KR20010044190A - Improvement effect of diets supplemented with Chinese cabbage having a high concentration of GABA on lipid metabolism and liver function of rats administered with ethanol chronically - Google Patents

Abstract

PURPOSE: A cabbage-contained diet containing high percentage of GABA(gamma-Aminobutyric Acid) is provided, which shows improvement effect on fat metabolism and liver function of a chronic alcohol-administrated white mouse. CONSTITUTION: The fat metabolism and liver function improvement effect of cabbage-contained diet containing high percentage of GABA on chronic alcohol-administrated white mouse is observed by the following processes of: collecting summer cabbage, washing and freeze-drying; pulverizing cabbage leaves and cabbage roots into powder using liquid nitrogen gas, and adding a mixed solution of methanol:chloroform:water(12:5:3) to measure content of GABA and amino acid; centrifuging to get the solution layer containing free amino acid and GABA, adding a mixed solution of chloroform:water(5:3) to the residue to extract remaining free amino acid and GABA; gathering the solutions and freeze-drying, and dissolving with small quantity of water; filtering with 0.45μm PVDF filter, analyzing with an Amino Acid Analysis System, and comparing with standard amino acid and standard GABA to calculate content of GABA and amino acid.

Description

Improvement effect of diets supplemented with Chinese cabbage having a high concentration of GABA on lipid metabolism and liver function of rats administered with ethanol chronically}

Gamma-aminobutyric acid (GABA) is gaining attention as a source of new functional food as well as as a dietary supplement, and it can be used as a food material by accumulating a large amount of gamma-aminobutyric acid during germination of rice embryos. A method for producing gamma-aminobutyric acid is known, and a skim food material in which gamma-aminobutyric acid is accumulated in a large amount during the germination process is used. The general development process related to GABA is as follows.

During the study of the biologically active components of rice embryos, a very effective ingredient in the treatment of various diseases was found in rice embryos and named orizinine (vitamin B1). Pectin was extracted to find pectase (enzyme), and colorless crystals of bioactive activity were separated from rice bran and rice embryo oil and named as oranol. The pharmacological action of oranol was hepatic brain function control, UV absorption, antioxidant Actions, peripheral blood vessel dilation, and the like have been confirmed. It is also recognized as a drug for treating symptoms of autonomic ataxia and cosmetic ingredients. GABA has begun to be known in recognition of functional substances.

Brown rice is known to increase nutrients such as vitamins, calcium, minerals, amino acids, enzymes, arabinoxysilane, GABA, etc. when sprouting. It has been reported to have an activating effect, liver function improvement, anti-obesity effect, alcohol metabolism promoting effect, and odor removing effect, and the use of GABA, vitamin B group, minerals, and plant fiber in germination extract as nutritional supplement This is possible, and furthermore, the GABA accumulated in the germination process can be used in food.

In particular, GABA obtained from brown rice germination extract is a functional food material with a physiologically active function to prevent adult diseases, and it is a nutritional supplement containing vitamins and minerals richly used as a beverage such as embryo tea, or mixed into processed foods with powder. Or as granules or tablets, as an additive to food. In addition, the GABA content is increased when the anaerobic treatment of germinated brown rice and green tea during the drying process, grain germination, and ground anatomy of barley seedlings grown for 1 week after barley germination. It has been reported that the increase in large quantities.

Chinese cabbage is very important as winter seasoned kimchi and stocked vegetables, and it is shipped all year round due to the development of cultivation technology and the spread of new varieties and refrigeration facilities. Chinese cabbage contains flavonol, which has antioxidant activities such as hydroxy benzoic acid, hydroxy cinnamic acid, and quercetin, and it is estimated that antioxidant properties are also caused by phenolic compounds and chlorophylls in Chinese cabbage. Chinese cabbage contains carotenoids, vitamin C, and dietary fiber, which have been suggested to have antimutagenic effects on various carcinogens. In addition, cabbage is good for urinating, alcohol is taken out of the body after drinking alcohol is effective in preventing and eliminating hangovers, and eating soup made of cabbage is said to be effective in suppressing emotions and calming nervousness and nervousness.

In today's complex and diverse stressful world, chronic alcohol intake has led to an increase in the number of patients suffering from chronic alcohol consumption. Alcohol is mostly metabolized in the liver, but the liver's ability to process alcohol has limitations, so exceeding this limit will lead to various metabolic disorders. In other words, excessive intake of alcohol increases the ratio of intracellular NADH / NAD ⁺ resulting in disorders of carbohydrates, proteins and lipid metabolism. Increasing toxicity of acetaldehyde can lead to microtubule damage, which in turn can lead to fatty liver and, in severe cases, alcoholic hepatitis or liver cirrhosis. In addition, chronic alcohol intake causes dysfunction of the nervous system, which causes abnormal brain sensations (albeit lower levels of GABA and GAD in the brain) that recognize alcohol effects, resulting in epilepsy, spasticity, Parkinson's disease and schizophrenia. It is known to be closely related to diseases such as the back. The production of GABA is achieved by the decarboxylation of glutamate by glutamate decarboxylase (GAD). Blood GABA levels of alcoholics have been reported to be significantly lower than normal subjects. Alcoholic nervous system diseases have been reported to be related to abnormal levels of GABA, GAD, and GABA receptors in the brain.Alcohol induces neurological diseases by inhibiting the expression of specific areas of the neurotransmitter GABA receptors by 40%. Proposed. As mentioned above, GABA is well known to play an important role, and interest in pharmaceuticals and functional food materials is increasing.

Nakagawa and Onota, a Japanese Ministry of Agriculture, Forestry and Fisheries, found that glutamate decarboxylase (GAD) was automatically activated when water was added to rice bran containing embryos, increasing GABA content. They have developed GABA in embryonic rice bran as a source of functional foods to enhance physiological activity such as lowering blood pressure. Omori et al. Also found that ingestion of GABA-enhanced tea lowered blood pressure in hypertensive rats. This study examined the effects of GABA-enriched cabbage and cabbage root supplementation on fat metabolism and liver function in alcohol-administered rats for the development of functional foods.

γ-aminobutyric acid (GABA) is a nonprotein amino acid that is well known as the major inhibitory neurotransmitter of the central nervous system in animals. GABA is known to be involved in the regulation of many physiological mechanisms to promote brain blood flow and increase oxygen supply in animals to increase brain metabolism. In addition, GABA is involved in the regulation of prolactin and growth hormone secretion, and is known to be effective in lowering blood pressure and pain relief.

As GABA is known to play an important role in the prevention of hypertension and pain relief, interest in GABA as a functional food material as well as GABA as a medicine has recently been increasing. This interest motivated the search for GABA-rich foods and plants and their functional studies, and several studies have been published. For example, Chang et al., Anaerobic treatment of green tea leaves produced in Dawon, Jeju, by the season of red pepper, measured the change in the content of GABA and other major components, and confirmed that the content of GABA is improved. Yun et al also reported that by applying anaerobic treatment to barley germinated during barley malt production, GABA content in malt can be approximately doubled. We confirmed the presence of GABA in the Bulam No. 3 Chinese cabbage leaves. By periodically treating the dilution of chitosan during cabbage cultivation, we found that the GABA content of the Chinese cabbage leaves at the seedling stage and the Chinese cabbage leaves at harvest were approximately doubled. In addition, this study analyzes GABA and amino acids in the leaves and roots of high-altitude Chinese cabbage (Chilsung summer cabbage) as part of the ongoing research into the search for GABA in cabbage and its nutritional biochemical functions. The effects of phosphorus alcohol on the fat metabolism and liver function in rats were investigated.

The following examples are not intended to limit the scope of the present invention, but the present invention will be described in more detail as follows. The analytical reagents and instruments in materials and methods are as follows.

γ-aminobutyric acid was made by Sigma (USA), UV / Vis spectrophotometer was made by Shimadzu (Japan), amino acid analyzer was made by Waters (USA), and other reagents were made by express product. Vision (Korea) product was used. For animal feed, AIN-76 vitamin and mineral mix were made of Teklard (USA) and casein was made of Cottee (Australia). Other feed additives were made of Sigma (USA).

For the analysis of GABA and amino acids in the sample, Chinese cabbage was harvested directly from high-fat summer cabbage (chilseong summer cabbage) grown by fertilizing chitosan fertilizers from organic farms, washed well with water and freeze-dried. To measure the contents of GABA and amino acids in the cabbage leaves and roots, a mixture of methanol: chloroform: water (12: 5: 3) was added to the sample powder ground with liquid nitrogen and mixed. An aqueous layer containing free amino acid and GABA was obtained by centrifugation (12,000xg, 15 min, 4 ^o C). A mixture of chloroform: water (3: 5) was added to the precipitate, and the remaining free amino acid and GABA were extracted secondly, and the supernatant obtained from the first and second centrifugation was combined and lyophilized. Subsequently, it was dissolved in a small amount of water and filtered through a 0.45 μm PVDF filter (Millipore) and analyzed by an automatic amino acid analyzer (AccQ · Tag Amino Acid Analysis System, Waters). Comparing with the content of amino acids and GABA was calculated.

Experimental animals purchased Sprague-Dawley male rats with an average body weight of about 100 g from Daehan Biolink Co., Ltd., Chungju, Korea for 1 week, and then adapted to the first sugar solid food, followed by randomized complete block design. All 6 animals were divided into 4 groups and placed in a stainless steel cage for 30 days. During the experiment, the room temperature was maintained at 23 ± 1 ℃ and the relative humidity was 53 ± 2%. The contrast was illuminated for 12 hours (8: 00-20: 00), and the water and feed were freely fed during the breeding period. .

In the diet and alcohol administration, the composition of the experimental diet is shown in Table 1, and the raw material purified based on the standard amount of the AIN-76 rat specification was used. Chinese cabbage used in the experimental diet was purchased from the organic farming method above, lyophilized and used as a powder (100mesh). During the experimental period (30 days) at 9 am every day, the alcohol group diluted 99% ethyl alcohol with secondary distilled water to make 13% alcohol oral administration (ethanol 3g / kg bw). Instead, second distilled water was orally administered in the same amount.

Table 1.Composition of basal and experimental diet ¹⁾

Ingredients Groups ²⁾ control control + EtOH cabbage leaf + EtOH cabbage root + EtOH Casein 20.0 20.0 20.0 20.0 DL-methionine 0.3 0.3 0.3 0.3 Corn starch 15.0 15.0 10.0 10.0 Cabbage leaf - - 5.0 - carbbage root - - - 5.0 Sucrose 50.0 50.0 50.0 50.0 Fiber 5.0 5.0 5.0 5.0 Corn oil 5.0 5.0 5.0 5.0 AIN mineral mix 3.5 3.5 3.5 3.5 AIN vitamin mix 1.0 1.0 1.0 1.0 Choline bitartrate 0.2 0.2 0.2 0.2

¹⁾ All components are in units of g / 100g diet.

²⁾ In + EtOH groups, 13% (v / v) ethanol (3g / kg bw) was administered for 30 days.

In the treatment of experimental animals, the body weight of the experimental animals during the experimental breeding period was measured in the experimental period 7, 14, 21 days and 30 days, the feed intake was converted to the daily feed intake by measuring the feed residual amount every two days. The final day of experimental breeding was fasted for 12 hours, inhaled anesthesia with ether, opened, and collected by heart blood collection. Blood was left in ice water for about 1 hour and then centrifuged at 1,100 xg for 15 minutes to separate serum and used for the experiment. Soy sauce was extracted immediately after blood collection, washed with physiological saline, and then drained with a filter paper, weighed, and stored in a -70 ° C freezer.

Serum total cholesterol in the lipid and enzyme assays were using a commercially available kit (Youngdong Pharmaceutical), as measured by enzymatic method, HDL-cholesterol is a dextran sulfate-Mg ⁺⁺ precipitation, LDL-cholesterol that are quantitatively precipitated by a precipitation reagent After the supernatant was measured using a kit (Kyoto Pharmaceutical Co. Kyoto, Japan) as when measuring the total cholesterol. Neutral lipids in blood and liver tissues were measured using a commercially available kit (Youngdong Pharmaceutical), and total lipid content in liver tissues was determined using a commercially available kit (Kokusai Pharmaceutical Co., Kobe, Japan) based on the sulfo-phospho-vanillin method. Was used. Serum GOT (glutamic oxaloacetic transaminase), GPT (glutamic pyruvic transaminase) and γ-GTP (γ-gutamyltransferase) enzymes were measured using a commercially available kit (Dong-Pharma).

The statistical results were expressed as Mean ± Standard Deviation (SD), and statistical significance between groups was verified by Duncan's multiple range test at p <0.05 using Statistical Analysis System (SAS) Package. The significance of the amino acid content of cabbage leaf and root was verified by Student's t-test.

The results of GABA and amino acid contents in Chinese cabbage leaf and Chinese cabbage root are shown in Table 2. As shown in Table 2, the amino acid contents of Chinese cabbage leaves were Ser, His, Ala, and GABA, and the total amino acid content including GABA was 59.15 μmol per gram of dry sample, and the ratio of GABA to total amino acids was about 8%. Compared to other amino acids, the GABA content in cabbage roots was much higher, and the GABA content (7.02 μmol / g dry weight) in roots was 1.5 times higher than the GABA content (4.69 μmol / g dry weight) in Chinese cabbage leaves. In addition, GABA accounted for 26.86% of the total amino acid content in the cabbage root. Considering that the cabbage root contains a lot of GABA, we can consider the development of the roots of Chilseong summer cabbage, which are mostly discarded, as a high value-added resource.

Table 2.Free amino acid and GABA compositions in leaves and roots of cabbages ¹⁾

Cabbage leaf (μmol / g dry weight) Cabbage root (μmol / g dry weight) t-test Asp 3.60 (± 0.32) 1.97 (± 0.21) * ²⁾ Ser 10.30 (± 0.52) 1.86 (± 0.05) * Glu 1.51 (± 0.03) 0.74 (± 0.10) * Gly 3.85 (± 0.31) 0.59 (± 0.10) * His 7.61 (± 1.12) 1.27 (± 0.02) * Arg 4.47 (± 0.06) 1.26 (± 0.18) * Thr 3.22 (± 0.05) 1.49 (± 0.02) * Ala 7.81 (± 0.66) 2.58 (± 0.44) * GABA 4.69 (± 0.72) 7.02 (± 1.92) * Pro 1.89 (± 0.36) 1.73 (± 0.62) NS Tyr 0.76 (± 0.06) 0.64 (± 0.08) NS Val 3.49 (± 0.32) 1.54 (± 0.11) * Met 0.33 (± 0.05) 0.24 (± 0.04) NS Lys 1.27 (± 0.11) 0.35 (± 0.06) * Ile 1.82 (± 0.12) 0.91 (± 0.10) * Leu 1.20 (± 0.08) 1.01 (± 0.11) NS Phe 1.33 (± 0.12) 0.94 (± 0.08) NS Total 59.15 (± 5.01) 26.14 (± 4.24) *

¹⁾ Free amino acids containing GABA were extracted from the leaves and roots of cabbages and analyzed. The data represents the mean of three determinations with standard deviation of the mean.

²⁾ Values in the same row are significantly different at p 〈0.05 by Student t-test. NS, nonsignificant.

In the weight change of the experimental animals, the weight loss due to the alcohol administration did not appear at the week after the experimental diet, but from the 2nd week, the alcohol group significantly decreased compared with the non alcohol group (Table 3). Other studies in experimental animals have reported that alcohol intake inhibits the growth of animals because the calorie intake of alcohol reduces dietary intake, loss of body fat and increased energy consumption. In alcoholic patients, dietary intake was significantly reduced, and the absorption of other nutrients was also inhibited, resulting in weight loss due to malnutrition. In this experiment, the alcohol-administered group had a 10% reduction in dietary intake compared to the control group. Addition of 5% cabbage leaf and root did not show any significant weight gain compared to the control group, but it was shown to partially prevent weight loss due to alcohol.

Table 3. Body weight of animals for experimental period

Groups Treatement period (week) 0 One 2 3 4 control 135.9 ± 13.7 ¹⁾ 211.4 ± 18.0 264.9 ± 16.3 ^a2) 309.8 ± 14.3 ^a 342.4 ± 10.1 ^a control + EtOH 138.0 ± 10.0 206.0 ± 16.0 229.6 ± 14.9 ^b 261.7 ± 6.5 ^b 304.0 ± 14.2 ^b Cabbage leaf + EtOH 136.8 ± 12.3 198.8 ± 31.1 245.6 ± 12.4 ^ab 289.2 ± 13.1 ^ab 321.6 ± 10.8 ^ab Cabbage root + EtOH 138.3 ± 8.20 213.0 ± 20.1 259.9 ± 26.9 ^ab 294.4 ± 12.7 ^ab 328.3 ± 13.2 ^ab

¹⁾ The values represent the gram of weight with the standard deviation of the mean of 6 rats per group.

²⁾ Different superscripts in the same columns indicate significant differences (p 〈0.05) among groups by Duncan's multiple range test.

Lipid content in blood and liver tissue is shown in FIG. In the normal diet group, alcohol administration significantly increased the total lipid and triglyceride content and blood triglyceride content in liver tissue compared with the control group. Continued intake of alcohol increases the intracellular NADH / NAD ⁺ ratio resulting in metabolic disorders of carbohydrates, lipids and proteins, inhibits oxidation of fatty acids and increases synthesis, resulting in increased lipid content in blood and liver tissues. It seems to be. The addition of 5% Chinese cabbage lowered the total lipid and triglyceride content in the liver tissues due to the administration of alcohol and also lowered the triglyceride in the blood. Chinese cabbage root supplementation significantly lowered the amount of LDL-cholesterol in the blood. Increasing the amount of LDL-cholesterol induces arteriosclerosis, eventually causing hypertension and hyperlipidemia, coronary heart disease, cerebral infarction and cerebral hemorrhage. In the present study, lowering of total lipid, triglyceride, and LDL-cholesterol in blood and liver tissues should be investigated further. However, the effect is better in the cabbage roots, it can be predicted that GABA, a major component in the roots, may have some effect. In animals, GABA is known to stimulate the blood flow in the brain, increase oxygen supply, and enhance the metabolism of brain cells. Clinically, GABA is used as an improvement drug for stroke sequelae and cerebral atherosclerosis. In addition, the administration of GABA-rich rice embryo extracts to rats significantly reduced the amount of triglycerides in the blood and liver, and it is thought that cabbage can prevent alcohol and heart disease to some extent.

Effects of cabbage diet on lipid concentrations of serum and liver. The error bars show the standard deviation of mean for 6 rats. Values with different letters are significantly different (p 〈0.05) among groups by Duncan's multiple range test. S, serum; HDLc, HDL-cholesterol; LDLc, LDL-cholesterol; TChol, total cholesterol; TG, triglyceride; TL, total lipid. I, control; II, control + EtOH; III, cabbage leaf + EtOH; IV, cabbage root + EtOH.

Chronic alcohol administration in the enzyme concentration of experimental animals increases alcohol oxidation by the microsomal ethanol oxidizing system (MEOS), which produces oxygen radicals such as O ₂ , ^- , OH and H ₂ O ₂ to form lipid peroxides, resulting in hepatocytes. Will cause damage. Abnormalities in hepatocytes increase GOT, GPT, and γ-GTP activity. As shown in Table 4, the blood GOT and γ-GTP activities were significantly higher in the alcohol group than in the control group, suggesting that hepatic tissue damage due to chronic alcohol administration could be estimated. The 5% Chinese cabbage diet was found to lower the γ-GTP activity increased with alcohol to control levels. This suggests that the Chinese cabbage diet may have partial improvement in alcoholic liver damage. As a result of the administration of rice germ extract containing a large amount of GABA to rats, it was found that the concentration of GOT and alkaline peroxidase (ALP) in the blood was lowered. Recently, dietary supplementation of Dong-Gui was found to significantly reduce the level of γ-GTP in the blood due to alcohol administration. GOT and GPT activity in rat serum increased by alcohol administration by supplementing mugwort extract for 6 weeks. It has been reported that administration of Buttercup extract and Prunella vulgaris significantly reduced the increased blood GPT activity due to CCl ₄ administration.

Table 4. Effects of cabbage diet on the levels of several enzymes in plasma

Enzyme Groups control control + EtOH Cabbage leaf + EtOH Cabbage root + EtOH GOT (mU / ml) 76.1 ± 5.3 ^a1) 131.7 ± 6.9 ^b2) 126.1 ± 25.6 ^b 125.3 ± 18.2 ^b GPT (mU / ml) 57.6 ± 1.7 56.3 ± 0.6 55.9 ± 1.2 58.4 ± 2.5 γ-GTP (mU / ml) 10.9 ± 3.7 ^a 27.8 ± 2.8 ^b 12.1 ± 6.5 ^a 11.0 ± 2.8 ^a

¹⁾ The data represents the mean ± SD of 6 rats per group.

²⁾ Different superscripts in the same rows indicate significant differences (p 〈0.05) among groups by Duncan's multiple range test.

Claims (6)

Crops with increased GABA content alleviate the disorders of fat metabolism in liver tissue caused by chronic alcohol intake Effect of GABA in Chinese Cabbage Improves Fat Metabolism and Liver Function in Alcohol Administration The Chinese cabbage root diet of 1 paragraph significantly lowers the amount of LDL-cholesterol in the blood Addition of Chinese Cabbage Leaves and Roots to Lower Total and Neutral Lipids in Blood and Liver Tissues from Alcohol Administration Dietary Supplementation of Cabbage Leaf and Root of Clause 1 Significantly Reduced Blood γ-GTP Activity Increased with Alcohol Effect of the Chinese cabbage diet of 1 paragraph partially improves liver damage caused by alcohol