 1.7 23 S-180 + OC (Ⅰ) Kimchi 36.5 ± 2.9 26 S-180 + C (Ⅱ) Kimchi 39.7 ± 2.9 32 S-180 + OC (Ⅱ) Kimchi 36.5 ± 2.8 26

Experimental Example 3: Solid Cancer Growth Retardation Effect

Six animals from each group were subcutaneously implanted with 0.2 mL (6 × 10 ⁶ cells / mouse) of tumor cell suspension in the laboratory at weekly intervals in the laboratory for 20 hours. The sample solution was administered intraperitoneally once daily. On the 26th day of tumor cell transplantation, the solid tumors generated by lethality were extracted and weighed, and the tumor growth inhibition ratio (IR:%) was calculated according to the following equation.

Cw: mean tumor weight of control group

Tw: mean tumor weight of treatment group

As a result, as shown in Table 27, the growth inhibition effect of solid cancer was high at 34.3%, 29.5%, and 39.3%, respectively. Of the four kimchi varieties, Kimchi, which was developed with high organic activity, was most effective in inhibiting solid cancer growth.

Inhibitory Effects of Chinese Cabbage Kimchi Extracts on Solid Carcinoma Growth in Balb / c Mice with Sacroma-180 Cells Sample Tumor Weight (g) Inhibition Ratio (%) S-180 + PBS 5.19 ± 1.19 - S-180 + C (Ⅰ) Kimchi 4.57 ± 1.41 11.9 S-180 + OC (Ⅰ) Kimchi 3.41 ± 1.09 34.3 S-180 + C (Ⅱ) Kimchi 3.66 ± 0.73 29.5 S-180 + OC (Ⅱ) Kimchi 3.15 ± 0.65 39.3

Experimental Example 4: Weight change of each organ of mouse

In Experimental Example 3, the weights of the organs of the Balb / c mouse that completed the solid cancer growth inhibition experiment were weighed. As a result, as shown in Table 28, the weight per spleen of organic cabbage (Ⅰ) kimchi was slightly higher, but the weight per weight of other organs was similar.

Balb / c Mouse Weight of Each Organ Sample Weight (g) Spleen / weight wt. (%) Liver / weight wt. (%) Heart / weight wt. (%) Height / weight wt. (%) Control 29.0 ± 1.63 0.47 ± 0.13 8.68 ± 1.15 0.65 ± 0.09 2.16 ± 0.19 S-180 + PBS 30.2 ± 1.00 1.50 ± 0.17 7.78 ± 0.56 0.55 ± 0.04 1.78 ± 0.06 S-180 + C (Ⅰ) Kimchi 26.6 ± 1.62 2.35 ± 0.3 8.77 ± 0.62 0.68 ± 0.12 2.39 ± 0.33 S-180 + OC (Ⅰ) Kimchi 28.9 ± 1.23 1.39 ± 0.17 6.93 ± 0.56 0.74 ± 0.11 1.76 ± 0.09 S-180 + C (Ⅱ) Kimchi 28.1 ± 0.90 1.82 ± 0.21 8.07 ± 0.91 0.72 ± 0.11 1.92 ± 0.33 S-180 + OC (Ⅱ) Kimchi 27.2 ± 2.07 1.77 ± 0.16 8.38 ± 0.96 0.61 ± 0.18 2.01 ± 0.33

Experimental Example 5: Changes in glutathione-S-transferase (GST) activity and content of glutathione (GSH) and lipid peroxide (LPO) in mouse liver

Glutathione-S-transferase (GST), known as an anti-carcinogenic enzyme, is a detoxifying agent of active metabolites of electrophilic carcinogens. Although activated by enzymes to cause DNA damage, GST acts as a substrate by using an active metabolite having these hydrophobic residues as a substrate, and covalently binds GSH to a stabilized form, and finally detoxifies it by excretion in urine.

In this experimental example, mice were first killed to measure glutathione-S-transferase (GST) activity, and then liver was perfused with physiological saline of 4 ° C. or lower to remove blood remaining in the liver, and the liver was extracted. 0.1 M potassium phosphate buffer (pH 7.4) was added per 1 g of liver tissue, and the mixture was ground with a glass teflon omogenizer under ice cooling. This grinding solution was used as a homogenate fraction, which was centrifuged at 13,000 rpm for 10 minutes to remove the nuclei and unbranched cell sections, and the supernatant obtained by ultracentrifugation for 1 hour at 105,000 × g was used as the cytosol fraction. It was. The homogenate fraction thus obtained was used to measure glutathione content and lipid peroxide content, and the cytosol fraction was used to measure glutathione S-transferase activity. According to the method of Habig et al., 75 μl of 0.04 M reduced glutathione in 0.1 M potassium phosphate buffer (pH 6.5) was added, 0.1 mL of enzyme solution was added, 0.5 mL of 20% trichloroacetic acid was added to the blank, and the sample was inactivated at 25 ° C. After reacting for 5 minutes, add 25 µl of 0.12M 1-chloro-2,4-dinitrobenzene to the blank and each sample, and react for 2 minutes at 25 ° C. Then, add 20% trichloroacetic acid to the sample to complete the reaction. The supernatant obtained from the separation was measured for absorbance at 340 nm, and then the activity was calculated using the molar extinction coefficient of 1-chloro-2,4-dinitrobenzene at 9.6 mM ¹ cm ^-1 . Units of enzymatic activity were expressed as nmole number of 2,4-dinitrobenzene-glutathione produced by mg protein for 1 minute. In addition, the content of glutathione in the tissue was determined by dissolving the supernatant in which the protein was removed by adding 4% sulfosalicylic acid, an alkaloid reagent, to the enzyme source (400-600 µg protein) according to Ellaman's method. 2.7 mL of 5,5'-dithiobis (2-nitrobenzoic acid) was added to 0.1 M potassium phosphate buffer (pH 8.0), and the resulting blue color was measured at 412 nm and calculated according to a standard curve. It is expressed as glucose glutathione μmole. As shown in FIG. 2, the changes of GST activity in liver tissues were 680nmol / mg protein / min and 747nmol / mg for organic cabbage (OCI), general high activity cabbage (CII) kimchi and organic high activity cabbage (OCII) kimchi, respectively. protein / min, 726 nmol / mg protein / min was high (p <0.05). Glutathione content was shown in organic cabbage (OCI) kimchi, general high activity cabbage (CII) kimchi, organic high activity cabbage ( OCII) Kimchi was high as 0.40μmol / g, 0.38μmol / g and 0.43μmol / g, respectively. Lipid peroxide, a lipid that has been added to oxygen by adding oxygen to unsaturated fatty acids, is a lipid component, and unsaturated fatty acids that form phospholipids, which are the major components of cell membranes, are produced by binding to active oxygen. As shown in FIG. 4, the lipid oxide content of the general high activity cabbage (CII) kimchi and the organic high activity cabbage (OCII) kimchi were 6.7 nmol / g and 2.6 nmol / g.

Experimental Example 6: Analysis of natural killer cell activity

Natural killer (NK) cells are monocytes that are naturally killer and are non-detoxifying and non-adherent cells that selectively destroy lymphoid cells expressing RNA tumor virus antigens. It is recognized as an important immune function involved in tumor resistance as a different immune surveillance apparatus. Therefore, in the present experimental example, NK cells, which are effector cells, were prepared in order to analyze the activity of natural killer cells. The spleens of Balb / c mice were aseptically extracted on a clean bench, washed three times with 5 mL of RPMI 1640 medium containing penicillin (100 U / mL) and streptomycin (100 μg / mL), Crushed. The cell suspension was filtered through a 70 μm nylon mesh, centrifuged to collect lymphocytes, and then suspended in the same medium. Lymphocytes were isolated by centrifugation (500 × g, 30 min, 18 ° C.) using histopacque-1077 (histopaque-1077). The prepared cells were resuspended in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine, penicillin (100 U / mL), streptomycin (100 μg / mL). This was incubated for 1 to 2 hours at 37 ° C. in a CO ₂ incubator to allow cells to adhere to the culture flask (6 cm Petri dish). Non-adhesive natural killer cells were collected by centrifugation, resuspended in culture medium, and the appropriate number of cells was counted. Subsequently, cell destruction activity of natural killer cells was analyzed by MTT method. Yac-1 mouse lymphoma cells were used as target cells. Yac-1 cells are the most convenient target cell line for measuring NK cell activity. Yac-1 cells were diluted to a density of 5 × 10 ⁴ cells / mL in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine phenylsilin (100 U / mL) and streptomycin (100 μg / mL). 50 μl was added to each well. In addition, NK cells were added to the 96 well plate, 50 μl of effector cells and Yac-1 cells, respectively, for NK cell activity analysis of 1 × 10 ⁷ cells / mL. After incubating for 3 days at 37 ° C. in a CO ₂ incubator, 10 μl of MTT (5 mg / mL) solution was added to each well and incubated at 37 ° C. for 4 hours. 25 μl of SDS (10% in 0.02N HCl) was added thereto, followed by color development at room temperature for 30 minutes, and OD was measured at 540 nm. Percent cytotoxicity was calculated by the following formula.

As shown in FIG. 5, NK cells of mice treated with organic cabbage (OCI) kimchi, general cabbage (CII) kimchi and organic cabbage kimchi (OCII), compared to 18.5% of the control (S-180 + PBS) as shown in FIG. 46%, 44%, and 79% of the cells showed high cytotoxic activity, respectively.

As shown in Experimental Examples 1 to 6, as the cabbage type, organic cabbage had an anticancer effect than a general cabbage, and the spice ratio in vivo anticancer in kimchi (II) kimchi enhanced functionality rather than standardized spice ratio (I). The effect was enhanced.

Example 13 Antitumor and Immune Activity Enhancement Effects of Organic High Activity Chinese Cabbage Kimchi Diet

In this embodiment, the kimchi diet containing organic high-activity cabbage kimchi and general high-activity cabbage kimchi, respectively, was prepared, and the part of the cabbage kimchi as well as the cabbage kimchi itself was investigated by investigating their inhibitory effect on the growth of solid cancer and the cytotoxic activity of natural killer cells. The anticancer function of the diet containing was also investigated. Kimchi diet was prepared as shown in Table 30 on the basis of the general component analysis shown in Table 29 and the experimental method was carried out in the same manner as in Example 13, and the standardized Chinese cabbage kimchi, general high activity cabbage kimchi, organic high activity cabbage kimchi Was fed with a diet made by adding 4 and 8%, respectively, for 2 weeks and injected with sarcoma-180 cells in the inguinal part of the mouse for 30 days. As a result, as shown in Table 31, the inhibition rate of the solid cancer inhibition effect was the lowest with the standardized 4% added cabbage kimchi 3% and the highest with the organic high activity 8% cabbage kimchi 28%. In generalized cabbage kimchi, 8% added cabbage kimchi showed higher inhibition rate than 4%, and general high activity cabbage kimchi was slightly higher than 8% added cabbage kimchi, but there was no significant difference. Chinese cabbage kimchi added 8% higher than 4%. According to the group of Chinese cabbage kimchi, the general high activity cabbage kimchi was higher than the general standardized cabbage kimchi and the organic high activity cabbage kimchi showed the highest inhibition rate. In addition, as shown in FIG. 6, the activity of the natural killer cells that act on the immune system was higher than 4% of generalized cabbage kimchi and 8% of Chinese cabbage kimchi added. Kimchi had 8% more activity than Chinese cabbage added 4%. According to the group of Chinese cabbage kimchi, the diet with generalized cabbage kimchi was similarly lower than the control group, and the diet with general high activity cabbage kimchi had higher activity than the diet with generalized cabbage kimchi. NK cells showed the highest immune activity when fed diets containing organic high activity cabbage kimchi.

Table of General Components of Kimchi Prepared General standardization General high activity Organic high activity moisture 4.86 5.59 3.79 Fat 2.87 3.09 4.71 protein 15.76 15.42 13.53 Sugar 15.12 12.16 12.16 fibrin 39.64 43.39 48.04 Ash 21.75 20.35 17.77

Chinese cabbage kimchi dietary composition Standard diet 4% of schedules 8% of schedules 4% 8% 4% 8% Casein 20.0 19.37 18.77 19.38 18.77 19.46 18.92 _DL -Methionine 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Corn starch 15.0 15.0 15.0 15.0 15.0 15.0 15.0 Sucrose 50.0 49.4 48.79 49.51 49.03 49.51 49.03 Fibrous 5.0 3.41 1.83 3.26 1.53 3.08 1.16 Corn oil 5.0 4.89 4.77 4.88 4.75 4.81 4.62 Mineral mixtures 3.5 3.5 3.5 3.5 3.5 3.5 3.5 Vitamin mixtures 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Choline Bitrate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Standard diet: Normal diet (AIN diet) 4%: Standard diet + general standardized kimchi 4% added day 8%: Standard diet + general standardized kimchi 8% added 4%: Standard diet + general high activity kimchi 4% added 8% : Standard diet + high activity kimchi 8% added 4%: Standard diet + organic high activity kimchi 4% added 8%: standard diet + organic high activity kimchi 8% added

Solid rock growth inhibition activity Diet Tumor weight (g) Inhibition Ratio (%) S-180 + Control 6.0 ± 1.3 - S-180 + Day 4% 5.8 ± 1.0 3 8% of S-180 + schedule 4.9 ± 1.0 18 4% S-180 + 4.9 ± 0.9 18 8% S-180 + 5.1 ± 0.7 15 S-180 + note 4% 4.8 ± 0.9 20 S-180 + note 8% 4.3 ± 1.0 28

Example 14 Cancer Promoting Effect of Highly Active Chinese Cabbage Kimchi

In this Example, superorganisms using reticulocytes of peripheral blood of mouse in order to confirm the cancer prevention effect of the organic high activity cabbage kimchi prepared according to the recipe of the best anti-cancer functional cabbage kimchi prepared in Example 6 in vivo A supravital staining micronucleus assay was performed. Micronucleus assays using rodent bone marrow polychromatic erythocytes (PCE) are cytogenic assays for the induction of micronuclei, an abnormal chromosomal component formed during the differentiation of red blood cells.

6-7 weeks old male ICR mice were injected intraperitoneally to dissolve mitomycin (MIC) in physiological saline as positive control. The dose of the test substance is intraperitoneally injected at a maximum value of 1/2 of the 50% lethal dose (LD ₅₀ ). Blood was collected at intervals. 5 μl of blood was taken from the tail blood vessels of the intraperitoneally injected mice, dropped onto an acridine orange-coated slide, covered with a cover glass, and left for 4 hours at 4 ° C. Allow acridine orange to react sufficiently. The slides were observed at 10 × 100 times by fluorescence microscopy. Reticulocytes were counted from type I to type III to about 2000. Among them, reticulocytes with micronuclei were counted and statistically evaluated by Cochran Armitage method. As a result, as shown in Figure 7, oral administration of 500mg / kg standardized cabbage kimchi showed no micronucleus-induced inhibitory ability, but the organic high activity cabbage kimchi showing the best cancer prevention function significantly (p <0.05) micronucleus Induced inhibitory effect. In addition, in the group administered orally at a dose of 1000 mg / kg, cancer preventive functional kimchi has a significantly higher effect of inhibiting micronucleus induction than standard cabbage kimchi (p <0.05). Has been shown to improve cancer prevention.

Example 15: Tumor Metastasis Inhibitory Effect of Organic High Activity Chinese Cabbage Kimchi

Animals used in this experiment, Balb / c mice (female, 6 weeks old) were purchased from the Korea Research Institute of Chemical Technology (Daejeon), and feed was raised as standard feed. During breeding, water and feed were supplied in sufficient quantities. The animal laboratory maintained a temperature of 22 ± 1 ℃, a humidity of 55 ± 5%, and a light-dark cycle at 12 hour intervals. To measure the tumor metastasis suppression effect of Chinese cabbage kimchi, colon 26-M3.1 carcinoma (Dr. Yoon Taek-Joon, Handong University, Ph.D.), a metastatic tumor cell with metastasis to the lung, was cultured in virto (7.5% FBS containing 100 μl (2.5 × 10 ⁴ mL) of the culture solution was intravenously injected into the tail of Balb / c mice. Each group consisted of 5 rats, and the Chinese cabbage kimchi extract was injected subcutaneously 2 days before tumor cell inoculation. The metastasis of the tumor was determined 14 days after vaccination, the mouse was sacrificed to remove the lung, the target organ of the tumor, and then the tumor was colonized after fixing the tumor in Bouin's solution (saturated picric acid 15: formaline 5: acetic acid 1). The number was counted and the effect of inhibiting tumor metastasis by Chinese cabbage kimchi was measured by comparing with the tumor-inoculated control group. As a result of the experiment, as shown in Table 32, the experimental results of the prophylactic effect of tumor metastasis using colon 26-M3.1 carcinoma showed 14% tumor metastasis suppression when subcutaneous injection of 1.25mg / mouse in the standardized cabbage kimchi I group. It showed an effect. Organic high activity cabbage kimchi showed a 12% metastasis inhibitory effect similar to that of the standardized cabbage kimchi I group at a low concentration of 0.05 mg. When subcutaneous injection of 1.25 mg, the growth of colon 26-M3.1 tumors that metastasized to the lung was 35%. It was confirmed that the organic high activity cabbage kimchi also had a preventive effect of cancer metastasis.

Tumor Metastasis Inhibitory Effect of Highly Active Chinese Cabbage Kimchi Using Colon 26-M3.1 Cells Treatment Dose (mg / mouse) root Lung cancer metastasis (% inhibition) Mean ± SD range Control sc 162 ± 7 ^a 153 ~ 172 C (I) Kimchi 0.050.251.25 scscsc 157 ± 13 ^ab (3) ¹⁾ 147 ± 8 ^ab (9) 139 ± 5 ^bc (14) 142 ~ 172138 ~ 157131 ~ 144 OC (Ⅱ) Kimchi 0.050.251.25 scscsc 143 ± 12 ^b (12) 125 ± 15 ^c (23) 103 ± 5 ^d (36) 124 ~ 156109 ~ 14696 ~ 108 ¹⁾ Values in parentheses indicate significant differences (p <0.05) in percent inhibition ^{a to d} Duncan's multiple range test.

As described above through the above Examples and Experimental Examples

For pickled cabbage 100, we have recipes of radish 13.0%, green onion 2.0%, red pepper powder 7%, garlic 2.8%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0%, final salt 2.5%, chopi 0.1%, fresh 5% The anticancer functional cabbage kimchi of the present invention and the diet containing the same, which have been fermented again at a low temperature of 5 ° C. until the primary fermentation at 15 ° C. and pH 4.3, have an excellent effect of showing anticancer function not only in vitro but also in vivo. Kimchi is also a very useful invention in the food industry because it has an excellent effect of increasing the preservation of kimchi.

Claims (6)

It contains 100% pickled cabbage, radish 13.0%, green onion 2.0%, red pepper powder 7%, garlic 2.8%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0%, final salt 2.5%, chopi 0.1%, fresh 5% Anti-cancer functional cabbage kimchi characterized by. Contains 100% pickled cabbage, radish 13.0%, green onion 2.0%, red pepper powder 7%, garlic 2.8%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0%, final salt 2.5%, chopi 0.1%, fresh 5% And fermentation at 15 ° C. and fermentation again at 5 ° C. until the pH is 4.3. The anti-cancer functional cabbage kimchi according to claim 1 or 2, wherein the Chinese cabbage, radish, green onion, ginger, red pepper powder, garlic, chopi, and freshly grown organically. The anti-cancer functional cabbage kimchi according to claim 1 or 2, wherein the component of the final salt selects one or more of processed salts consisting of roasted salt, fresh salt, and bamboo salt. For pickled cabbage 100, mix 13.0% radish, 2.0% green onion, 7% red pepper powder, garlic 2.8%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0%, final salt 2.5%, chopi 0.1%, fresh 5% And fermentation at 15 ° C. and fermentation again at 5 ° C. until it becomes pH 4.3. Chinese cabbage kimchi according to claim 1, 4-8%, casein 18-20, DL-methionine 11%, corn starch 15%, sucrose 49-50%, fiber 1-3, corn oil 4-5%, minerals Anti-cancer functional cabbage kimchi diet composition consisting of 3.5% mixture, 1.0% vitamin mixture.