KR101507704B1 - Composition for detoxification or relieving hangover comprising fermented natural plant as effective component - Google Patents

Abstract

The present invention relates to a composition for liver detoxification or hangover detoxification comprising a fermented product of natural plant material as an active ingredient and is characterized in that the fermented product of soybean, mung bean, parsley, kelp, parsley, radish and cucumber or the fermented product The low-temperature aged material is manufactured using a natural plant material which is safe and easily obtainable and low-cost raw material so as to replace the liver detoxification and hangover composition using existing expensive medicinal plants. As the effect of eliminating healing and hangover is excellent, it can be provided as a composition useful for consumers' health and safe liver detoxification or hangover.

Description

TECHNICAL FIELD The present invention relates to a composition for detoxification or hangover of a liver comprising a fermented product of natural plant material as an active ingredient,

The present invention relates to a composition for liver detoxification or hangover detoxification comprising a fermented product of natural plant material as an active ingredient, and more particularly to a composition for liver detoxification or hangover decomposition which contains a fermented product of natural plant material as an active ingredient. Mixed fermentation products; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low-temperature aged product as an active ingredient, and a health food.

The intake of acute and chronic alcohol causes damage to various organs such as liver, kidney and brain, and induces alcoholic hepatitis and cirrhosis in particular, resulting in death. When a small amount of alcohol is consumed in a normal state, ethanol entering the liver is converted to acetate by the action of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in the cell matrix, It is excreted through the circulation system outside the hepatocytes. In particular, acetaldehyde, the first metabolite of ethanol, is highly reactive and toxic, which is the main cause of alcoholic liver disorder. It causes not only hangover but also inhibits the function of mitochondria, an intracellular energy producing organ. Cirrhosis of the liver and heart and brain function is inhibited. Acetaldehyde also promotes collagen synthesis in myofibroblast, causing hepatic fibrosis and metamorphosis of hepatocytes, and reacting with macromolecules in vivo to form adducts.

The degradation pathway of alcohol in the liver is divided into three types of enzymes: alcohol dehydrogenase (ADH pathway), microsomal ethanol oxidizing system (MEOS), and catalase pathway. The clinically important pathway is the oxidation of alcohol to acetal via both acetaldehyde and ADH system and MEOS system. The AHD system is involved in alcohol metabolism when the tissue alcohol concentration is low, and the MEOS system is involved in alcohol metabolism. It is called alcohol metabolism. In order to prevent hangover liver damage, the consumed alcohol should be rapidly degraded to acetate. To do so, the biosynthesis of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) and the activity of these enzymes should be enhanced. In addition, in order to prevent the poisoning due to acute and chronic alcohol ingestion, it is possible to reduce the hangover and tissue damage by promoting the metabolism of the consumed alcohol and inhibiting or eliminating the production of ROS involved in tissue damage. Various medicines and health functional foods have been developed and sold domestically in order to alleviate hangovers caused by alcohol and to prevent or prevent long-term damage. However, since the mechanism of organ damage by alcohol consumption is very diverse and complex, It is not enough to acknowledge.

On the other hand, Korean Patent No. 1150885 discloses red ginseng roots, red ginseng roots, ungerminated soybean extract and mixed compositions of hangover, but it is also possible to use the roots of the present invention as the active ingredients of liver plant detoxification or hangover The composition has not been disclosed.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a composition for liver detoxification or hangover detoxification derived from natural materials which is safe and easily obtainable from human body, And cucumber powder were mixed at a certain ratio and fermented and matured at a low temperature to be orally administered to rats. As a result, it was found that the present invention was completed by confirming the prevention and healing effect on liver tissue damage caused by alcohol consumption and the excellent effect on the elimination of hangover Respectively.

To achieve the above object, the present invention provides a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the fermented product or low-temperature aged product as an active ingredient.

The present invention also relates to a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, powdered pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low temperature aged product as an active ingredient.

The fermented product of the fermented product of soybean, mung bean, parsley, kelp, parsley, radish, and cucumber or the fermented product at low temperature of the present invention is safe to replace the liver detoxification and hangover composition using existing expensive medicinal plants In addition, it is manufactured using a natural plant material which is easily available and inexpensive raw material. The composition is excellent in protection of liver tissue, healing and hangover, so that it is useful for consumers' health and safe for liver detoxification or hangover .

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow chart illustrating a process for preparing the composition of the present invention. FIG.
FIG. 2 is a graph showing the results of evaluating the degree of damage of liver-induced liver tissue of rats orally administered with the composition of the present invention. (A) glutathione content, (B) lipid peroxide content, (C) triglyceride content, (D) alanine aminotransferase (ALT), (E) aspartate aminotransferase : Ethanol control, NFME: mixed extracts added 5.0%, FME: fermented mixed extracts added 5.0%, FLME: low temperature aged mixed extracts added 5.0%.
FIG. 3 is a graph showing the results of evaluation of the degree of activity of an alcohol metabolism-related enzyme in rats orally administered with the composition of the present invention. (A) Alcohol dehydrogenase activity, (B) mitochondria-aldehyde dehydrogenase activity, (C) cytoplasmic-aldehyde dehydrogenase activity, (D) cytoplasmic cytochrome p450 2E1 activity, NC: normal group, EN: ethanol control group, NFME: FME: fermented mixed extract of 5.0%, FLME: low temperature fermented mixed extract of 5.0% added dietary group.
4 is a graph showing the results of measurement of changes in blood alcohol concentration in rats orally administered with the composition of the present invention. NC: normal group, EN: ethanol control group, NFME: mixed extract 5.0% added diet group, FME: fermented mixed extract 5.0% added diet group, FLME: low temperature aged mixed extract 5.0% diet group.

In order to achieve the object of the present invention, the present invention provides a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the fermented product or low-temperature aged product as an active ingredient.

The mixed fermented product according to one embodiment of the present invention may be prepared by mixing 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, 3 to 7 parts by weight of kelp powder, 3 to 7 parts by weight of powdered powder, 3 to 7 parts by weight of powder-free powder and 3 to 7 parts by weight of cucumber powder, fermenting by adding fermenting microorganism, preferably 100 parts by weight of the mixture, Mixed with 70 parts by weight of powder, 5 parts by weight of mungbean powder, 5 parts by weight of powdered mung bean, 5 parts by weight of tallow powder, 5 parts by weight of powdered pomegranate powder, 5 parts by weight of powderless powder and 5 parts by weight of cucumber powder, But is not limited thereto.

The mixed fermentation product is more specifically,

(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;

(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And

(c) fermenting the mixture with the fermenting microorganism at a temperature of 35 to 40 ° C for 2 to 3 days under a humidity of 70 to 90%, but is not limited thereto.

The low-temperature fermented product according to an embodiment of the present invention may be prepared by mixing 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, , 3 to 7 parts by weight of powdered powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder, fermenting the mixture by adding a fermenting microorganism, and then aging at low temperature, The mixture was mixed with 70 parts by weight of soybean powder, 5 parts by weight of mung bean powder, 5 parts by weight of parsley powder, 5 parts by weight of seaweed powder, 5 parts by weight of powdered pear powder, 5 parts by weight of no powder and 5 parts by weight of cucumber powder based on 100 parts by weight of the mixture Fermenting microorganism may be added and fermented, followed by low-temperature aging, but the present invention is not limited thereto.

More specifically, the low-temperature aged product of the above-

(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;

(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And

(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature But it is not limited thereto.

The mixed fermented product or the low-temperature aged extract according to an embodiment of the present invention may be prepared by enzymatic extraction, hot water extraction or alcoholic extraction of the low-temperature aged product of the mixed fermented product or the mixed fermented product, but is not limited thereto. Enzyme extraction may be, but is not limited to, alkalase extraction. Further, the mixed fermented product or the low-temperature aged product may be prepared by subjecting the low-temperature aged product of the mixed fermented product or the mixed fermented product to enzyme extraction, hot water extraction or alcohol extraction, and then combining the respective extracts.

In a composition for liver detoxification or hangover resolution according to an embodiment of the present invention, the fermenting microorganism is selected from the group consisting of Bacillus sp. licheniformis , Bacillus pumilus), Bacillus Sono alkylene sheath (Bacillus sonorensis , and Bacillus subtilis . Preferably, it may be Bacillus licheniformis . However, the present invention is not limited thereto.

In clinical practice, the degree of hepatic tissue damage can be determined by measuring the activities of glutathione, lipid peroxidation, triglyceride, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in liver tissue. Or rats were sacrificed for 4 weeks. After 12 hours of fasting, ethanol was orally administered to measure the activity. As a result, rats fed with the composition of the present invention were found to have liver tissue similar to that of normal rats without oral administration of ethanol (FIG. 2), and the activities of mitochondrial-aldehyde dehydrogenase and cytoplasmic-aldehyde dehydrogenase, which are alcohol metabolism-related enzymes, were excellent (FIG. 3). In addition, it was confirmed that the concentration of ethanol in the blood of the rats fed with the composition of the present invention was markedly decreased after 1 hour (FIG. 4). Therefore, the composition of the present invention can be used for prevention and improvement of hepatitis and cirrhosis, but is not limited thereto.

In addition,

(a) drying and then pulverizing the washed soybean, mung bean, parsley, kelp, parsley, radish and cucumber;

(b) sterilizing the soybean, mung bean, parsley, kelp, parsley, radish, and cucumber powder and mixing with the fermenting microorganism; And

(c) fermenting the mixture. The present invention also provides a method for preparing a composition for liver detoxification or hangover decomposition.

The method of the present invention for preparing liver-decrypting or hangover-

(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;

(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And

(c) fermenting the mixture with the fermenting microorganism at a temperature of 35 to 40 ° C for 2 to 3 days under a humidity of 70 to 90%.

In addition,

(a) drying and then pulverizing the washed soybean, mung bean, parsley, kelp, parsley, radish and cucumber;

(b) sterilizing the soybean, mung bean, parsley, kelp, parsley, radish, and cucumber powder and mixing with the fermenting microorganism; And

(c) fermenting the mixture, and then aging the mixture at a low temperature. The present invention also provides a method for preparing a composition for liver detoxification or hangover decomposition.

The method of the present invention for preparing liver-decrypting or hangover-

(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;

(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And

(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature Step < / RTI >

The present invention also relates to a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, powdered pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low temperature aged product as an active ingredient.

When the fermented product of the present invention or an extract of the fermented product at low temperature or the fermented product or the fermented product at low temperature is used as a food additive, the fermented product or the fermented product or the fermented product of the fermented product The extract of the low-temperature aged product can be directly added or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the fermentation product or the low-temperature aged product or extract of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. As a specific example, the fermented product, the low-temperature aged product or the extract may be used to produce a processed food having good storability while being modified by utilizing characteristics of agricultural products, livestock products or aquatic products. Such processed foods include, for example, confectionery, drinks, liquor, fermented foods, canned foods, milk processed foods, meat processed foods, noodles and the like. The sweets include biscuits, pies, breads, candies, jellies, gums, cereals (including dinner utensils such as cereal flakes). Drinks include carbonated beverages, functional ionic beverages, juices (such as apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.) and sikhye. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned products include canned products (for example, tuna, mackerel, sandwiches, canned fish, etc.), canned products (canned beef, pork, chicken and turkey canned products) and canned products (corn, peach and pineapple canned products). Milk processed foods include cheese, butter, yogurt and the like. Meat processed foods include pork cutlet, beef cutlet, chicken cutlet, sausage, sweet and sour pork, nuggets, nubucki, and the like. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.

In addition, functional foods, health foods or health supplements can be produced by using the above-mentioned fermented products or low-temperature aged products or extracts. A functional food, a health food or a health supplement food means a food which provides a biomodulating function including a physiologically active ingredient in addition to a nutrition function, and the fermented product or the low-temperature aged product or extract of the present invention has a function Since the above-mentioned various natural products are contained as an effective ingredient, they can be used for the production of functional foods, health foods or health supplements.

Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples. However, the following examples and preparative examples are merely illustrative of the present invention, and the present invention is not limited to the following examples and preparative examples.

Manufacturing example  1: liver detox or hangover mixed for small Fermentation product  And mixing Fermented  Production of low temperature aged water

The soybean ( Glycine max L.) was grown in Hwacheon, Gangwon province, and mung bean ( Phaseolus radiatus, Vigna radita) is in Gimhae, Gyeongnam, kelp (Laminaria japonica ) were purchased from Wando County, Chonnam Province. Puerariae Radix , Oenanthe javanica ), cucumber ( Cucumis sativus L.) and radish ( Raphanus sativus L.) purchased and used domestic organic products. As a strain, Bacillus ricinifolium ( Bacillus sp. licheniformis strains were used. Alcalase (2.4 L (2.4 AU / g)) was purchased from Sigma, and the reagents used for the efficacy analysis were standard limited reagents.

The low-temperature aged product of the liver-detoxification or hangover-reducing small-scale fermentation product and the mixed-fermentation aged product of the present invention was produced through the flow chart of FIG.

The first step is a drying and pulverizing step. First, soy beans, kelp, mung bean, parsley, parsley, radish, and cucumber were screened and washed, and each material was rapidly lyophilized to minimize loss of nutrients. In order to increase the extraction of useful components and the promotion of fermentation, each of the dried products was first pulverized with a crusher and then a 120 mesh powder was prepared using a fine pulverizer.

The second step is a step of blending the compounding, sterilizing and fermenting microorganisms. It is a step of mixing the soybean, kelp, mung bean, parsley, bean, radish and cucumber powder having various physiological activities with a proper ratio 7: 0.5: 0.5: 0.5: 0.5: 0.5: 0.5, w / w) and sterilized at 80 ° C for 40 minutes. The fermentation was used for Bacillus subtilis (B. licheniformis) demonstrated the usefulness, was used to uniformly inoculated with this cell was washed by incubation at 37 ~ 40 ℃ for 24 hours in a powder, a certain amount of time so that the solid fermentation is fully yirwoji Distilled water (5% of the powder weight) was added and then molded to a predetermined size and mixed with the powder mixture.

In the third step, the powder mixture inoculated with the fermenting microorganisms was fermented at 70 to 90% of humidity and at 35 to 40 ° C for 2 to 3 days so that sufficient fermentation could be achieved. Fermented mixture '. The fermentation mixture was aged at a temperature of 10 to 20 ° C for 7 days under a humidity of 40 to 60%, and the composition thus obtained was used as a 'low-temperature aged product of the fermentation mixture'.

The fourth step is an extraction and drying step. In order to reduce the risk of corruption, the solid fermentation product fermented and fermented at low temperature was dried at 60 ° C for 12 to 24 hours so as to have a moisture content of 5% or less in an infrared drying apparatus. The extraction was carried out in three steps of extraction, hot water extraction and alcohol extraction so that the functional oil component of the composition could be sufficiently extracted. 10 times water was added to the low-temperature fermented product or mixed fermented product of the third step, and the mixture was ultrasonically extracted at 50 ° C. Then, the enzyme was extracted for 60 minutes by the addition of alkalase, and enzyme was removed for 10 minutes at 95 ° C And a 10-fold amount of water was added to the residue, and the mixture was subjected to hot extraction at 80 ° C for 9 hours to prepare a second extract. The residue was added with 10-fold amount of 70% alcohol at 60 ° C A third extract was prepared.

The fifth step is a step of mixing, concentrating and formulating. The supernatant obtained by centrifuging the first, second and third extracts of the fourth step is inactivated with enzyme, mixed, filtered, The mixture was concentrated under reduced pressure to obtain a brick, thereby obtaining a fermented mixed composition extract. The hot-air dried product of the fourth step and the fermented mixed composition extract concentrate were mixed at a ratio of 95: 5 (w / v) and dried at 60 ° C for 12 to 24 hours to have a water content of 6% or less. Respectively.

Experimental Method

1. Experimental animals and Dietary  pharmacy

Experimental animals were fed with Sprague-Dawly SPF / VAF outbred rats (Orient) with an average body weight of 150 ± 10 g at 6 weeks of age. The experimental diets were prepared as shown in Table 1, Bricks. 1: 1: 1: 1: 1: 1: 1) was added to the test animals (NC), the ethanol control (EN) and the soybean flour powder (kelp, mung bean, : 5.0% addition of fermented mixed powder extract, 5.0% fermented mixed powder extract and 5.0% addition of fermented aged mixed powder extract (mixture of fermented microbial extract and non fermented mixture) (Total 5 groups) were fed with 7 rats for 4 weeks. For 4 weeks, 50% ethanol solution was orally administered in 5 g / kg of body weight once daily for 2 days in fasting state. After 12 hours, the animals were treated. The temperature and humidity were adjusted to 60 ± 5% and 23 ± 2 ℃, and the dark and light cycles were set at 12 - hour intervals, and water and feed intake were freely consumed.

Feed composition by experimental treatments Dietary composition Experimental group ¹⁾ NC EN NFME FME FLME Solid feed ^{2 )} 100 100 95.0 95.0 95.0 Mixed extract - - 5.0 - - Fermented mixed extract - - - 5.0 - Fermentation ripening mixed extract - - - - 5.0

¹⁾ NC: Normal group, EN: Ethanol control group, NFME: Mixed extract 5.0% added diet group, FME: Fermented mixed extract 5.0% added diet group, FLME: Fermentation, Low temperature aged mixed extract 5.0%

^{2) 5} L79 diet of PMI Nutrition (LLC, PO Box 19798, Brentwood MO 63144, USA). Ingredients: crude protein 18%, crude fat 5%, crude fiber 5%, ash 8%.

To determine the degree of liver damage, the content of glutathione (GSH) in the liver was calculated by subtracting the amount of thiophenol produced by adding 2-nitrobenzoic acid to the liver homogenate of the extracted animal Respectively.

The content of lipid peroxide (LPO) was determined by adding thiobarbituric acid (TBA) solution to the liver tissue homogenate and adding thiobarbituric acid reactant (TBA -active substance was measured at 532 nm and the content was calculated using molecular extinction coefficient (ε = 1.5 × 10 ⁵ M ^-1 cm ^-1 ).

The lipid content of liver tissues was measured by Folch's method (Folch J et al., J Biol Chem 226: 497-509, 1957) and used as a lipid measurement sample. The total lipid content was determined by the method of Frings and Dunn RT, Am J Clin Path 53: 89-91, 1970).

The activity of alanine aminotransferase (ALT) was measured using a kit reagent (ASAN PHARMACEUTICAL, Korea). The activity was expressed as Karmen unit per 1 mL of serum. Protein content of liver tissue was measured by using bovine serum albumin (BSA) as a standard solution.

The degree of enzymatic activity of hepatic tissues is determined by xanthine oxidoreductase (XOD), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione S- The activity of glutathione S-transferase (GST), alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and cytochrome P450 2E1 (Cytochrome P450 2E1) The circles were homogenized by adding 4 times ice cold 0.25 M sugar solution to a fixed amount of hepatic tissues of the extracted animals and then using a postmitochondrial fraction (PMF) and a mitochondrial fraction centrifuged at 10,000 rpm for 20 minutes.

In the measurement of the activity of xanthine oxidoreductase, the total type activity was measured in the presence of nicotinic acid amide, and the content of uric acid produced in the absence of nicotinic acid amide (O type) activity was measured at 292 nm .

The superoxide dismutase activity was measured at an absorbance of 560 nm, and the activity was expressed as the protein content (unit) of 50% inhibition of autoxidation of hematoxylin in the absence of the enzyme. The catalase activity was measured at 240 nm in the content of hydrogen peroxide which was decomposed during the reaction.

The activity of glutathione peroxidase decreases the content of nicotinamide adenine dinucleotide phosphate (NADPH), which is used to convert glutathione reductase to glutathione by reducing reduced glutathione, which is consumed to reduce the substrate hydrogen peroxide, The activity of glutathione S-transferase was determined by measuring the activity of thioether produced by incorporating 1-chloro-2,4-dinitroezene (CDNB) and reduced glutathione, Absorbance at 340 nm.

The activity of the aldehyde dehydrogenase was determined by measuring the absorbance at 340 nm of the content of nicotinamide adenine dinucleotide produced during the reaction of acetaldehyde and nicotinic acid amide as substrates, and the activity was determined by measuring the activity of nicotinamide adenine dinucleotide In terms of nmole.

The activity of cytochrome P450 2E1 (CYP2E1) was determined by the addition of aniline (Sigma Aldrich, USA) and nicotinamide adenine dinucleotide to the enzyme source, and the activity of p-aminophenol / hr < / RTI > in nmole.

Weight, dietary, and drinking water intake were measured at regular intervals throughout the day. Dietary efficiency was calculated by dividing daily gain by daily dietary intake.

The rats fed the diet for 4 weeks were allowed to drink water freely. After fasting for 12 hours, blood was drawn from the abdominal aorta under ether anesthesia. The liver was perfused with ice-cold physiological saline, the organs were removed, Respectively. The collected blood was coagulated at room temperature, centrifuged at 2500 rpm for 20 minutes at 4 ° C, serum was separated, and stored at -70 ° C for analysis.

Example  1: Mixed fermented product for liver detoxification or hangover treatment of the present invention and mixed fermented aged product of  Alcoholic Liver damage  Prevention and healing effects

FIG. 2 shows the result of the oral administration of 50% ethanol equivalent to 5 g / kg of body weight to the experimental animals grown for 4 weeks on the prevention of alcoholic liver damage and the healing effect.

As a result of the analysis of glutathione content, which is known to detoxify lipid peroxidation in the body, the ethanol control group treated with 50% ethanol was increased compared to the normal group and the mixed extract, mixed fermented product and mixed fermented product added diet, In the case of mixed fermented product, the content of glutathione was lower than that of the normal group, and the fermented fermented product remained in the normal group level (FIG. 2A).

The content of lipid peroxidation, which is known to be an indicator of cell membrane damage caused by reactive oxygen species, was rapidly increased by the acute treatment of ethanol compared to that of the normal group, whereas the mixed extract of natural plant material, In the fermented aged group, the amount of lipid peroxidation, which was significantly increased by ethanol, recovered to the normal level. Especially, the mixed fermented aged diet - supplemented group was decreased compared with the normal group and all other extract supplemented diet group. Therefore, it was concluded that the various nutrients contained in natural plant material mixed extract, mixed fermented product and mixed fermented aged product have a function of preventing or alleviating acute toxicity of ethanol, resulting in a low content of liver lipid peroxidation. Particularly, in the case of the mixed fermented water diet group, the content of lipid peroxidation and triglyceride was remarkably decreased while the normal value of glutathione was maintained. This indicates that various useful substances produced by fermentation and aging can prevent and recover ethanolic liver toxicity (Fig. 2B).

When ethanol is consumed, neutral lipids are not released into the liver tissue and accumulate and cause fatty liver. Analysis of triglyceride content of liver tissues showed that the content of triglyceride of liver tissue was significantly increased in the ethanol control group (EN) compared to that of the normal group, while that of natural plant material mixed extract (NFME), mixed fermented product (FME) In the fermented (FLME) diet group, 22.8%, 37.2% and 38.1% of the ethanol group were decreased, respectively. These results suggest that the increase in liver triglyceride content in the ethanol control group is due to the acute toxicity of ethanol. Based on the above experimental results, the mixed fermentation ages of the natural plant material mixture showed the acute toxicity of ethanol (Fig. 2C).

The activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) is increased by hepatic tissue injury and is widely used as an index of liver tissue damage. Analysis of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) showed that the ethanol control group, which was negative control group, showed a significant increase compared to the normal group, (Figs. 2D and 2E).

These results suggest that the activity of serum alanine aminotransferase and aspartate aminotransferase and the content of hepatic lipid peroxidation and triglyceride in the ethanol control group were significantly increased by the toxicity of acute ethanol. It was confirmed that the toxicity of acute ethanol was prevented or reduced by the diet, and no damage of the liver tissue occurred.

Example  2: liver detoxification of the present invention or mixture of hangover Fermentation product  And mixed fermentation Aged  Evaluation of enzymatic activity of liver tissue

In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of xanthine oxidoreductase in liver tissues, xanthine oxidase, superoxide dismutase, glutathione peroxidase, The activities of catalase and glutathione estersparation, alcohol dehydrogenase, mitochondria-aldehyde dehydrogenase, cytoplasmic-aldehyde dehydrogenase and cytoplasmic cytochrome p450 2E1 were measured.

Xanthine oxidase is the normal physiological condition under the nicotinic acid amide xanthine dehydrogenase, using as an electron acceptor: using the present as (xanthine dehydrogenase XOR type D), but between textural molecule instead of nicotinic acid amide damage the oxygen (O ₂₎ as an electron acceptor And converted to xanthine oxidase (XOR type O) to produce reactive oxygen species (ROS) such as superoxide and hydrogen peroxide. After 4 weeks, 50% ethanol equivalent to 5g / kg of body weight was orally administered to experimental animals, and the activity of xanthine oxidase activity of liver tissue was measured. In the case of total type of xanthine oxidase (XOR total type), the ethanol control group was decreased by acute administration compared with the normal group, whereas the mixed extract of natural plant material, mixed fermented product and mixed fermented product were significantly increased Especially, the fermented fermented diet group showed higher activity than the normal group. On the other hand, the activity of xanthine oxidase (XOR O type) of xanthine oxidase was increased by the acute administration of ethanol compared with that of the normal group, but decreased to below the normal group in the mixed extract of natural plant material, mixed fermented product and mixed fermented water (Table 2).

Determination of xanthine oxidoreductase activity of liver tissue of mixed fermented product and mixed fermented aged product for hepatotoxicity or hangover injury of the present invention
Experimental group Xanthine oxidoreductase (nmol / mg protein) Total Type
(Total type) Oh type
(O type) Rate ^{1 )}
(O / T,%)  Normal group 155.79 ± 6.89 ²⁾ 27.80 + - 4.70 17.99 ± 3.84  Ethanol control group 114.72 + - 7.07 38.79 ± 2.92 33.97 + - 3.80  Mixed extract 140.06 + - 6.71 22.90 ± 4.59 16.35 ± 3.24  Mixed fermentation product 152.19 + - 5.65 22.29 ± 2.14 14.66 + 1.50  Mixed fermented aged material 157.10 + - 4.77 17.39 ± 2.76 11.10 ± 1.96

¹⁾ (O type / total type) X 100 = ratio (%).

²⁾ Mean value and standard deviation of 7 measurements.

In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of the active oxygen scavenging enzymes of hepatic tissues, superoxide dismutase, glutathione peroxidase, Catalase and glutathione S-transferase activities were measured, and the results are shown in Table 3. (O ^{2 -} ) in the group of natural plant material mixed extracts, mixed fermented products and mixed fermented aged diets were converted into hydrogen peroxide which was weakly toxic, and again hydrogen peroxide was added to water to make a detoxified superoxide dismutase, glutathione peroxidase , Catalase activity was generally increased in comparison with the ethanol control group, which was a negative control, and it was confirmed that the activity was significantly increased in the mixed fermented water diet group. It is glutathione S-transferase that converts glutathione into a weakly toxic ethanolic fat and makes it non-toxic when making active lipid peroxidation lipid peroxidation. Glutathione S-transferase activity of the mixed fermented product was slightly decreased in the ethanol control group, but no significant change was observed in all experimental groups (Table 3).

As a result, the mixed fermentation aged mixture of the natural plant material helps increase the activity of the antioxidant enzymes in the liver, and this phenomenon also appears in the administration of acute ethanol, so that it is confirmed that the liver damage caused by ethanol is prevented and alleviated I could.

Measuring the activity of the active oxygen scavenging enzymes in the liver tissues of the mixed fermented product and the mixed fermentation aged product for hepatotoxicity or hangover injury of the present invention Experimental group Superoxide-dismutase ^{1 )} Glutathione peroxidase ^{2 )} Catalase ^{3 )} Glutathione-S-transferase ^{4 )} Normal group 34.22 ± 3.20 ⁵⁾  98.29 ± 7.99  257.09 ± 19.10  468.85 +/- 29.53 Ethanol control group  18.87 + - 4.26  70.61 + - 7.28  105.89 ± 13.55  441.14 ± 29.82 Mixed extract  52.55 + 4.18  85.36 ± 8.93  228.57 ± 19.52  478.00 ± 25.70 Mixed fermentation product  60.93 + - 5.98  88.05 + - 9.98  226.48 ± 17.28  461.43 ± 24.10 Mixed fermented aged material  65.07 + - 7.36  86.47 ± 9.68  251.43 + - 16.00  466.29 ± 30.04

^1-4) Superoxide dismutase unit: unit (content of protein that inhibits 50% of autoxidation of hematoxylin), catalase unit: nmole / protein mg, glutathione peroxidase unit: nmole / protein mg, glutathione S-transferase: nmole / protein mg.

⁵⁾ Mean value and standard deviation of 7 measurements.

In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of the alcohol metabolism-related enzymes in liver tissues, the alcohol dehydrogenase, mitochondrial-aldehyde dehydrogenase, cytoplasmic-aldehyde dehydrogenase The results of evaluating the activity of the enzyme and cytoplasmic cytochrome p450 2E1 are shown in FIG.

The activity of alcohol dehydrogenase, which is oxidized to acetaldehyde and converted to acetic acid by acetaldehyde dehydrogenation and finally converted to acetyl CoA to produce energy, (Fig. 3A), whereas the mitochondrial fraction of hepatic tissue, the major metabolic enzyme of acetaldehyde, and cytoplasmic aldehyde dehydrogenase were significantly increased compared to the ethanol control (Fig. 3B and Fig. 3C). On the other hand, the activity of cytochrome P450 2E1 was significantly reduced in the natural plant material mixed extract, mixed fermented product, and mixed fermented aged diet (FIG. 3D).

As a result, the mixed fermentation aging of the mixture of natural plant material shows the activity of high acetaldehyde dehydrogenase, which rapidly metabolizes acetaldehyde, which is a strong toxic substance, compared with other extracts. Therefore, It was confirmed that the prevention or alleviation effect was excellent.

Example  3: Evaluation of Dietary Efficacy and Blood Alcohol Concentration Change of Mixed Fermented Products and Mixed Fermentation Agents for Liver Detoxification or Hanging Habits of the Present Invention

Table 4 shows the results of observing weight gain, dietary intake and dietary efficiency during the 4 weeks experimental diet. The daily average weight gain from the differences in body weight and initial body weight was 4.54 g in the normal group and 3.96 g in the ethanol control group, which was decreased by 11.89%. On the other hand, mixed extracts of natural plant material, mixed fermented product and mixed fermented product showed higher dietary intake than ethanol - only control. The intake of drinking water tended to be somewhat higher in the ethanol - treated group. Weight gain and dietary efficiency were decreased in the ethanol control group, but the fermented mixed extract group had a higher value than the other experimental control groups, indicating that the phenomenon of physiological fluctuation due to alcohol consumption was low.

The dietary intake, drinking water intake, body weight gain and dietary efficiency of the mixed fermented product and mixed fermented aged product for hepatotoxicity or hangover injury of the present invention Experimental group Dietary intake
(g / day) Drinking water intake
(g / day) Weight gain
(g / day) Dietary efficiency ^{1 )}  Normal group 4.54 ± 0.22 ²⁾ 45.71 + - 4.27 5.07 ± 1.15 1.12 ± 0.27  Ethanol control group 3.96 + 0.31 56.43 + - 5.56 3.19 ± 0.89 0.82 ± 0.27  Mixed extract 4.14 + 0.33 52.14 + - 6.36 3.73 ± 0.48 0.90 + - 0.10  Mixed fermentation product 4.59 ± 0.33 50.71 + - 4.50 4.21 + - 0.42 0.92 + 0.13  Mixed fermented aged material 4.49 ± 0.33 51.14 + - 4.02 4.59 ± 0.73 1.02 + - 0.16

¹⁾ Dietary efficiency: daily gain / daily dietary intake

²⁾ Mean value and standard deviation of 7 measurements

To investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the change of the ethanol concentration with time, 50% ethanol was added to 2 g per kg of body weight And the changes of ethanol concentration in blood were examined. The ethanol control group reached its peak concentration, then gradually decreased and decreased to about 40% after 4 hours. On the other hand, in the case of the mixed extract of natural plant material, the concentration of ethanol in the blood gradually decreased after 30 minutes, and the ethanol concentration of the mixed fermented product and mixed fermented aged food group of the present invention was remarkably decreased. In particular, the change in the blood alcohol concentration with time was found to be large in the mixed fermentation aged product, and this result is considered to be the result of the above experiment in which the CAT activity, which is an auxiliary agent of alcohol metabolism, is high (FIG.

Thus, the activity of liver tissue alcohol dehydrogenase in the dietary group with various sample extracts was lower than that of the ethanol control group, which is a negative control group, because the activity of catalase, which is an enzyme involved in the conversion of ethanol into acetaldehyde, It can be interpreted that the activity of the aldehyde dehydrogenase which converts the aldehyde to acetic acid is increased and the active oxygen generating enzyme which is involved in the liver tissue damage is inhibited and the activity of the active oxygen scavenging enzyme is increased.

Claims (8)

Low temperature aging of mixed fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pupae powder, no powder and cucumber powder; Or an extract of low-temperature aged product of said mixed fermented product as an active ingredient, wherein the low-temperature aged product of said mixed fermented product is produced by the following steps:
(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature step. delete delete [3] The composition of claim 1, wherein the low-temperature aged extract of the mixed fermented product is prepared by enzymatic extraction, hot water extraction or alcoholic extraction of the low-temperature aged product of the mixed fermented product. delete delete According to claim 1, wherein said fermenting microorganism is selected from the group consisting of Bacillus piece you formate miss (Bacillus licheniformis), Bacillus pumil Ruth (Bacillus pumilus), Bacillus Sono alkylene sheath (Bacillus sonorensis) and Bacillus subtilis (Bacillus subtilis) Wherein the composition is at least one kind of liver detox or hangover. Low temperature aging of mixed fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pupae powder, no powder and cucumber powder; Or a low-temperature aged product of said mixed fermented product as an active ingredient, characterized in that the low-temperature aged product of said mixed fermented product is produced by the following steps: food:
(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature step.

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