KR100364430B1 - Anticancer activities of chinese cabbage kimchi containing KCl and process for preparation thereof - Google Patents

Abstract

The present invention relates to anti-cancer cabbage kimchi containing KCl and a method of manufacturing the same. For pickled cabbage 100, radish 13.0%, green onion 2.0%, red pepper powder 3.5%, garlic 1.4%, ginger 0.6%, anchovy sake 2.2%, sugar 1.0 %, Standardized cabbage kimchi or pickled cabbage containing 2.5% final salt 7% red pepper powder, garlic 2.8%, chopi 0.1%, fresh 5%, ginger 0.6%, radish 13.0%, green onion 2.0%, anchovy sauce 2.2% Enhancement of cancer prevention function, containing 1.0% of sugar and 2.5% of final salt By replacing 20-40% of the salt added to kimchi with KCl, it has an excellent effect of enhancing anticancer activity without changing the physicochemical and sensory properties of Chinese cabbage kimchi. There is.

Description

Anti-cancer cabbage kimchi containing BCC and its manufacturing method {Anticancer activities of chinese cabbage kimchi containing KCl and process for preparation}

The present invention relates to anti-cancer cabbage kimchi containing KCl and a method for producing the same. More specifically, the present invention relates to an anticancer cabbage kimchi and a method for producing the cabbage kimchi, which has been replaced with a portion of salt in the production of cabbage kimchi to enhance the anticancer properties of the cabbage kimchi.

Kimchi is an indispensable food in our diet, and much research has recently been conducted on its functionality. 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 (Korean Patent Publication -13238), manufacturing method of cabbage kimchi added ginseng (Korean Patent Publication No. 96-13237), manufacturing method of ginseng Kimchi (Korean Patent Publication No. 98-025327), manufacturing method of seaweed kimchi (Korean Patent Publication No. 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. However, until now, researches aiming to obtain the effect by simply adding ingredients having anticancer effect to kimchi have been carried out.

On the other hand, the cause of gastric cancer in the stomach cancer of our country is caused by salted food, especially the consumption of salty and spicy food is dominant, Kimchi can be representative salty and spicy food. Therefore, efforts have been made to minimize the amount of salt added to the kimchi immersion, but if the salt content is reduced too much, there is a disadvantage that the storage and texture of the kimchi becomes worse.

The present inventors studied to reduce the amount of salt related to cancer occurrence and have no damage to storage and texture, but rather to produce kimchi with increased anticancer activity, and when replacing some of the salt added during the production of kimchi with KCl, The present invention was completed by confirming that anticancer activity is greatly increased without changing storage properties and texture.

Accordingly, an object of the present invention relates to an anticancer cabbage kimchi containing KCl prepared by replacing some salts added to cabbage kimchi with some KCl. Another object of the present invention to provide a method for producing anti-cancer cabbage kimchi containing the KCl.

The purpose of the present invention is to compare the peroxidation promoting effect, mutagenicity, gastric cancer cell growth inhibitory effect of NaCl and KCl itself, and the cytotoxicity to 3-methyl cholanthracene (MCA) in C3H10T1 / 2 After examining the inhibitory and anti-carcinogenic effects, it was confirmed that KCl-containing cabbage kimchi was superior to NaCl, and the antimutagenic effect of KCl-containing cabbage kimchi and its anti-cancer effects and immunity in vitro and in vivo were investigated. Investigation of the activity enhancing effect confirmed that the present invention KCl-containing cabbage kimchi is anti-cancer and subcutaneous injection of methanol extract of KCl-containing cabbage kimchi in the mouse and then injected into the tail of a tumor cell line with metastasis in the lung This was achieved by investigating the effects of tumor metastasis.

Hereinafter, the configuration and operation of the present invention.

Figure 1 shows the effect of NaCl and KCl on the automatic oxidation of linoleic acid.

Figure 2 shows the pH change during the fermentation period of the cabbage kimchi of the present invention containing KCl and the standardized cabbage kimchi and cancer prevention functional enhanced cabbage kimchi.

Figure 3 shows the change in acidity during the fermentation period of the cabbage kimchi of the present invention containing KCl and standardized cabbage kimchi and cancer prevention functional cabbage kimchi.

Figure 4 is Leuconostoc sp. During the fermentation of the Chinese cabbage kimchi and the standardized Chinese cabbage kimchi and cancer prevention functional enhancement cabbage kimchi containing KCl. Indicates a change of.

Figure 5 is the Lactobacillus sp. During the fermentation of the Chinese cabbage kimchi and the standardized Chinese cabbage kimchi and cancer prevention functional enhancement cabbage kimchi containing KCl. Indicates a change.

FIG. 6 shows QDA (Sensory Test) results of cancer preventive functional enhanced kimchi fermented at 15 ° C. for 5 weeks at 5 ° C. and standardized cabbage kimchi fermented at 15 ° C. for 1 week at 5 ° C. for 3 weeks.

7 is a liver cytosolic glutathione-S-transferase after transplanting sarcoma-180 cells to Balb / c mice and administering the cabbage kimchi of the present invention containing KCl and the standardized cabbage kimchi and cancer prevention functional cabbage kimchi methanol extract The result of measuring the activity of is shown.

Figure 8 shows the effect of the present invention cabbage kimchi containing the KCl and the standardized Chinese cabbage kimchi and cancer prevention functional enhanced Chinese cabbage kimchi methanol extract on the natural killer cell activity of mouse spleen lymphocytes.

Figure 9 is a photograph showing the tumor metastasis inhibitory effect of colon 26-M3.1 cells treated with methanol extract of the control and cancer prevention functional enhanced kimchi II (Gueun salt + KCl).

The present invention comprises the steps of adding NaCl and KCl to linoleic acid at a concentration of 0.2, 0.4, 0.6% to autooxidize and then measure the peroxide value (POV) to determine that NaCl significantly increases the POV than KCl; NaCl and KCl treated with Salmonella typhimurium TA100 strain with MNNG at a concentration of 3, 7, 15, 25% and counting the return mutation number to demonstrate that the mutation of KCl is superior to NaCl; MCA was added to cultured C3H / 10T1 / 2 cells and then reinoculated with medium containing FCS (fetal calf serum) and cultured to count the colony of colonized cells, thereby inhibiting cytotoxicity of MCA. Investigating the effect and culturing C3H / 10T1 / 2 cells, and then re-incubated with MCA, NaCl, and KCl, and then counting the transformed foci to investigate the anti-carcinogenic effect; Producing six kinds of Chinese cabbage kimchi by replacing 30% of the salt used for preparing standardized Chinese cabbage kimchi and cancer prevention functional kimchi with KCl; Six kinds of cabbage kimchi prepared above were fermented at 5 ° C. for 6 weeks while measuring pH change and acidity over time, and involved in taste, Leuconostoc sp. Lactobacillus sp. Is involved in the production of lactic acid bacteria and acids. Investigating the physicochemical and organoleptic properties of the six cabbage kimchi prepared by measuring the amount of lactic acid bacteria; Investigating the antimutagenic effect of the methanol extracts of the six kinds of Chinese cabbage kimchi against the AFB ₁ of Salmonella typhimurium TA100 strain and the inhibitory effect of MCA on C3H / 10T1 / 2 cells; AGS human stomach cancer cells were incubated with the methanol extracts of the six kinds of Chinese cabbage kimchi, and then cultured with MTT reagent to investigate the effect of inhibiting cancer cell proliferation. Examining the effect of inhibiting cancer cell growth by measuring the number of cells; Among the six kinds of Chinese cabbage kimchi, methanol extracts obtained from four kinds of Chinese cabbage kimchi including KCl including KC Kimchi were added to Sarcoma-180 cells for cultivation, followed by cultivation to show direct toxicity to Sarcoma-18 cells. Finding a concentration of the sample; 10 mice were subcutaneously transplanted with tumor cells Sarcoma-180 and mice were intraperitoneally administered with a concentration of methanol extract samples obtained from four kinds of Chinese cabbage kimchi including KCl, including standardized cabbage kimchi that did not exhibit direct cytotoxicity. Measuring change and weight of solid rock; Anti-carcinogenicity by measuring the activity change of glutathione-S-transferase (GST) enzymes involved in the transfer and excretion of toxic substances and peroxides in the liver of glutathione (Glutathione) in the liver of the mouse Investigating; Yac-1 cells were used as target cells and NK cells prepared from lymphocytes isolated from the spleen of Balb / c mice were used as effector cells to investigate the immune enhancing effect of natural killer cells by MTT assay; Two days before intravenous tumor cell injection, methanol extracts from four cabbage kimchi containing KCl, including standardized Chinese cabbage kimchi, were injected intravenously with colon 26-M3.1 carcinoma, a classical tumor cell line, in the tail of Balb / c mice. Injecting subcutaneously 14 days after the date of tumor cell inoculation to investigate the effect of inhibiting tumor metastasis.

In the present invention, the standardized Chinese cabbage kimchi is the standardized Chinese cabbage kimchi recipe through the sensory test and the physicochemical test according to the material mixing ratio of the cabbage kimchi, and in June 1998, the Korean Journal of Food Science (HSKCAN 39 (2)) The ideal cabbage kimchi composition ratio, published in a standardized study of 1,3% radish, 2.0%, red pepper powder 3.5%, garlic 1.4%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0%, final salt 2.5 Chinese cabbage kimchi containing%.

In the present invention, the cancer preventive function-promoting kimchi was determined by the present inventors by modifying the ratio of the subsidiary material to the standardized cabbage kimchi to determine the best anti-cancer cabbage kimchi recipe, pepper powder 7%, garlic 2.8%, garlic 0.1% for pickled cabbage 100 Cancer preventive functional enhancement kimchi containing 5% freshly, 0.6% ginger, 13.0% radish, 2.0% green onion, 2.2% anchovy, 2.2% sugar, and 2.5% final salt. It includes.

The KCl-containing cabbage kimchi of the present invention, the amount of KCl contained 20% to 40% based on the total salt content, preferably 30%.

Human gastric cancer cells (AGS hunan adenocarcinoma cells) among the cell lines used in the present invention were distributed from the Bank of Korea Cell Line (Seoul National University of Medicine) and Sarcoma-180 cells 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 an animal Balb / c mouse (female, 6 weeks old) was purchased from the Korea Research Institute of Chemical Technology in Daejeon.

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 Comparison of Biological Activities of NaCl and KCl

Peroxide value (POV), mutation and gastric cancer cell growth inhibitory effects were measured in relation to cancer.

Experimental Example 1: Comparison of the peroxidation promoting effect of NaCl and KCl itself

After injecting 1.00 g of linoleic acid into the Erlenmeyer flask (100 mL), 20 mL of ethanol was added, dissolved, and 25 mL of phosphate buffer (0.2M, pH7.0) was added and mixed. In this case, NaCl and KCl, which were added at this time, were dissolved in ethanol and phosphate buffer at 0.2, 0.4, and 0.6% concentrations, respectively, and the corresponding concentrations were added to the Erlenmeyer flask. After automatic oxidation at 50 ° C. for the reaction time, these reaction solutions were transferred to a 300 mL separatory funnel, and then extracted three times using a small amount of water and 25 mL of chloroform. The lower layer was collected in a 500 mL Erlenmeyer flask. The mixture was extracted three times with 25 mL of acetic acid, and then the lower layer was collected in a 500 mL Erlenmeyer flask. 25 mL of acetic acid was added thereto, 1 mL of saturated KI solution was added thereto, followed by shaking for 1 minute. The mixture was left for 10 minutes in the dark, 50 mL of distilled water was added thereto, and titrated with 0.01 N sodium thiosulfate solution using 1% starch solution as an indicator. It was.

S: 0.01 N-Na ₂ S ₂ O ₃ titration (mL)

N: prescribed concentration of Na ₂ S ₂ O

L: weight of fat or oil (g)

As a result, as shown in FIG. 1, NaCl increased with increasing concentration, and the POV increased significantly at 0.6%, but KCl was hardly increased to the same value as the control.

Experimental Example 2: Comparison of Mutagenicity between NaCl and KCl

In this experimental example, NaCl and KCl were treated with Salmonella typhimurium TA100 strain with MNNG at concentrations of 3, 7, 15, and 25% by Ames test to compare the mutagenicity of NaCl and KCl. The Ames test has a positive correlation (85%) between animal carcinogenesis experiments in vivo . The preincubation test mainly used in this experiment was 0.5 mL of S9 mix (if indirect mutation) or 0.5 mL of phosphate buffer (if direct mutation), Salmonella typhimurium TA100 strain (1-2 × 10 ⁹ cells) cultured overnight. / ML) 0.1mL, diluted sample NaCl, KCl (50µL) and mutagen (50µL) were put in a cap tube in an ice bath, lightly vortexed, and then 37 ° C. Pre-incubation for 30 minutes at. Pour 2 mL of top agar at 45 ° C into each tube, vortex for 3 seconds, spread on minimal glucose agar plate, incubate at 37 ° C for 48 hours, and return mutation number. Was counted. Meanwhile, the concentrations of samples and mutagenic substances used in the experiments were determined through preliminary experiments (dose response and toxicity experiments). The inhibition rate (mutation rate) was calculated by the following equation.

% Inhibition = 100 × [(a­b) / (a­c)]

Where a is the number of back mutations induced by the mutagen, b is the number of back mutations when the sample is processed, and c is the number of back mutations when no mutagen and the sample are present.

As a result, 3% treatment resulted in 695 ± 27 return mutations but NaCl increased 1445 ± 136, but KCl increased 798 ± 35 and 7% treatment did not significantly increase return mutations. 204 were found, but KCl was 892 +/- 86, and the mutational variability was significantly different. The same tendency was observed in 15 and 25%. This suggests that Na is mainly involved in inducing mutagenic mutations, but Cl is also involved in some degree.

Example 2: Inhibition of Cytotoxicity and MCC on MCA in C3H10T1 / 2 Cells of NaCl and KCl

The culture of the cells and cells used in the present invention is C3H / 10T1 / 2 mouse mouse cells (Mouse embryo cells) containing 100 unit / mL penicillin-streptomycin (penicillin-streptomycin) and 10% FCS (fetal calf serum) Cultured in a 37%, 5% CO ₂ incubator using basal medium Eagle (BME). Cultured C3H / 10T1 / 2 cells are refeeded every 5 days, detached cells attached with 0.05% trypsin-0.02% EDTA enzyme with PBS solution, and subdivided in a cell culture fask. It was used for the experiment while incubating. The dose response was inoculated with C3H / 10T1 / 2 cells in a 60mm dish at a concentration of 2,000 cells / 5mL, incubated for 24 hours, discarded the culture medium, and the MCA (1, 5, 10) in free serum medium. , 50 ㎍ / mL) was added to each 60mm dish after 48 hours. The experimental and control (DMSO) groups were reloaded with fresh medium containing 10% FCS. After one week of incubation, the cells were immobilized with methanol, stained with Giemsa stain, and the colonized cell colonies were counted. Cytotoxicity test was inoculated with C3H / 10T1 / 2 cells in a 60mm dish at a concentration of 2,000 cells / 5mL, incubated for 24 hours, discarded the culture medium, and carcinogenic to free serum medium. Substances [MCA (5µg / mL)], NaCl and KCl samples were added, and 5mL each was inoculated into a 60mm dish. The experimental and control groups were refeeded with fresh medium containing 10% FCS. After one week of incubation, the cells were immobilized with methanol, stained with Giemsa stain, and counted in colony cell colonies to measure the cytotoxicity effects of NaCl and KCl on C3H / 10T1 / 2 cells according to the following formula. It was.

In addition, C3H / 10T1 / 2 cells were counted at 2,000 cells / 5mL and seeded in a 60mm dish, and cultured for 24 hours to examine the effect of NaCl and KCl on MCA transformation. Discard the culture medium and put the MCA (20-methylcholanthrene) and the sample in a free serum medium and incubated for 24 hours, carcinogen and H ₂ O was added to the control. Treatment and control groups were incubated in 37 ° C., 5% CO ₂ incubator for 5 weeks with refeeding with fresh medium containing 10% FCS. Thereafter, the cells were immobilized with methanol and stained with Giemsa stain, and counted according to three types (type I, II, III) according to the intensity of transformation of the transformed foci. As shown in Table 1, NaCl and KCl inhibited the cytotoxicity of MCA C3H / 10T1 / 2 cells at all concentrations. KCl showed higher cytotoxicity inhibitory effect than NaCl, and the addition of 30% NaCl dramatically reduced cytotoxicity, suggesting that high concentrations of salt (NaCl) may have a cocarcinogenic effect. . In order to examine the effects of MCA on the formation / transduction of transformed foci, the results of counting foci classified into Type I, Type II, and Type III were as shown in Table 2. In the control group, 35 were Type II (50% carcinogenic) and Type III (85% carcinogenic), whereas KCl was 20 and 26 at 20% and 30% concentrations, respectively. It was observed that KCl exhibited cancer suppression effect by inhibiting formation. In particular, KCl had a great effect on the inhibition of Type III posiogenesis. However, NaCl decreased the formation of type I poems, but increased the formation of type II + type III poems (40), suggesting that high concentrations of salt may act as a cancer-causing agent. Therefore, KCl substitution Kimchi used in this experiment was considered to be advantageous in terms of cancer prevention.

Cytotoxic Inhibitory Effect of NaCl and KCl on MCA in C3H / 10T1 / 2 Cells Treatment group (%) Cell colony count One 5 10 20 30 MCA (control) 108 ± 4 (1.00) ¹⁾ NaCl 115 ± 3 (1.06) 124 ± 3 (1.15) 139 ± 2 (1.29) 144 ± 4 (1.33) 119 ± 2 (1.10) KCl 130 ± 3 (1.20) 151 ± 4 (1.40) 159 ± 6 (1.47) 163 ± 3 (1.51) 165 ± 1 (1.53) ¹⁾ Values in parentheses indicate cytotoxicity

Inhibitory Effects of NaCl and KCl on MCA in C3H / 10T1 / 2 Cells Sample(%) Posai produce number Type I Posay Type II Posay TypeⅢ Posay Type II + III Posay MCA (Control) 28 ± 3 25 ± 2 10 ± 2 35 ± 2 MCA + NaCl 20 23 ± 3 25 ± 3 15 ± 3 40 ± 3 30 23 ± 2 20 ± 2 16 ± 2 36 ± 3 MCA + KCl 20 35 ± 2 16 ± 2 4 ± 4 20 ± 4 30 31 ± 3 17 ± 4 8 ± 2 26 ± 4

Example 3 KCl Kimchi Preparation of the Invention

In this example, to reduce the salt content, which is the most problematic problem in kimchi, 30% of the amount of salt added was prepared with KCl. As shown in Table 3, kimchi with 30% of salt added to the standardized Chinese cabbage kimchi and cancer prevention cabbage kimchi recipe was replaced with KCl, fermented at 15 ℃ for 1 day, and then fermented at 5 ℃ for physicochemical changes. After the lyophilized kimchi was lyophilized and extracted with methanol, the effects of cancer prevention and anticancer activity were examined. First, the Chinese cabbage was divided into 4 equal parts and washed three times with salt water (10%) twice the weight of Chinese cabbage at 5 ° C and washed three times, followed by draining for 2 hours. In case of salting with different kinds of salt, salt water was made with sun salt, Hangeumgeum, and roasted salt, and each was adjusted to 10%. Standardized Chinese cabbage kimchi used general cabbage (Bulam No. 3) and nutritive solar herb, and cancer prevention functional enhancement kimchi was made of organic cabbage (Bulam No. 3) and organic method of red pepper powder. The kimchi mixed with the recipe was put into 1 kg bottles in 1 kg, fermented at 15 ° C. for 1 day, and then fermented at 5 ° C. for 6 weeks. 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 4, 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, the inhibitory effect of DNA synthesis using HT-29 cancer cells was shown in Table 5 at concentrations of 10 μl / well, NaCl 28%, mechanical salt 40%, sun salt 45%, fresh salt 30%, baked salt 82%, and salt Inhibition of DNA synthesis by 83% and bamboo salt by 61% was also observed.

Components of Various Cabbage Kimchis Ingredient (g) Standardized Kimchi (SK) Functional Kimchi (FK) SK Ⅰ SK II SK Ⅲ SK Ⅳ FK II FK I Sun salt Natural salt + KCl Baked Salt + KCl One address + KCl Baked Salt + KCl One address + KCl cabbage 100.0 100.0 100.0 100.0 100.0 100.0 chili powder 3.5 3.5 3.5 3.5 7.0 7.0 chopped garlic 1.4 1.4 1.4 1.4 2.8 2.8 Chopped ginger 0.6 0.6 0.6 0.6 0.6 0.6 Anchovy Sauce 2.2 2.2 2.2 2.2 2.2 2.2 Sugar 1.0 1.0 1.0 1.0 1.0 1.0 radish 13.0 13.0 13.0 13.0 13.0 13.0 wave 2.0 2.0 2.0 2.0 2.0 2.0 lampshade 5.0 5.0 Chopi 0.1 0.1 Sun salt 1.0 Baked Salt 1.0 KCl 1.0 1.0 1.0 1.0 * Uses natural salt for pickling * Uses natural salt for pickling * Baked salt is used for pickling * One address is used for pickling * Baked salt is used for pickling * Baked salt is used for pickling

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 4 Antimutagenicity of KCl Chinese Cabbage Kimchi of the Present Invention in vitro Anticancer effect

Experimental Example 1: Physicochemical and Sensory Properties

The fermentation of cabbage kimchi prepared in Example 3 was carried out to investigate the pH and acidity, Leuconostoc sp. Lactobacillus sp. Involved in lactic acid bacteria and acid production. As a result of measuring the amount of lactic acid bacteria in a week, standardized kimchi II (1.7% natural salt + 0.8% KCl) and standardized kimchi III (1.7% roasted salt + 0.8) were substituted for KCl in the standard cabbage kimchi recipe as shown in FIG. % KCl) and Standardized Kimchi IV (1.7% Hanjugeum + 0.8% KCl) were slower to fermentation than Standardized Kimchi I (2.5% Natural Salt), while Standardized Kimchi I (2.5% Natural Salt) had a pH of 3.99 at 4 weeks. Kimchi II, III, and IV had a pH above 4.0 even at 5 weeks. Kimchi I and II promoted cancer prevention function and fermentation was slower than standardized kimchi with pH of 4.23 and 4.38, respectively. Also, as shown in FIG. 3, the standardized kimchis II, III, and IV replacing KCl were lower than the standardized kimchi I (2.5% sun salt), and the acidity of the cancer preventive function enhancing kimchi II was the lowest during the whole fermentation. Leuconostoc sp. Involved in taste. As shown in FIG. 4, lactic acid bacteria had more standardized kimchi II, III, and IV than standardized kimchi I (2.5% sun salt) at 1 week of fermentation, but there was no significant difference thereafter. Standardized Kimchi I, II, III, and IV showed Leuconostoc sp. While increasing the number of lactic acid bacteria and then decreased cancer promoting functional kimchi Ⅰ, Ⅱ are Leuconostoc than kimchi were decreased after 5 weeks dipped to promote cancer prevention, functional kimchi recipe dipped into standardized recipes sp. The number of lactic acid bacteria was rather low. As shown in Fig. 5, the standardized kimchis II, III, and IV had less Lactobacillus sp. Lactic acid bacteria than standardized kimchi I during fermentation. Therefore, as shown in Fig. 6, the standardized kimchis II, III, and IV, which are KCl alternative kimchi, had a bitter bitter taste, and thus, the overall evaluation was not good.

Experimental Example 2: Antimutagenic Enhancing Effect and Cytotoxic Inhibitory Effect on MCA in C3H10T1 / 2 Cells

In this Experimental Example, except that the sample using the cabbage kimchi methanol extract prepared in Example 3, and the antimutagenic effect and the cytotoxic effect on MCA in C3H10T1 / 2 cells by the same method as in Example 2 Was investigated. In other words, the antimutagenic effect of AFB1 of Salmonella typhimurium TA100 strains of 6 kinds of Chinese cabbage kimchi methanol extracts soaked with KCl in the standard recipe and the cancer prevention function enhancement recipe was investigated. As a result, 1.25mg / Standardized kimchi I (2.5% natural salt) at plate addition concentration inhibited the mutation of AFB1 by 53%, while standardized kimchi II (1.7% natural salt + 0.8% KCl) and standardized kimchi III (1.7%) replaced KCl in standard recipe. Baked salt + 0.8% KCl) and standardized Kimchi IV (1.7% Hanjugeum + 0.8% KCl) showed 69% antimutagenic effect and cancer preventive function enhancing kimchi II showed 74% antimutagenic effect. More than 20% of the antimutagenic effect of 2.5% natural salts). However, there was no difference in antimutagenic activity according to the type of salt used for pickling. In addition, Table 7 shows the results of cytotoxicity test showing that KCl replacement kimchi inhibits cytotoxicity by MCA in C3H / 10T1 / 2 cells. Cytotoxicity can be determined by the degree of inhibition of colony formation. The standardized kimchi I (2.5% sun salt) showed 23% cytotoxicity at standard concentration of / mL and standardized kimchi II (1.7% sun salt + 0.8% KCl) and standardized kimchi III (1.7% baked salt + 0.8% KCl). And Kimchi IV (1.7% Hanjugeum + 0.8% KCl) showed a cytotoxic effect of 41-44%. Kimchi substituted KCl had a greater cytotoxic effect by MCA. The high cytotoxicity inhibitory effect of% showed that cancer prevention function could be further enhanced by substituting KCl for cancer prevention function enhancement recipe.

Antimutagenic Effects of KCl Chinese Cabbage Kimchi Methanol Extract on AFB ₁ of Salmonella typhimurium TA100 Strain Treatment (mg / plate) Return mutation / plate 0.625 1.25 Spontaneous 89 ± 9 AFB ₁ (Control) 1043 ± 34 ^a Standardized Chinese Cabbage Kimchi I (Sun Salt) 687 ± 23 ^b (37) ¹⁾ 540 ± 33 ^b (53) Standardized Chinese Cabbage Kimchi Ⅱ (Sun Salt + KCl) 545 ± 39 ^d (52) 388 ± 11 ^d (69) Standardized Chinese cabbage kimchi Ⅲ (grilled salt + KCl) 506 ± 24 ^d (56) 380 ± 14 ^d (69) Standardized Chinese cabbage kimchi IV (Hanjugeum + KCl) 620 ± 24 ^c (44) 386 ± 6 ^d (69) Cancer Prevention Functional Kimchi ²⁾ Ⅰ (Baked Salt) 512 ± 13 ^d (56) 438 ± 23 ^c (63) Cancer Prevention Functional Kimchi Ⅱ (Grilled Salt + KCl) 421 ± 14 ^e (65) 330 ± 10 ^e (74) ¹⁾ The values in parentheses indicate significant differences between the inhibition rates ^{a ~ d} in Duncan's multiple range test.

KCl Alternative Kimchi Inhibits MCA-induced Cytotoxicity in C3H / 10T1 / 2 Cells Treatment group (50 µg / mL) Cell colonies Cytotoxicity MCA (Control) 98 ± 7 ^a 1.00 Standardized Chinese Cabbage Kimchi I (Sun Salt) 121 ± 3 ^b 1.23 Standardized Chinese Cabbage Kimchi Ⅱ (Sun Salt + KCl) 139 ± 2 ^c 1.41 Standardized Chinese cabbage kimchi Ⅲ (grilled salt + KCl) 142 ± 2 ^c 1.44 Standardized Chinese cabbage kimchi IV (Hanjugeum + KCl) 139 ± 2 ^c 1.41 Cancer prevention functional kimchi Ⅰ (grilled salt) 141 ± 3 ^c 1.44 Cancer Prevention Functional Kimchi Ⅱ (Grilled Salt + KCl) 156 ± 2 ^d 1.59 ^{a to d} represent significant differences in Duncan's multiple range test.

Experimental Example 3: In vitro Anticancer activity

In this experimental example, in vitro anticancer activity of AGS human gastric cancer cells of six kinds of Chinese cabbage kimchi methanol extracts substituting KCl for standard cabbage kimchi recipe and cancer prevention functional cabbage kimchi recipe in Example 3 was examined.

AGS cancer cells were inoculated in a 96 well plate at 1 × 10 ³ cells / mL per well, and cultured for 24 hours, followed by the addition of the sample extract, followed by incubation in a 37 ° C. 5% CO ₂ incubator. After 3 days of culture, 50 μl of MTT was added and further incubated for 4 hours, and then 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 reduced cell growth by 50% was calculated according to the Lotus data regression program. In addition, cancer cell proliferation inhibition experiment was carried out in the same manner as in the culture of cancer cells, but AGS cancer cells, which are adherent cells, were centrifuged and mixed well so that the accumulated cancer cells were evenly distributed. After confirming that the cancer cells were attached to the plate, the sample was added to the medium containing 10% FCS and cultured in a 37 ° C., 5% CO ₂ incubator while replacing the culture solution every other day. Samples were replaced with fresh medium added at 10 μl / mL medium every 2 days. After 6 days of incubation, the cells were grown with 0.05% trypsin-0.02% EDTA enzyme and the number of cells was hemocytometer. The cancer cell proliferation inhibitory effect was observed in comparison with the control group. As shown in Table 8, the standardized kimchi Ⅰ (2.5% sun salt) showed 45% gastric cancer cell proliferation inhibitory effect at 100 ㎍ / assay, while standardized kimchi Ⅱ (1.7% sun salt) replaced KCl in the standard cabbage kimchi recipe. + 0.8% KCl), Standardized Kimchi III (1.7% Baked Salt + 0.8% KCl) and Standardized Kimchi IV (1.7% Hanbok + 0.8% KCl), 54 ~ 58%, Kimchi II improved cancer prevention 75% Proliferation inhibitory effect was shown. Table 9 shows the effect of inhibiting the growth of AGS human gastric cancer cells after 6 days of treatment with KCl cabbage kimchi methanol extract sample, 30% higher than standardized kimchi I (2.5% natural salt). It showed higher growth inhibitory effect on gastric cancer cells.

Inhibitory Effect of KCl Chinese Cabbage Kimchi Methanol Extract on AGS Human Gastric Cancer Cell by MTT Method Treatment group (㎍ / assay) OD ₅₄₀ 100 200 Control 0.621 ± 0.020 ^a 0.621 ± 0.020 ^a Standardized Chinese Cabbage Kimchi I (Sun Salt) 0.339 ± 0.010 ^b (45) ¹⁾ 0.115 ± 0.010 ^b (82) Standardized Chinese Cabbage Kimchi Ⅱ (Sun Salt + KCl) 0.283 ± 0.010 ^c (54) 0.055 ± 0.010 ^c (91) Standardized Chinese cabbage kimchi Ⅲ (grilled salt + KCl) 0.263 ± 0.010 ^cd (58) 0.039 ± 0.010 ^c (94) Standardized Chinese cabbage kimchi IV (Hanjugeum + KCl) 0.281 ± 0.010 ^cd (55) 0.049 ± 0.010 ^c (92) Cancer prevention functional kimchi Ⅰ (grilled salt) 0.259 ± 0.010 ^d (58) 0.049 ± 0.010 ^c (92) Cancer Prevention Functional Kimchi Ⅱ (Grilled Salt + KCl) 0.158 ± 0.010 ^e (75) 0.012 ± 0.010 ^d (98) One) Subscripts ^{a to e} show significant differences by Duncan's multiple range test.

Growth Inhibitory Effect of KCl Chinese Cabbage Kimchi Methanol Extract on AGS Human Gastric Cancer Cells Treatment group (50 µg / mL) Cell number (× 10 ⁴ / mL) Inhibition Ratio (%) Control 70 ± 4 ^a Standardized Chinese Cabbage Kimchi I (Sun Salt) 35 ± 2 ^b 50 Standardized Chinese Cabbage Kimchi Ⅱ (Sun Salt + KCl) 20 ± 3 ^c 71 Standardized Chinese cabbage kimchi Ⅲ (grilled salt + KCl) 19 ± 1 ^c 73 Standardized Chinese cabbage kimchi IV (Hanjugeum + KCl) 22 ± 2 ^c 69 Cancer prevention functional kimchi Ⅰ (grilled salt) 19 ± 2 ^c 73 Cancer Prevention Functional Kimchi Ⅱ (Grilled Salt + KCl) 10 ± 2 ^d 86 Subscripts ^{a to e} show significant differences by Duncan's multiple range test.

Example 5: In vivo Antitumor and Immune Activity of KCl-Containing Chinese Cabbage Kimchi in Korea

In this example, in order to examine the effects of NaCl and KCl in the in vivo chemotherapy system, using mouse sarcoma-180 tumor cells, the effect of inhibiting solid cancer growth in the mouse, organ weight changes such as liver and spleen, changes in spleen immune function, liver The changes in glutathione-S-transferase activity present were examined. The experimental animals used in this Example were male Balb / c mice weighing about 25 g for anticancer experiments, and the feed was raised as standard feed. During breeding, water and feed were supplied in sufficient quantities, and the animal room maintained a light-dark cycle of 12 hours.

The tumor cells used in this example were taken with ascites sarcoma-180 cells cultured for one week in the abdominal cavity of the experimental animal and centrifuged (1,200rpm, 10min) with phosphate buffered saline (PBS). Was separated. The separated cells were suspended in PBS again, centrifuged again to remove the supernatant, and tumor cells were suspended in sarcoma-180 cells to 1.0 × 10 ⁶ cells / ml.

Samples used in this example were prepared using sterile PBS, the dosage was 0.5 mg per mouse, and the control group was injected with sterile PBS only and stored in the refrigerator when not administered.

Experimental Example 1: Cytotoxicity Test

In order to investigate the direct cytotoxicity of kimchi samples, a viability test was performed in vitro using the dye exclusion method. 24 well plates were inoculated 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. and 5% CO ₂ 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 non-stained cells using a hemocytometer. After counting, the survival rate was calculated by comparison with the control cell group without the kimchi sample.

As a result, as shown in Table 10, all the samples were not toxic to 1.0 mg / mL as a result of the viability test, so that 1.0 mg / kg of the sample was administered to the abdominal cavity of the mouse to examine the effect of inhibiting solid cancer growth. In addition, as shown in Table 11, the weight of each organ was similar to that of the control group transplanted with tumor cells, but the standardized kimchi Ⅱ (1.7% natural salt + 0.8% KCl) and the standardized kimchi were shown. The group administered with III (1.7% roasted salt + 0.8% KCl) gained more weight than the tumor cell transplant group. In the control group transplanted with tumor cells only, the weight ratio of spleen, liver, and kidney was significantly higher than that of normal mice, and the standardized kimchi Ⅰ (2.5% sun salt) group showed similar weight ratio to tumor cell transplant group, but the standardized kimchi Ⅱ (1.7% natural salt + The weight ratio of liver, kidney, and spleen was decreased in the group receiving 0.8% KCl) and standardized Kimchi III (1.7% roasted salt + 0.8% KCl).

Viability of Sarcoma-180 Cells in Medium Containing Various Kimchi Methanol Extracts Sample Final concentration (mg / mL) Total cell number (× 10 ⁴ ) Viable cells (%) Control 23.7 ± 1.2 98 Standardized Kimchi Ⅰ 0.1 23.3 ± 0.9 97 (Sun salt) 0.5 22.0 ± 0.8 92 1.0 20.0 ± 0.8 83 1.5 19.3 ± 0.9 79 Standardized Kimchi Ⅱ 0.1 24.0 ± 0.8 96 (Natural salt + KCl) 0.5 23.0 ± 0.8 94 1.0 21.0 ± 0.8 86 1.5 19.7 ± 0.9 80 Standardized Kimchi Ⅲ 0.1 23.7 ± 0.5 98 (Baked Salt + KCl) 0.5 22.0 ± 0.8 92 1.0 21.7 ± 1.2 89 1.5 18.3 ± 0.5 76 Functional Kimchi 0.1 23.7 ± 0.5 98 (Baked Salt + KCl) 0.5 21.7 ± 0.5 89 1.0 21.0 ± 0.8 86 1.5 18.3 ± 0.9 73

Weight Changes of Organs in Bal / c Mice Treated with Various Kimchi Methanol Extracts Sample Dose (mg / kg) Weight (g) Spleen / Weight (%) Liver / weight (%) Heart / weight (%) Height / weight (%) Control 30.1 ± 0.9 0.3 ± 0.1 6.3 ± 0.6 0.5 ± 0.0 1.5 ± 0.1 S-180 + PBS 28.6 ± 0.7 1.5 ± 0.1 8.4 ± 0.5 0.3 ± 0.0 1.9 ± 0.1 S-180 + SK ¹ Ⅰ (Sun Salt) 1.0 28.0 ± 1.2 1.5 ± 0.1 7.9 ± 0.3 0.5 ± 0.1 1.9 ± 0.1 S-180 + SK ¹ Ⅱ (Sun Salt + KCl) 1.0 29.6 ± 0.8 1.2 ± 0.1 7.7 ± 0.5 0.5 ± 0.0 1.7 ± 0.1 S-180 + SK ¹ Ⅲ (Baked Salt + KCl) 1.0 29.5 ± 0.1 1.2 ± 0.1 7.1 ± 0.5 0.5 ± 0.1 1.6 ± 0.1 S-180 + FK ² (Baked Salt + KCl) 1.0 28.9 ± 0.5 1.2 ± 0.1 6.6 ± 0.2 0.5 ± 0.0 1.7 ± 0.1

Experimental Example 2: Effect of Blocking Solid Cancer Growth

10 animals per group were injected into the left groin of 0.2 mL (6 × 10 ⁶ cells / mouse) of tumor cell suspension stored in the laboratory for subcutaneous transplantation. Sample solutions were administered intraperitoneally. On the 32nd day of tumor cell transplantation, the resulting solid cancer was extracted and weighed. As a result, as shown in Table 12, the control group transplanted with only tumor cells had a tumor weight of 6.1 g, but the standardized Kimchi I (2.5% sun salt) -administered group had a tumor weight suppression effect of 4.7% due to a decrease in tumor weight of 4.7 g. Indicated. Standardized Kimchi Ⅱ (1.7% natural salt + 0.8% KCl) Kimchi group showed 39% higher tumor suppression effect than standardized Kimchi I. When Kimchi was replaced with salt (NaCl), the effect of suppressing solid cancer growth Elevation could be observed. The functional kimchi group had a tumor weight of 3.0 g, showing 52% tumor suppression effect, and showed 2 times more solid cancer growth inhibition than the standard Kimchi I group.

Inhibitory Effects of Kimchi Methanol Extract on Solid Cancer Cell Growth in Balb / c Mice Implanted with sarcoma-180 Cells Sample Dose (mg / kg) Tumor wt. (G) Inhibition Ratio (%) S-180 + PBS 6.1 ± 0.1 S-180 + SKⅠ (Sun Salt) 1.0 4.7 ± 0.3 23 S-180 + SKII (Sun Salt + KCl) 1.0 3.8 ± 0.3 39 S-180 + SKⅢ (Baked Salt + KCl) 1.0 4.1 ± 0.1 33 S-180 + FK (Baked Salt + KCl) 1.0 3.0 ± 0.3 52

Experimental Example 3: Glutathione-S-transferase (GST) activity change

After the mice were killed, the liver was perfused with saline of 4 ° C. or less 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 ground with a glass teflon homogenizer under ice cooling. The homogenate fraction was used as a homogenate fraction, which was centrifuged at 13,000 rpm for 10 minutes to remove the nucleus and unbroken cell sections, and the supernatant obtained by ultracentrifugation at 105,000 × g for 1 hour was subjected to cytosol ( cytosol) fraction. 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, 75 μl of reduced glutathione with 0.04 M reduced glutathione was added to 0.1 M potassium phosphate buffer (pH 6.5), and 0.1 mL of the enzyme solution was added to the blank. 20% trichloroacetic acid was added to the blank. acid) Inactivate the enzyme by adding 0.5 mL, and react the sample at 25 ℃ for 5 minutes, add 25 μl of 0.12M 1-chloro-2,4- dinitrobenzene to the blank and sample, and react at 25 ℃ for 2 minutes. 20% trichloroacetic acid was added to complete the reaction, and the supernatant obtained by centrifugation was measured for absorbance at 340 nm, followed by 1-chloro-2,4-dinitrobenne (1-chloro-2,4-dinitroben). The activity was calculated using the mole extinction coefficient of zene) of 9.6 mM ^-1 cm ^-1 . The unit of enzyme activity was expressed as nmole number of 2,4-dinitrobenzene-glutathione produced by mg protein for 1 minute. As shown in FIG. 6, the GST activity was 613.5 nmol / mg protein / min, the highest in the normal group without transplanting tumor cells, and the control group in which tumor cells were transplanted was reduced to 290.0 nmol / mg protein / min. The standardized kimchi I (2.5% natural salt) group was 406.3 nmol / mg protein / min, which showed higher GST activity than the tumor cell transplant group, and standardized kimchi II (1.7% natural salt + 0.8% KCl) and standardized kimchi III (1.7% roasted salt + 0.8% KCl) was 447.9 nmol / mg protein / min and 451.4 nmol / mg protein / min, respectively, higher than the standardized Kimchi I (2.5% natural salt) group. The GST activity of the functional kimchi-administered group was 477.9 nmol / mg protein / min, which was higher than the standardized kimchi II (1.7% natural salt + 0.8% KCl) and standardized kimchi III (1.7% baked salt + 0.8% KCl).

Experimental Example 4: Immune Activity Enhancing Effect

To isolate mouse spleen lymphocytes, spleens of Balb / c mice were aseptically extracted on a clean bench, containing penicillin (100 U / mL) and streptomycin (100 μg / mL). Three washes with 5 mL of RPMI 1640 medium followed by fine grinding. The cell suspension was filtered through a 70 μm nylon mesh, centrifuged to collect lymphocytes, and then suspended in the same medium. Lytopocytes were separated by centrifugation (500 × g, 30 min, 18 ° C.) using histopaque-1077. Effector cells were resuspended in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamin, penicillin (100 U / ml), streptomycine (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-adherent natural killer (NK) cells were collected by centrifugation, resuspended in culture medium, and the appropriate cell number was counted. Activity analysis of natural killer cells was carried out using the MTT method of Experimental Example 3 of Example 4. Here, 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-glutamin, penicillin (100 U / ml), and streptomycine (100 µg / ml). 50 μl was added to each well. NK cells were suspended at a density of 1 × 10 ⁷ cells / ml. For NK cell activity analysis, 50 μl of effector cells and Yac-1 cells were added to a 96well flate-bottomed microtire plate. This was incubated for 3 days at 37 ° C. in a CO ₂ incubator, and 10 μl of MTT (5 mg / ml) solution was added to each well, followed by incubation 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 cell lysis was calculated by the following formula.

As shown in FIG. 8, the cell destruction activity of Yac-1 cells was 41% in the standardized Kimchi I (2.5% sun salt) -treated group, and standardized Kimchi II (1.7% sun salt + 0.8% KCl) and standardized Kimchi III (1.7). In the group treated with% grilled salt + 0.8% KCl), the KCl alternative kimchi group was higher in NK cells than the standard Kimchi I (2.5% natural salt) group. Also, the functional kimchi-treated group showed high cell destruction activity of 55%. Substituting KCl for cancer preventive functional promotion recipe, cell destruction activity against natural killer cells in mouse spleen was increased compared to standardized Kimchi I (2.5% natural salt). Confirmed high.

Example 6: Inhibitory effect on tumor metastasis of KCl-containing cabbage kimchi of the present invention

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. In order to measure the tumor metastasis inhibition effect of KCl cabbage kimchi, cultured colon 26-M3.1 carcinoma (handong university, Dr. Yoon Taek-jun, Ph.D.), obtained metastasis to the lung, in virto (7.5% FBS contained) 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 shown in Table 13, the standardized kimchi I (2.5% sun salt) group received 14% subcutaneous injection of 1.25 mg / mouse in the experimental result of the tumor metastasis using Colon 26-M-3.1 carcinoma. Showed low tumor metastasis inhibitory effect. Promoting cancer prevention function Kimchi I showed a 12% metastasis inhibitory effect similar to that of the standardized Kimchi I group even at a low concentration of 0.05 mg. Growth of colon 26-M3.1 tumors that metastasized to the lung when subcutaneous injection of 1.25 mg / mouse Was significantly inhibited to about 35%, and it was confirmed that kimchi also has a preventive effect of cancer metastasis. In addition, cancer preventive functional kimchi Ⅱ (Gueun salt + KCl) showed significant tumor metastasis inhibitory effect at all concentrations of 0.05 ~ 1.25mg / mouse and 49% high metastasis inhibitory effect when 1.25mg / mouse was administered. It was confirmed that the substitution of KCl in the cancer prevention functional enhancement recipe can increase the preventive effect of cancer metastasis. Figure 9 showed a significant difference as shown in the tumor metastasis inhibitory effect of the control and cancer prevention functional enhanced kimchi II (Gueun salt + KCl) treated with methanol extract of colon 26-M3.1 cells.

Tumor Metastasis Inhibitory Effect of Highly Active Chinese Cabbage Kimchi Using Colon 26-M3.1 Cells Treatment Dose (mg / mouse) root Lung carcinogens (% inhibition) Mean ± SD range Control sc 162 ± 7 ^a 153 ~ 172 Standardized Kimchi I (Sun Salt) 0.050.251.25 scscsc 157 ± 13 ^a (3) ¹⁾ 147 ± 8 ^ab (9) 139 ± 5 ^bc (14) 142 ~ 172138 ~ 157131 ~ 144 Cancer prevention functional kimchi I (grilled salt) 0.050.251.25 scscsc 143 ± 12 ^ab (12) 125 ± 15 ^ab (23) 103 ± 5 ^e (36) 124 ~ 156109 ~ 14696 ~ 108 Cancer Prevention Functional Kimchi Ⅱ (Grilled Salt + KCl) 0.050.251.25 scscsc 119 ± 4 ^d (27) 99 ± 8 ^e (39) 83 ± 6 ^t (49) 114 ~ 12389 ~ 11073 ~ 91 1) Subscripts show significant differences by Duncan's multiple range test.

As described above, the salt and cabbage 100, pickled cabbage 100%, leek 2.0%, red pepper powder 3.5%, garlic 1.4%, ginger 0.6%, anchovy 2.2%, sugar 1.0%, final salt 2.5 For standardized cabbage kimchi or pickled cabbage containing 100%, pepper powder 7%, garlic 2.8%, chopi 0.1%, fresh 5%, ginger 0.6%, radish 13.0%, green onion 2.0%, anchovy sake 2.2%, sugar 1.0%, Enhancement of cancer prevention function containing 2.5% of the final salt By replacing 20-40% of the salt added to kimchi with KCl, it has an excellent effect of improving anticancer properties without changing the physicochemical and sensory properties of Chinese cabbage kimchi. It is a very useful invention.

Claims (6)

Salt added in standardized cabbage kimchi containing 13.0% radish, 2.0% leek, 3.5% red pepper powder, garlic 1.4%, ginger 0.6%, salted anchovy 2.2%, sugar 1.0% and final salt 2.5% KCl-containing anti-cancer cabbage kimchi, characterized in that 30% of the replacement with KCl. [Claim 2] The anti-cancer cabbage kimchi containing KCl according to claim 1, wherein the salt selects any one or more selected from processed salts consisting of roasted salt, fresh salt, and bamboo salt. In preparing standardized Chinese cabbage kimchi containing 100% radish 13.0%, leek 2.0%, red pepper powder 3.5%, garlic 1.4%, ginger 0.6%, anchovy salt 2.2%, sugar 1.0%, final salt 2.5% It is fermented at 15 ℃ for 1 day, and then fermented at 5 ℃ for 6 weeks in a pickled cabbage with red pepper powder, garlic, chopi, fresh, sugar, green onion, radish mixed with KCl so that 30% of the total salt content. KCl-containing anti-cancer cabbage kimchi production method characterized in that. Cancer containing 100% pickled cabbage, red pepper powder 7%, garlic 2.8%, chopi 0.1%, fresh 5%, ginger 0.6%, radish 13.0%, green onion 2.0%, anchovy salt 2.2%, sugar 1.0%, final salt 2.5% Preventive Functional Enhancement In kimchi, anti-cancer cabbage kimchi containing KCl, characterized in that 30% of added salt is replaced with KCl. 5. The KCl-containing anti-cancer cabbage kimchi according to claim 4, wherein the salt selects any one or more from processed salts consisting of roasted salt, fresh salt, and bamboo salt. Cancer containing 100% pickled cabbage, red pepper powder 7%, garlic 2.8%, chopi 0.1%, fresh 5%, ginger 0.6%, radish 13.0%, green onion 2.0%, anchovy salt 2.2%, sugar 1.0%, final salt 2.5% Preventive Functional Enhancement In making kimchi, fermented at 15 ° C for 1 day with salt and cabbage, pickled cabbage, garlic, chopi, fresh, sugar, green onion and radish mixed with KCl 30% of the total salt. KCl-containing anti-cancer cabbage kimchi manufacturing method characterized in that the fermentation for 5 weeks at 5 ℃.