Composition containing fermented kiwi fruit for preventing or improving alcoholic liver damage

Abstract

The present invention relates to a composition for preventing and ameliorating alcoholic liver damage and a composition for relieving hangovers, which comprises a kiwi fermented product obtained by fermenting kiwi fruit with lactic acid bacteria as an active ingredient. According to the present invention, the kiwi fermented product of the present invention promotes alcohol metabolism in the liver, thereby reducing blood ethanol and aldehyde concentrations as alcohol metabolism is activated, and suppresses fat accumulation and the production of inflammatory cytokines in the liver, as well as cholesterol and triglyceride accumulation, thereby preventing or ameliorating alcoholic liver damage and relieving various hangover symptoms.

Description

[Technical Field]

[0001] This invention was made under project number S3260875 with support from the Ministry of Small and Medium Enterprises and Startups of Korea, and the project management specialist is the Small and Medium Enterprise Technology Information Promotion Agency, the research project name is "Regional Specialized Industry Development + (R&D)", the research topic name is "Development and commercialization of health functional food material for suppressing alcoholic liver damage from fermented gold kiwifruit using novel lactic acid bacteria", the project execution organization is "Bitec Corporation", and the research period is from April 1, 2022 to March 31, 2023.

[0002] The present invention relates to a composition for preventing or ameliorating alcoholic liver damage, which comprises a kiwi fermented product, and more specifically, to a composition for preventing or ameliorating alcoholic liver damage, which comprises a kiwi fermented product fermented with lactic acid bacteria as an active ingredient. [Background technology]

[0003] Ninety percent of alcohol absorbed in the intestine is metabolized within the body, while the remaining 10% is excreted unmetabolized through urine, breath, sweat, etc. Most of the absorbed alcohol is metabolized in the liver. Although some alcohol is also metabolized in the brain, pancreas, and stomach, the amount is significantly smaller than that of the liver. Because the liver is the primary organ responsible for alcohol metabolism, chronic alcohol consumption is known to frequently result in liver injury. While there are various causes of liver injury, persistent, unregulated inflammation can lead to cumulative liver fibrosis, ultimately increasing the risk of progression to cirrhosis and liver cancer.

[0004] Alcoholic liver injury is generally caused by increased hepatic fat accumulation due to alcohol consumption and increased oxidative stress and inflammatory responses due to alcohol metabolism. Alcohol-induced hepatic fat accumulation shares some similarities with the process of hepatic fat accumulation due to overnutrition, in that excessive alcohol consumption causes an oversupply of fat to the liver, which promotes the breakdown of free fatty acids in adipose tissue and increases the influx of free fatty acids into the liver. However, it differs in that alcohol metabolism reduces nicotinamide adenine dinucleotide (NAD) to reduced NAD (NADH), which increases the NADH / NAD ratio, resulting in an imbalance between carbohydrate and fat metabolism, decreased gluconeogenesis, and increased fatty acid synthesis. Alcohol-induced hepatic fat accumulation leads to alcoholic fatty liver, a condition that generally resolves with abstinence. However, because it is largely asymptomatic, it also requires periodic management. Oxidative stress and inflammatory responses caused by alcohol metabolism are triggered by lipid peroxidation, which occurs when a significant amount of reactive oxygen species produced during alcohol metabolism damages blood cells and cell membranes. Persistent inflammatory responses ultimately lead to the production of reactive aldehydes, which promote liver fibrosis. Acetaldehyde, a highly reactive substance produced during alcohol metabolism, depletes glutathione in the liver by forming various adducts with proteins or DNA, selectively reducing glutathione in mitochondria and reducing antioxidant capacity. It also promotes the production of inflammatory cytokines, including TNF-α, which induces liver cell toxicity and ultimately leads to liver damage.

[0005] Currently, as the incidence of alcoholic liver disease in Korea is relatively increasing, its importance among liver diseases is also increasing. It has been reported that 18.6% of actual liver cirrhosis patients show a relationship with alcohol. Therefore, it is necessary to prevent and improve alcoholic liver damage to prevent the development of various liver diseases.

[0006] Kiwi fruit is the fruit of a dioecious, deciduous vine in the Actinidiaceae family (Acinidiaceae) and Actinidia genus, primarily grown in temperate regions. The fruit is covered in brown hairs, resembling the New Zealand bird "kiwi," hence the name. In Korea, it is also called "yandare" or "jamdare." Kiwi fruit contains phenolic compounds known to contribute not only to the taste and flavor of food but also to various physiological activities. It is rich in vitamins C and E, and contains large amounts of minerals such as folic acid, potassium, calcium, and phosphorus. It also contains bioactive substances beneficial to health, such as chlorophyll and carotenoids. Kiwi fruit contains a wide variety of bioactive substances, and diverse research using kiwi fruit continues. Specifically, Korean Patent No. 1081910 relates to a cosmetic composition containing kiwi extract for improving skin tone and preventing skin aging. It discloses that the kiwi extract effectively inhibits skin glycation, thereby improving skin tone and reducing wrinkles. Korean Patent No. 2027798 relates to an antioxidant composition containing gold kiwi lactic acid bacteria fermentation product as an active ingredient. It discloses that gold kiwi lactic acid bacteria fermentation product, prepared by fermentation with a Lactobacillus plantarum strain culture medium, has significantly increased total phenol and flavonoid contents compared to unfermented gold kiwi, resulting in excellent antioxidant activity. Furthermore, actinidin, a protein-degrading enzyme, is effective in promoting digestion, making kiwi fruit known as a fruit beneficial for stomach health. However, the prior art does not mention the effect of kiwi fermentation product in improving alcoholic liver damage.

[0007] Therefore, the present inventors have conducted extensive research and efforts to overcome the problems of the prior art. As a result, they have discovered that a composition for preventing or ameliorating alcoholic liver damage, which contains as an active ingredient a kiwi fermented product obtained by fermenting kiwi fruit with lactic acid bacteria, promotes alcohol metabolism in the liver, thereby reducing blood ethanol and aldehyde concentrations and suppressing fat accumulation and the production of inflammatory cytokines in the liver, as well as cholesterol and triglyceride accumulation, thereby preventing or ameliorating alcoholic liver damage, and have completed the present invention. Summary of the Invention [Problem to be solved by the invention]

[0008] Therefore, the main object of the present invention is to provide a composition for preventing or ameliorating alcoholic liver damage, which contains kiwi fermented product, and which can prevent or ameliorate alcoholic liver damage by suppressing fat accumulation and the production of inflammatory cytokines in the liver, promoting alcohol metabolism in the liver to reduce blood ethanol and aldehyde concentrations, and suppressing cholesterol and triglyceride accumulation.

[0009] Another object of the present invention is to provide a composition for relieving hangovers, which contains a fermented kiwi fruit, and which can relieve hangovers by promoting alcohol metabolism in the liver and thereby reducing blood ethanol and aldehyde concentrations as alcohol metabolism is activated.

[0010] Another object of the present invention is to provide beverages and foods containing the hangover relief composition. [Means for solving the problem]

[0011] According to one aspect of the present invention, there is provided a composition for preventing or ameliorating alcoholic liver damage, comprising, as an active ingredient, a kiwi fermented product obtained by fermenting kiwi with lactic acid bacteria.

[0012] The term "prevention or amelioration of alcoholic liver damage" in the present invention means preventing or ameliorating liver damage by inhibiting or suppressing factors that can damage the liver caused by alcohol consumption, and ultimately preventing liver disease caused by liver damage. Specifically, it means preventing or ameliorating alcoholic liver damage by promoting alcohol metabolism in the liver, thereby reducing blood ethanol and aldehyde concentrations as alcohol metabolism becomes more active, suppressing fat accumulation and the production of inflammatory cytokines in the liver, as well as suppressing the accumulation of cholesterol and triglycerides.

[0013] The kiwifruit in the present invention is a kiwifruit of the genus Actinidia chinensis (gold kiwifruit), and examples of the kiwifruit varieties include Jessie Gold (Actinidia chinensis Planch var. chinensis 'Jecy Gold'), Halla Gold (Actinidia chinensis Planch var. chinensis 'Halla Gold'), Haegum (Actinidia chinensis Planch var. chinensis 'Haegum'), Zespri Gold (Actinidia chinensis Planch var. chinensis 'Hort16A'), Zespri SunGold (Actinidia chinensis Planch var. chinensis 'Zesy002'), Zespri Zesy003 (Actinidia chinensis Planch var. chinensis 'Zesy003'), and Zespri Zesy003. chinensis 'Zesh004'), Zespri ZESH004 (Actinidia chinensis Planch var. chinensis 'Zesh004'), Consorzio Dori Europe Dori (Actinidia chinensis Planch var. chinensis 'AC1536'), and Jingold (Actinidia chinensis Planch 'Jintao').

[0014] In the composition for preventing or improving alcoholic liver damage of the present invention, the lactic acid bacteria is a kiwi-derived lactic acid bacteria, and the kiwi-derived lactic acid bacteria is one or more of Lactococcus lactis VI-01 KCTC14351BP and Lactobacillus paracasei VI-02 KCTC14352BP.

[0015] The present inventors isolated lactic acid bacteria from strains isolated from kiwifruit and decoded the genome sequences of the selected strains by 16s rRNA analysis. The strains selected by genome sequencing were named "Lactococcus lactis VI-01" and "Lactobacillus paracasei VI-02" and deposited at the Korea Comprehensive Life Science Center (KCTC) of the Korea Institute for Bioscience and Biotechnology (KCTC) on November 3, 2020, with accession numbers KCTC14351BP and KCTC14352BP, respectively.

[0016] In the composition for preventing or improving alcoholic liver damage of the present invention, the lactic acid bacteria are lactic acid bacteria generally used for fermentation in the art, and are preferably Lactobacillus paracasei, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus gasseri, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus The present invention is characterized in that the present invention contains one or more strains selected from the group consisting of Lactobacillus reuteri, Lactobacillus rhamnosus, and Lactobacillus salivarius, and more preferably one or more strains selected from the group consisting of Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus helveticus.

[0017] The kiwi fermented product of the present invention can be obtained by culturing a bacterial strain in a first medium (seed culture) to produce a first culture product (seed culture), inoculating the first culture product into a second medium and culturing it (mass culture) to produce a second culture product (mass culture), centrifuging the second culture product, removing the supernatant, and mixing the resulting culture with kiwi puree and fermenting it. More specifically, the first culture product (seed culture) can be produced by inoculating the bacterial strain into each medium at 0.01 to 0.5% and culturing it at 35 to 37°C for 18 to 28 hours, while the second culture product (mass culture) can be produced by inoculating the bacterial strain into each medium at 0.01 to 1% and culturing it at 35 to 37°C for 7 to 10 hours. The mixing ratio of kiwi puree to culture is 7.5 to 8.5:2.5 to 1.5, preferably 8:2.

[0018] In the composition for preventing or ameliorating alcoholic liver damage of the present invention, the kiwi fermented product is contained in an amount of 30 to 80 wt %, preferably 30 to 60 wt %, based on the total weight of the composition. If the amount of the kiwi fermented product is less than 30 wt %, it is difficult to exhibit a sufficient effect of preventing or ameliorating alcoholic liver damage, while if the amount of the kiwi fermented product is more than 80 wt %, the effect of preventing or ameliorating alcoholic liver damage relative to the amount contained is insufficient, which is undesirable from an economical perspective.

[0019] The composition for preventing or ameliorating alcoholic liver damage of the present invention is characterized in that the composition is a functional health food composition.

[0020] In the composition for preventing or ameliorating alcoholic liver damage of the present invention, the functional health food composition may have a dosage form that is convenient for ingestion and use, preferably one or more dosage forms selected from the group consisting of capsules, tablets, powders, granules, liquids, pills, pieces, pastes, syrups, gels, beverages, jellies, and bars. To be prepared in such dosage forms, the composition may further contain conventional excipients, stabilizers, thickeners, etc. Furthermore, the composition may further contain food additives, and their suitability as "food additives" is determined in accordance with the specifications and standards for the relevant item based on the general provisions and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety, unless otherwise specified.

[0021] When the functional health food composition of the present invention is formulated into a powder, the kiwi fermented product can be rapidly frozen at -180°C to -195°C, preferably at -195°C, using a liquid nitrogen freezer (LNF), and then freeze-dried at -35°C to -45°C, preferably at -40°C, to obtain a dried material, which can be pulverized to form the formulation.

[0022] The composition for preventing or ameliorating alcoholic liver damage of the present invention is characterized in that the composition is a food composition.

[0023] In the composition for preventing or improving alcoholic liver damage of the present invention, the food composition can be formulated into a food product that can be used with fermented kiwi fruit as a main ingredient, and is preferably in the form of one or more selected from the group consisting of dairy products, beverages, sauces, jams, and confectioneries.

[0024] According to one experimental example of the present invention, the fermented kiwi fruit of the present invention reduced increased inflammatory cytokines in alcohol-induced mice (EtOH) (see Experimental Example 2), reduced increased ethanol and aldehydes in alcohol-induced mice (EtOH) (see Experimental Example 3), and reduced increased cholesterol and triglycerides in alcohol-induced mice (EtOH) (see Experimental Example 4). These results suggest that the fermented kiwi fruit of the present invention can prevent or ameliorate liver damage by inhibiting or suppressing factors that cause liver damage caused by alcohol.

[0025] According to another aspect of the present invention, there is provided a composition for relieving hangovers, comprising, as an active ingredient, a kiwi fermented product obtained by fermenting kiwi fruit with lactic acid bacteria.

[0026] The term "hangover relief" in the present invention refers to the alleviation of physical hangover symptoms such as headache, fatigue, muscle pain, bloodshot eyes, thirst, general malaise, abdominal distension, vomiting, upset stomach, and decreased cognitive function, which are induced by ingested alcohol and its metabolic products, aldehydes. Specifically, it refers to the elimination of hangover symptoms induced by ethanol and aldehydes by promoting alcohol metabolism in the liver and reducing blood ethanol and aldehyde concentrations as alcohol metabolism becomes more active.

[0027] In another aspect, the present invention relates to a hangover relief beverage or food product, including a hangover relief composition containing, as an active ingredient, a kiwi fermented product obtained by fermenting kiwi fruit with lactic acid bacteria.

[0028] The food product according to the present invention may be in one or more dosage forms selected from the group consisting of powder, granules, tablets, capsules and pills, in addition to beverages. [Effects of the Invention]

[0029] As described above, the composition containing the kiwifruit fermented product according to the present invention promotes alcohol metabolism in the liver, activates alcohol metabolism, reduces blood ethanol and aldehyde concentrations, suppresses fat accumulation and the production of inflammatory cytokines in the liver, and suppresses cholesterol and triglyceride accumulation, thereby preventing or improving alcoholic liver damage and eliminating various hangover symptoms.

Brief Description of Drawings

[0030] [Figure 1] It is a figure showing the Gram staining of the deposited strain according to the present invention. [Figure 2] It is a figure showing the formation of a clear zone by the decomposition of calcium carbonate of the deposited strain according to the present invention. [Figure 3] It is a figure observing the morphology of the deposited strain Lactococcus Lactis VI-01 KCTC14351BP according to the present invention. [Figure 4] It is a figure observing the morphology of the deposited strain Lactobacillus paracasei VI-02 KCTC14352BP according to the present invention. [Figure 5] It is a phylogenetic tree of the deposited strain Lactococcus Lactis VI-01 KCTC14351BP according to the present invention. [Figure 6] It is a phylogenetic tree of the deposited strain Lactobacillus paracasei VI-02 KCTC14352BP according to the present invention. [Figure 7] It is the result of measuring the body weight change when kiwifruit alcohol was treated on mice induced by alcohol (Normal: untreated with alcohol, EtOH: treated with alcohol, FGL: treated with alcohol + kiwifruit fermented product (50 mg / kg), FGM: treated with alcohol + kiwifruit fermented product (125 mg / kg), FGH: treated with alcohol + kiwifruit fermented product (250 mg / kg)). [Figure 8a]These are the results of examining blood cytokines when alcohol-induced mice were treated with fermented kiwi fruit (TNF-α, Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + fermented kiwi fruit (50 mg / kg), FGM: alcohol treatment + fermented kiwi fruit (125 mg / kg), FGH: alcohol treatment + fermented kiwi fruit (250 mg / kg)). [Figure 8b] These are the results of examining blood cytokines when alcohol-induced mice were treated with fermented kiwi fruit (IL-1β, Normal; no alcohol treatment, EtOH; alcohol treatment, FGL: alcohol treatment + fermented kiwi fruit (50 mg / kg), FGM: alcohol treatment + fermented kiwi fruit (125 mg / kg), FGH: alcohol treatment + fermented kiwi fruit (250 mg / kg)). [Figure 8c] These are the results of examining blood cytokines when alcohol-induced mice were treated with fermented kiwi fruit (IL-6, Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + fermented kiwi fruit (50 mg / kg), FGM: alcohol treatment + fermented kiwi fruit (125 mg / kg), FGH: alcohol treatment + fermented kiwi fruit (250 mg / kg)). [Figure 9a] These are the results of examining the blood ethanol levels when mice with alcohol-induced hypertension were treated with kiwi fermentation product (Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + kiwi fermentation product (50 mg / kg), FGM: alcohol treatment + kiwi fermentation product (125 mg / kg), FGH: alcohol treatment + kiwi fermentation product (250 mg / kg)). [Figure 9b] These are the results of examining blood aldehyde levels when alcohol-induced mice were treated with fermented kiwi fruit (Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + fermented kiwi fruit (50 mg / kg), FGM: alcohol treatment + fermented kiwi fruit (125 mg / kg), FGH: alcohol treatment + fermented kiwi fruit (250 mg / kg)). [Figure 10a]These are the results of examining blood cholesterol levels when alcohol-induced mice were treated with kiwi fermentation product (Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + kiwi fermentation product (50 mg / kg), FGM: alcohol treatment + kiwi fermentation product (125 mg / kg), FGH: alcohol treatment + kiwi fermentation product (250 mg / kg)). [Figure 10b] These are the results of examining blood triglycerides when alcohol-induced mice were treated with fermented kiwi fruit (Normal: no alcohol treatment, EtOH: alcohol treatment, FGL: alcohol treatment + fermented kiwi fruit (50mg / kg), FGM: alcohol treatment + fermented kiwi fruit (125mg / kg), FGH: alcohol treatment + fermented kiwi fruit (250mg / kg)). DETAILED DESCRIPTION OF THE INVENTION

[0031] The present invention will be described in more detail below with reference to examples. These examples are for the purpose of illustrating the present invention, and therefore the scope of the present invention should not be construed as being limited by these examples.

[0032] Preparation example 1: Isolation and identification of microorganisms 1) Separation source and sample pretreatment Fresh gold kiwi fruit and puree were used to isolate lactic acid bacteria from gold kiwi fruit. Fresh gold kiwi fruit were Zespri Gold Kiwi fruit imported from New Zealand, purchased at a mart in September 2020, washed with triple distilled water to remove foreign matter, and then chopped finely, including the skin. Gold kiwi puree was purchased from Namyang Frozen Foods Co., Ltd. in May 2020 and stored at -20°C. It was completely dissolved at room temperature and mixed uniformly before use.

[0033] 2) Natural fermentation Chopped gold kiwi fruit and puree were fermented under salted and de-fatted conditions. For the salted condition, each sample was salted for 3 hours with 8% solar salt added, and then mixed with a sterilized 1% fructooligosaccharide solution. For the de-fatted condition, each sample was mixed with a sterilized 2% de-fatted and 1% fructooligosaccharide solution. The pH of all the mixed samples was adjusted to 6.0 or higher using sodium hydroxide solution, and they were fermented in an incubator at 37°C for approximately 3 days.

[0034] 3) Selection and identification of lactic acid bacteria The fermented sample was mixed homogeneously and then a portion of the fermentation broth was inoculated onto BCP plate count agar (EIKEN, Japan) and cultured at 37°C for 48 hours. After the medium turned yellow, colonies with similar phenotypes were plated onto MRS agar (Difco, USA) for pure isolation. The isolated strains were identified by colony characteristics and microscopic observation (×1000), and only strains that showed Gram-positive activity were initially selected (Figure 1). The strains were then inoculated onto MRS agar containing 1% calcium carbonate (DAEJUNG, Korea) and cultured at 37°C for 48 hours. Two strains that formed large clear zones due to the decomposition of calcium carbonate by organic acids were finally selected (Figure 2).

[0035] The two isolated strains were arbitrarily designated VI-01 and VI-02, and stocks were prepared using 30% glycerol and stored at -80°C. To confirm the biochemical properties of the two strains, such as their sugar degradation ability and their ability to decompose arginine and esculin, the API 50 CHL kit (Biomerieux, France) was used to identify medium discoloration using the apiweb program (http: / / apiweb.biomerieux.com).

[0036] Biofact Inc. was commissioned to perform PCR and sequencing analysis using 27F and 1492R primers for 16S rRNA analysis. The nucleotide sequences of each strain were compared for homology with the 16S ribosomal RNA gene sequences registered in GenBank using NCBI's blast program, and alignments were performed using the ClustalX 2.1 program using the neighbor-joining method. Phylogenetic trees were constructed using the bootstrap NJ tree method in ClustalX 2.1 and confirmed with the NJplot program.

[0037] 4) Genome sequencing Strain VI-01 was analyzed for its 16S rRNA gene and its 1,414 bp nucleotide sequence was confirmed to be 100% identical to Lactococcus lactis NBRC 100933. Therefore, strain VI-01 was designated Lactococcus lactis VI-01 (SEQ ID NO: 1).

[0038] Strain VI-02 was analyzed for its 1,441 bp 16S rRNA gene sequence, which showed 99% identity with Lactobacillus paracasei strain R094. Therefore, strain VI-02 was designated Lactobacillus paracasei VI-02 (SEQ ID NO: 2).

[0039] 5) Identification by sugar utilization survey The isolated strains were identified by a sugar utilization test using the API 50 CHL kit. Strain VI-01 was found to be 98.4% similar to Lactococcus lactis, and strain VI-02 was found to be 99.6% similar to Lactobacillus paracasei.

[0040] Preparatory Example 2: Biochemical and morphological characteristics of the strain 1)Biochemical properties The biochemical characteristics of the isolated strains were measured using the API 50 CHL kit. Colonies cultured in pure culture on MRS agar medium were diluted to the appropriate concentration and inoculated into the API 50 CHL medium. The mixture was then cultured at 37°C for 24-48 hours, and the discoloration of the inoculated medium was observed.

[0041] The Lc. lactis VI-01 strain utilized 19 of the 49 sugars, including galactose, D-glucose, D-fructose, D-mannose, mannitol, maltose, and lactose, but was unable to utilize sorbitol or xylitol (Table 1).

[0042] [Table 1]

[0043] The L. paracasei VI-02 strain utilized 22 of the 49 sugars, including galactose, D-glucose, D-fructose, D-mannose, mannitol, and sorbitol, but was unable to utilize lactose and xylitol (Table 2).

[0044] [Table 2]

[0045] 2) Morphological characteristics Morphological characteristics were examined by examining the Colinii morphology and color of the strains cultured on MRS agar medium, and by observing the strain morphology under a microscope.

[0046] The colonies of L. lactis VI-01 are small, rounded, and shiny white, measuring 0.5 to 1.5 mm. Microscopic observation of the strain's morphology confirmed that it was a cocci (Figure 3).

[0047] The colonies of L. paracasei strain VI-02 are round, convex, and shiny white or cream-colored, measuring 1.5 to 2.5 mm. Microscopic observation revealed that the colonies formed long chains (Figure 4).

[0048] Preparation example 3: Strain deposit After 16S RNA sequence analysis and morphological characterization, and finally strain confirmation, the strain was deposited at the National Biological Resource Center and given a deposit number (Table 3).

[0049] [Table 3]

[0050] Example: Production of kiwi fermented product using lactic acid bacteria 1) Kiwi stock Although various varieties of kiwi can be used, in the present invention, kiwi of the genus Actinidia chinensis (gold kiwi) was used.

[0051] 2) Pre-treatment process (production of kiwi puree) The kiwi fruits are washed several times and selected for use as raw materials. The selection criteria are that the fruit is free of blemishes, rotten or contaminated with bacteria, and has a sugar content of 13% Brix or higher (if it does not exceed this, it will undergo a post-ripening process). The selected fruit is peeled and the flesh is shredded.

[0052] 3) Grinding and seed separation process (production of kiwi puree) For grinding and seed separation, the shredded pulp is put into a centrifugal grinder and ground, and the seeds are removed by filtering through a sieve, and then the mixture is stirred to make it homogenous.

[0053] 4) Lactic acid bacteria cultivation process (seed culture - tank culture - centrifugation) 4-1) Lactic acid bacteria species The lactic acid bacteria used to produce the kiwi fermented product were the Lactobacillus lactis VI-01 KCTC14351BP and Lactobacillus paracasei VI-02 KCTC14352BP strains isolated in previous experiments, mixed with three other lactobacillus species: Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus helveticus. The three lactobacillus species were from Sacco.

[0054] 4-2) Culture process 4-2-1) Seed culture Seed culture was performed in MRS Broth. Specifically, each strain was inoculated into the respective medium at 0.1% and cultured at 37°C for 24 hours to obtain the primary culture product (strain). Sterilization was performed at 121°C for 15 minutes.

[0055] 4-2-2) Mass culture The first culture was used for large-scale cultivation. The first culture was inoculated into MRS medium and cultured at 37°C for 9 hours to obtain the second culture. Sterilization was carried out at 121°C for 20 minutes.

[0056] 4-2-3) Centrifugation and concentration The cultured lactic acid bacteria are centrifuged to concentrate the culture medium and the lactic acid bacteria strains.

[0057] 5) Mixing The raw material (kiwi puree) and lactic acid bacteria culture solution were mixed in a ratio of 8:2. The specific composition and inoculation amount are shown in Table 4 below.

[0058] [Table 4]

[0059] 6) Fermentation The mixed raw materials are cultured at 37°C for 8 to 12 hours.

[0060] 7) Powderization For use in the experiment, the prepared fermented product was quick frozen and freeze-dried, and then crushed into powder to prepare kiwi fermented product powder.

[0061] Preparation example 4: Preparation of animal model The experimental animals used in this study were 40 8-week-old ICR male mice (8 mice / group) purchased from Damul Science (Daejeon, Korea) and allowed to adapt to the housing environment for one week. After adaptation, the mice were weighed and divided into five groups of 8 mice each using a randomized block design: normal (Normal), oral administration of EtOH 5g / kg BW (EtOH), 5g / kg EtOH and 50mg / kg BW of Fermented Gold Kiwi Powder (FGL), 5g / kg EtOH and 125mg / kg BW of Fermented Gold Kiwi Powder (FGM), and 5g / kg EtOH and 250mg / kg BW of Fermented Gold Kiwi Powder (FGH). For the first two weeks of the experiment, the normal and EtOH groups were orally administered DW only, while the Fermented Gold Kiwi Powder group was orally administered each concentration of the sample once daily. For the next two weeks, the normal group received only DW twice a day, while the EtOH group received both ethanol and DW. The fermented gold kiwi powder groups received additional oral administration of 50 mg, 125 mg, or 250 mg of powder per kg of body weight dissolved in ethanol for two weeks. The temperature of the experimental animal room was 22 ± 2°C, the relative humidity was 50 ± 10%, the ventilation rate was 10–20 times per hour, and the lighting cycle was adjusted to 12-hour intervals. During the experiment, mice were provided with water and food ad libitum. All animal experimental procedures were approved by the Chonbuk National University Animal Experiment Ethics Committee (JBNU 2022-095).

[0062] Experimental Example 1: Alcohol-induced weight measurement in mice Since alcoholic liver damage causes weight gain, the effect of fermented kiwi fruit in improving alcoholic liver damage was confirmed by measuring the weight of alcohol-induced mice.

[0063] Specifically, the animals were divided into groups according to the randomized block design, and their weights were measured and recorded. The weights of the animals were measured individually once a week, and the average values ​​were calculated and shown.

[0064] As a result, as can be seen in Figure 7, the weight gain in alcohol-induced mice (EtOH) that were not treated with kiwi fermented food was reduced when treated with kiwi fermented food at different concentrations (FGL (50 mg / kg), FGM (125 mg / kg), FGH (250 mg / kg)).

[0065] Experimental Example 2: Confirmation of blood cytokines in mice induced by alcohol The effect of fermented kiwi fruit in improving alcoholic liver damage was confirmed by examining the blood cytokine levels in mice induced by alcohol.

[0066] Specifically, measurements were performed using an ELISA kit capable of measuring each cytokine. The serum and cytokine antibody cocktail were added to a 96-well plate containing each cytokine antibody, and the plate was incubated at room temperature for 1 hour, followed by washing with wash buffer. After 10 minutes of incubation, TMB buffer was added to each well, and the reaction was terminated by adding stop buffer. The absorbance was measured at 450 nm using a microplate reader. The concentration of each cytokine was calculated from a curve generated using the standard solution included in the ELSIA kit (Abcam, London, UK).

[0067] As can be seen from Figures 8a to 8c, the inflammatory cytokines that increased in alcohol-induced mice (EtOH) without treatment with kiwi fermented food were reduced when treated with kiwi fermented food at various concentrations, with IL-6 in particular being significantly reduced.

[0068] Experimental Example 3: Confirmation of ethanol and aldehydes in the blood of mice induced by alcohol The effect of fermented kiwi fruit in improving alcoholic liver damage was confirmed by examining the blood ethanol and aldehyde concentrations in the mice induced by alcohol.

[0069] Specifically, an ethanol assay kit (Abcam, AB65343) was used to measure blood alcohol concentration. The serum was mixed with the reaction mix and incubated at 37°C for 30 minutes, followed by an additional hour at room temperature. The reaction was performed in the dark during the entire reaction. After the reaction was completed, the absorbance was measured at 570 nm using a microplate reader.

[0070] To measure blood aldehyde concentrations, an aldehyde quantification assay kit (ABCAM, AB112113) was used. After mixing the obtained serum with the 2x yellow mixture, the reaction was allowed to proceed at room temperature for 30 to 60 minutes in the dark, and the absorbance was then measured at 550 nm using a microplate reader.

[0071] As a result, as can be seen from Figures 9a and 9b, the increased ethanol and aldehyde levels in alcohol-induced mice (EtOH) that were not treated with kiwi fermented food were reduced when treated with kiwi fermented food at different concentrations.

[0072] Experimental Example 4: Confirmation of blood cholesterol and neutral fat levels in mice induced by alcohol The effects of fermented kiwi fruit on improving alcoholic liver damage were confirmed by examining the blood cholesterol and triglyceride levels in mice induced by alcohol.

[0073] Specifically, blood samples were collected after the experiment and centrifuged at 1,500 rpm for 20 minutes at 4°C to obtain serum, which was used in this experiment. Total cholesterol and triglyceride levels in the blood were measured using an assay kit manufactured by BIOMAX. The concentrations of each TC and TG were calculated from a curve generated using the standard solution included in the kit.

[0074] As a result, as can be seen from Figures 10a and 10b, the increased cholesterol and triglycerides in alcohol-induced mice (EtOH) that were not treated with kiwi fermented food were reduced when treated with kiwi fermented food at various concentrations. [Sequence List Free Text]

[0075] SEQ ID NO:1: Lactococcus Lactis VI-01 SEQ ID NO:2: Lactobacillus paracasei VI-02

[0076] [Contract information] Contracting organization name: Korea Center for Biological Resources (KCTC) Accession number: KCTC14351BP Entrustment date: November 3, 2020 Contracting organization name: Korea Center for Biological Resources (KCTC) Accession number: KCTC14352BP Entrustment date: November 3, 2020 TIFF2025533788000006.tif205157 JPEG2025533788000007.jpg203157

Claims

1. A composition for preventing or improving alcoholic liver damage, comprising as an active ingredient a kiwi fermented product obtained by fermenting kiwi with lactic acid bacteria, wherein the lactic acid bacteria is one or more of Lactococcus lactis VI-01 KCTC14351BP and Lactobacillus paracasei VI-02 KCTC14352BP, which are lactic acid bacteria derived from kiwi.

2. 2. The composition for preventing or improving alcoholic liver damage according to claim 1, wherein the lactic acid bacteria comprises one or more strains selected from the group consisting of Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus helveticus.

3. The composition for preventing or improving alcoholic liver damage according to claim 1, wherein the fermented kiwi fruit is contained in an amount of 30 to 80% by weight based on the total weight of the composition.

4. The composition for preventing or improving alcoholic liver damage according to claim 1, wherein the composition is a functional health food composition.

5. The composition for preventing or improving alcoholic liver damage according to claim 4, characterized in that the health functional food composition is in one or more dosage forms selected from the group consisting of capsules, tablets, powders, granules, liquids, pills, pieces, pastes, syrups, gels, beverages, jellies, and bars.

6. The composition for preventing or ameliorating alcoholic liver damage according to claim 1, wherein the composition is a food composition.

7. The composition for preventing or improving alcoholic liver damage according to claim 6, wherein the food composition is in the form of one or more selected from the group consisting of dairy products, beverages, sauces, jams, and confectioneries.

8. A composition for relieving hangovers, comprising as an active ingredient a kiwi fermented product obtained by fermenting kiwi fruit with lactic acid bacteria, wherein the lactic acid bacteria is one or more of Lactococcus lactis VI-01 KCTC14351BP and Lactobacillus paracasei VI-02 KCTC14352BP, which are lactic acid bacteria derived from kiwi fruit.

9. A hangover relief drink comprising the composition of claim 8.

10. A hangover relief food comprising the composition according to claim 8.

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