KR20200099115A - Functional food composition for preventing or improving non-alcoholic steatohepatitis comprising fermented ginseng - Google Patents

Abstract

The present invention relates to a health functional food composition for preventing or alleviating non-alcoholic steatohepatitis containing fermented ginseng. More specifically, the present invention relates to a health functional food composition for preventing or alleviating non-alcoholic steatohepatitis, wherein the composition alleviates inflammation in the liver, inhibits liver fibrosis, and improves factors related to the effect of reducing lipid accumulation in the liver.

Description

Functional food composition for preventing or improving non-alcoholic steatohepatitis comprising fermented ginseng}

The present invention relates to a health functional food composition for preventing or improving nonalcoholic steatohepatitis containing fermented ginseng, and more specifically, the composition improves inflammation in the liver, inhibits liver fibrosis, and improves factors related to the effect of reducing lipid accumulation in the liver. It relates to a functional food composition for preventing or improving non-alcoholic steatohepatitis.

The mortality rate of liver disease in Korea is very high at 235 people per 100,000 people, and is the number one cause of death in their 40s (411/10,000 people), the second cause of death in their 50s (724/10,000 people), and the third cause of death in their 30s. Liver disease, accounting for (10/10,000), is a major cause of death in the middle-aged Korean population.

Among the liver diseases, fatty liver refers to a phenomenon in which triglycerides that do not exist in normal cells are abnormally deposited in liver cells. About 5% of normal liver is composed of adipose tissue, and triglycerides, fatty acids, phospholipids, cholesterol and cholesterol esters are the main components of fat, but once fatty liver occurs, most of the components are replaced by triglycerides and the amount of triglycerides is reduced. Fatty liver is diagnosed if it is more than 5% of the weight of the liver.

When the fatty liver deteriorates and the fat mass in the hepatocyte becomes large, the important components of the cells including the nucleus are pushed to one side and the function of the hepatocyte decreases, and the expanded hepatocytes due to the accumulated fat in the cells press the microvessels and lymph glands between the liver cells. Thus, the circulation of blood and lymph fluid in the liver is impaired. In this case, the liver cells cannot properly receive oxygen and nutrients, and liver function is degraded.

Non-alcoholic fatty liver disease (NAFLD) is defined as a case where fatty acids accumulate more than 5% in the parenchymal cells of the liver in the form of triglycerides, not liver damage caused by alcohol. Pathologically, it is classified into simple steatosis and steatohepatitis with inflammation.In particular, non-alcoholic steatosis causes hepatocellular degeneration/necrosis as fat accumulates in hepatocytes, resulting in inflammation and Since liver fibrosis develops into cirrhosis and liver cancer, it is recognized as a serious disease worldwide.

However, there are no drugs or treatments that have proven effective in the treatment of these non-alcoholic steatohepatitis. Therefore, most of the underlying diseases that cause steatohepatitis are treated, and lifestyle changes such as exercise, diet, and weight loss are required.In addition, diabetes and insulin resistance treatment, treatment of hyperlipidemia, prevention and treatment of oxidative stress, and other liver protection It is being treated using drugs.

In the prior art Korean Patent Registration No. 10-1837173, it is described for a pharmaceutical composition for the prevention and treatment of non-alcoholic fatty liver comprising a ginseng seed oil extract extracted by a compression extraction method.

However, as for the efficacy of the ginseng seed oil extract, which is discarded during processing and cultivation of ginseng, not ginseng, and employs a different material from fermented ginseng using the probiotics and pectinase enzyme of the present invention, and prevents non-alcoholic steatohepatitis Or, it does not describe the use of improvement.

Accordingly, the present inventors prepared ginseng fermented ginseng using probiotics and pectinase enzymes to improve inflammation in the liver, inhibit liver fibrosis, and reduce lipid accumulation in the liver, and prevent or improve non-alcoholic steatohepatitis without side effects. The present invention was completed by developing a composition for use.

Korean Patent Registration No. 10-1837173 (2017.12.08)

Therefore, the main object of the present invention is a health functional food for the prevention and improvement of non-alcoholic steatohepatitis containing fermented ginseng that improves inflammation-related factors in the liver, such as improving liver inflammation, inhibiting liver fibrosis, and reducing lipid accumulation in the liver. To provide.

The present invention is to solve the technical problem by providing a health functional food composition for preventing or improving non-alcoholic steatohepatitis containing fermented ginseng as an active ingredient.

The present invention is to solve the technical problem by providing a health functional food composition for preventing or improving non-alcoholic steatohepatitis, characterized in that the fermented ginseng contains 100 to 400 mg/kg based on oral administration of a mouse.

The present invention is to solve the technical problem by providing a health functional food composition for preventing or improving non-alcoholic steatohepatitis, characterized in that the composition improves inflammation in the liver, inhibits liver fibrosis, and inhibits lipid accumulation in the liver.

The present invention, fermented ginseng, the first step of selecting and washing ginseng; A second step of cutting the washed ginseng into 01 to 1 cm; Purified water was mixed with the cut ginseng, and a mixture of enzyme strains in which a pectinase enzyme and a Saccharomyces servazzii strain were mixed at a weight ratio of 2:3 was inoculated at 30-60° C. for 5 days. A third step of fermenting to prepare a fermented ginseng broth; And a fourth step of preparing fermented ginseng by concentrating, drying and pulverizing the ginseng fermentation broth, and the prepared fermented ginseng is ginsenoside-Rg1 18.71 mg/g, Re 43.51 mg/g, Rh1(s)+Rg2 (s) 40.45 mg/g, Rb2 18.73 mg/g, Rd 10.23 mg/g, Rg3(s) 4.36 mg/g and Compound K 23.32 mg/g by providing a method for producing fermented ginseng, characterized in that it contains I want to solve the technical problem.

As described above, according to the present invention, fermented ginseng has been shown to have the effect of improving inflammation in the liver, inhibiting liver fibrosis, and reducing lipid accumulation in the liver, and reducing cytotoxicity related to non-alcoholic fatty liver and JNK phosphorylation and inhibitory factors related to inflammation. Showed improvement effect. Therefore, the fermented ginseng according to the present invention can be used in health functional foods for preventing or improving non-alcoholic steatohepatitis.

1 is a view showing the measurement result of ginsenoside of fermented ginseng.
2 is a diagram showing the effect of improving non-alcoholic steatohepatitis in vivo of a fermented ginseng administration group.
3 is a diagram showing the effect of improving inflammation in the liver and reducing the expression of fibrosis genes in a fermented ginseng administration group.
4 and 5 are diagrams showing the effect of reducing lipid accumulation in the liver of the fermented ginseng administration group.
Figure 6 is a diagram showing the reduction of liver lipid peroxidation and JNK phosphorylation in the fermented ginseng administration group.
7 is a diagram showing the cytotoxicity and inhibition of inflammation related to non-alcoholic fatty liver in a fermented ginseng administration group.
8 is a diagram showing the inhibition of inflammation by fermented ginseng.

Terms or words used in this specification and claims should not be construed as being limited to their usual or dictionary meanings, and that the inventor may appropriately define the concept of terms in order to describe his own invention in the best way. Based on the principle, it should be interpreted as a meaning and concept consistent with the technical idea of the present invention.

Therefore, the matters described in the embodiments, reference examples, and drawings described in the present specification are only the most preferred examples of the present invention and do not represent all the technical ideas of the present invention, so that they can be replaced at the time of application. It should be understood that there may be various equivalents and variations.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are for illustrative purposes only, the scope of the present invention is not construed as being limited by these examples.

The present invention provides a health functional food composition for preventing or improving non-alcoholic steatohepatitis containing fermented ginseng.

In this case, the composition may be a highly concentrated solution, a powder, or a granule form, and the form of the food additive includes any one of powder, granule, tablet, capsule, and liquid.

In addition, the health functional composition of the present invention, health functional food or food additive comprising the same, in addition to including the extracts listed above, may further contain sweetening agents, flavoring agents, physiologically active ingredients, and minerals.

Preferably, the health functional foods are beverages, chips, noodles, processed livestock foods, processed seafood, processed dairy foods, fermented milk foods, beans foods, grain foods, microbial fermented foods, baked goods, seasonings, processed meats, acidic beverages, licorice It may be any one or more than one of a variety, herbs, more preferably a functional beverage, and the composition is preferably 1 to 50 parts by weight relative to the weight.

Example 1. Preparation of fermented ginseng (GBCK25)

Ginseng is a General Bio Co., Ltd. It was purchased from (Jeollabukdo, General Bio Co., Korea) and fermented ginseng using a standardized procedure. The ginseng fermentation process was carried out using the following new microbial strains. Saccharomyces servazzii GB-07 and pectinase enzyme were administered for 5 days.

That is, by complex fermentation with the new fungus Saccharomyces servazzii GB-07 and pectinase enzyme for 5 days, ginsenoside was converted into compound K, which is easily absorbed in vivo.

1) The first step of selecting and washing ginseng;

The ginseng used for the ginseng fermentation is Korean ginseng (Panax ginseng CA Meyer), Hwagi ginseng (Panax quinquefolium), Jeonchil ginseng (Panax notoginsneng), bamboo japonicum, Panax trifolium, and Himalayan ginseng (Panax pseudoginseng), Including one or more of Vietnamese ginseng (Panax vietnamensis), but wild ginseng, fresh ginseng, red ginseng, white ginseng, fine ginseng, ginseng leaves, ginseng fruit, red ginseng extract, wild ginseng stem cell, wild ginseng callus, ginseng stem cell, ginseng callus, wild ginseng cultured root, adventitious root , Camphor ginseng may contain one or more. Preferably, ginseng is washed by selecting 4-6 year old fresh ginseng.

2) a second step of cutting the washed ginseng into 01~1cm;

This is a step of cutting the washed ginseng into 01~1cm. When fermentation is performed by inoculating the enzyme strain mixture through the cutting, the contact area increases, thereby enhancing fermentation efficiency.

3) Mixing purified water with the cut ginseng, and inoculating a mixture of enzyme strains in which a pectinase enzyme and a Saccharomyces servazzii strain are mixed at a weight ratio of 2:3 at 30 to 60°C. A third step of fermenting for 5 days to prepare a fermented ginseng broth;

A reaction solution is prepared by mixing purified water with the cut ginseng. In addition, fermentation including a culture step of inoculating and culturing an enzyme selected from the pectinase family and an enzyme strain mixture comprising Saccharomyces servazzii strain GB-07 Ginseng is manufactured.

The enzymes of the pectinase family are Cytolase PCL5, Pyr-Flo, Rapidase C80Max, Optivin Mash, and Smizyme AC One selected from the group consisting of (Sumyzyme AC) is used as an enzyme.

Preferably, Cytolase PCL5, Pyr-Flo, Rapidase C80Max, Optivin Mash, and Sumyzyme AC. One selected from the group consisting of enzymes and Saccharomyces servazzii (Saccharomyces servazzii) strain GB-07 are mixed in a ratio of 1 to 3 to 1 to 3 to prepare an enzyme strain.

Purified water is mixed with the cut ginseng, and the enzyme strain mixture is inoculated and fermented for 5 days at 30 to 60°C. 10-20 parts by weight of the enzyme strain mixture is inoculated and fermented.

4) The fourth step of preparing fermented ginseng by concentrating, drying and pulverizing the ginseng fermentation broth

The method of concentrating the ginseng fermentation broth is a step of concentrating the extract by vacuum concentration and low temperature vacuum concentration. In the drying method, low-temperature vacuum drying is a method of drying by maintaining the pressure inside the dryer in a vacuum and adjusting the temperature to about 5~15℃. There is no denaturation of the extracted components, and the taste and aroma are not lost. Also, cold air drying, hot air Drying, freeze drying, far-infrared drying, and vacuum drying may be used.

The pulverization method is a step of pulverizing to a certain size to complete ginseng powder, and the dried ginseng fermentation broth is pulverized using a cutter mill, jet mill, and hi jet mill. The ginseng fermented powder is completed.

Experimental Example 1. Ginsenoside analysis of fermented ginseng

Fermented ginseng was dissolved in a modified Krebs-Henseleit (KH) buffer consisting of 120.0mM NaCl, 1.2mM MgSO4, 4.8mM KCl, 1.2mM KH2PO4, 25mM NaHCO3, 11.0mM glucose and 25mM CaCl2.

For HPLC analysis, a YL 9100 HPLC system was used, and a YMC-Triart C18 column (250 mm long, 46 mm inner diameter) was used. The detection wavelength of the detector was UV 230 nm, the column temperature was analyzed at 35°C, and the mobile phase was aceto. It was analyzed at a flow rate of 08 mL/min using nitrile and water. All reagents for the experiment were purchased and used from Sigma Aldrich (St. Louis, MO, USA).

1 is a view showing the measurement result of ginsenoside of fermented ginseng.

As a result of measuring ginsenosides in fermented ginseng, the content of ginsenosides in the fermented ginseng was ginsenoside-Rg1 18.71 mg/g, Re 43.51 mg/g, Rh1(s)+ Rg2(s) 40.45 mg/g, Rb2 18.73 mg/g, Rd 10.23 mg/g, Rg3(s) 4.36 mg/g, CK 23.32 mg/g and small amounts of other ginsenosides were measured.

Experimental Example 2. Evaluation of efficacy for improving non-alcoholic steatohepatitis

1-1. Experiment method

1) Experimental animals

Male 6-week-old C57BL/6 wild-type mice were purchased from Samtako Bio Korea (O-San, Korea) and used in this study. Rats were placed under standard conditions (24 ± 3 ℃, humidity 50 ~ 5%) and ingested a sterile normal diet (ND) and water. In accordance with the "Institutional Animal Care and Use Committee Guidelines" of Chonbuk National University (Jeonju, Korea), the laboratory animal care principles were followed.

To induce, they were fed a normal diet (ND) or WD diet (WD) for 12 weeks. The WD diet is a dietary control consisting of high in fat (40 kcal%), fructose (20 kcal%), and 2% cholesterol.

The experimental group was orally administered fermented ginseng at a dose of 10 mg/kg, 20 mg/kg, 100 mg/kg, 200 mg/kg, or 400 mg/kg once a day.

Fermented ginseng was dissolved in distilled water through sonication and orally administered once a day for 12 weeks in each WD diet group.

2) histopathological analysis

After fixing the tissue with 10% neutral buffered formalin solution, the liver tissue was finally inserted into paraffin using standard protocols.

Tissue sections (6 mm) were prepared using an HM-340E microtome (Thermo Fisher Scientific Inc., Waltham, MA, USA) and stained with hematoxylin and eosin (H&E) according to standard methods.

Histopathological analysis of liver sections was performed using an optical microscope (BX-51 microscope; Olympus Corp., Tokyo, Japan).

3) Non-alcoholic fatty liver disease (NAFLD)

To confirm the severity of NASH (Nonalcoholic steatohepatitis), liver sections stained with H & E were evaluated with the NAFLD activity score (NAS) system.

The system considered the degree of steatosis (0-3), inflammation of the lobules (0-3), and degeneration of hepatocytes (0-2). The NAS, the sum of these scores, represents the severity of NAFLD and/or NASH.

4) Oil Red O staining

Oil Red O staining kit (Scytek Lab., Logan, UT, USA) was used to measure lipid accumulation in hepatocytes.

Briefly, liver sections were washed with propylene glycol for 5 minutes at room temperature, stained with a heated (60 ℃) Oil Red O solution for 20 minutes, and counter-stained with hematoxylin for 1 minute. Oil Red O staining analysis and digital imaging software (analySIS TS, Olympus Corp.) were performed using an optical microscope.

5) Measurement of serum and liver biochemistry

Liver damage was assessed by measuring serum concentrations of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) with the AM101-K spectrophotometric assay kit (ASAN Pharmaceutical, Hwasung, Korea).

Liver fat accumulation was measured by measuring triglyceride (TG) and total cholesterol (TC) content with an AM202-K spectrophotometer assay kit (ASAN Pharmaceutical). The absorbance of the sample was quantified at a wavelength of 490 nm or 550 nm using an EMax spectrophotometer (Molecular Devices, Sunnyvale, CA, USA).

6) Quantitative real-time polymerase chain reaction

RNA was extracted from liver tissue using a hybrid-R RNA purification kit (GeneAll Biotechnology Co., Ltd., Seoul, Korea).

After incubation with DNase I containing an RNase inhibitor (TOYOBO, Osaka, Japan), samples were transcribed using ReverTra Ace qPCR RT Master Mix (TOYOBO) according to the manufacturer's instructions. The specificity was confirmed by melting curve analysis after the quantitative real-time polymerase chain reaction (qRT-PCR) reaction was completed in the CFX96 Real-Time PCR Detection System (Bio-Rad Laboratories, Hercules, CA, USA) using SYBR Green. . Relative quantification was performed by normalizing to the value of ribosomal protein lateral stalk subunit P0 (RPLP0).

7) Cell culture and Kupffer cell separation

The alpha mouse liver 12 (AML12) cell line was provided by Professor Heung-Sik Choi of Chonnam National University.

AML12 cells were cultured with DMEM containing 10% FBS, Insuline Transferrine Seleniume Pyruvate, 40 ng/mL, 100 IU/mL penicillin and 100 mg/mL streptomycin, and cultured at 37°C in a 5% CO ₂ incubator. For in vitro experiments, cells were cultured for 24 h in serum-free medium.

After incubation, the cells were stimulated with palmitic acid or fermented ginseng (1mg/mL, 2mg/mL and 4mg/mL) treatment for 24 hours.

Murine monocyte/macrophage cell line RAW264.7 was purchased from the American Type Culture Collection (Rockville, MD, USA), and DMEM (WELGENE Inc.) containing 10% FBS, 100IU/mL penicillin was used in a 5% CO ₂ incubator at 37°C. Cultured.

Kupffer cells (KCs) were isolated. Briefly, mouse livers are separated into two layers by centrifugation with 70% and 40% Percoll. Cells were collected at the interface and KC was actively selected using a magnetic cell selection method using an anti-F4/80 antibody. Inflammation was induced by treatment with 1 mg/mL LPS, and fermented ginseng was treated for 24 hours.

8) Cell viability analysis

Cells were seeded in 96-well plates (1×10⁴ cells/well) and attached to serum-free medium for 24 hours. After 24 hours, the fermented ginseng was treated with the cells at 0-400 μg/ml for 24 hours, and then cell viability was measured using CellTiter 96. The absorbance was quantified at 490 nm using the spectrophotometer described above.

9) Intracellular TG measurement

To measure the level of TG in cells, cells were inoculated into 96-wells, and then fermented ginseng was cultured at 0-400µg/ml for 24 hours. After incubation for 24 hours, the medium was removed and the cells were washed several times with phosphate buffered saline. After completely removing phosphate buffered saline, 10 mL dimethylsulfoxide was added for 1 minute to extract intracellular TG from each well. The absorbance of the sample was measured at a wavelength of 550 nm.

10) Malondialdehyde assay

To evaluate lipid peroxidation in liver tissue, thiobarbituric acid (TBARS) in the form of malondialdehyde (MDA) was measured using the oxalSelect TBARS Assay Kit.

11) Western blot assay

Liver tissues or cells were lysed directly on ice for 30 min with extraction buffer. Protein concentration was measured using the Pierce BCA Protein Assay Kit according to the manufacturer's protocol.

After dodecyl sulfateepolyacrylamide gel electrophoresis, the gel was transferred to a polyvinylidene difluoride membrane and then blocked with 5% FBS in Tris-buffered saline containing 0.05% Tween-20 for 1 hour at room temperature.

The primary antibody was diluted 1:1,000 with a blocking solution and incubated at 4°C overnight. Antibodies specific to CYP2E1 (Abcam, Cambridge, UK), c-Jun N-terminal kinase (JNK), phospho JNK (pJNK) and b-actin were used.

Antigen-antibody complexes of polyvinylidene difluoride membranes were detected using a secondary horseradish peroxidase-conjugated IgG antibody.

Immunoblot images were visualized with ImageQuant LAS 500. ImageQuant TL software was used to quantify the expression level of the protein bands.

12) enzyme immunoassay

To measure hepatocyte inflammatory cytokines, proteins were extracted. Protein levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and IL-1 beta (IL-1b) were measured using an ELISA kit (e-Bioscience, San Diego, CA, USA) I did. The absorbance at 450 nm was measured.

1-2. Experiment result

1) Efficacy in improving non-alcoholic steatohepatitis in fermented ginseng administration group

2 is a diagram showing the effect of improving non-alcoholic steatohepatitis in vivo of a fermented ginseng administration group.

In Figure 2, (A) H&E stained liver section, (B) body weight and liver weight change, (C) NAFLD-related factor measurement, (D) liver damage and lipid accumulation-related factors due to non-alcoholic steatohepatitis were measured. Indicated.

As a result of administration of fermented ginseng in WD-fed mice, it was confirmed that fat accumulation and inflammation in hepatocytes were reduced.

In addition, WD-fed mice showed an increased body weight compared to the control group, but there was no significant difference due to a decrease in body weight due to administration of fermented ginseng. Accordingly, as a result of measuring liver weight, it was increased in WD-fed mice compared to the control group, and as a result of administration of fermented ginseng to WD-fed mice, liver weight was generally decreased.

The NAS associated with the degree of steatosis, inflammation and febrile degeneration associated with non-alcoholic steatohepatitis was significantly decreased in the WD-administered mice compared to the WD-administered mice regardless of the fermented ginseng concentration.

In addition, as a result of measuring the enzyme activity in fatty liver, ALT was significantly decreased by administration of fermented ginseng.

As a result of histopathological analysis, liver TC and TG decreased with the dose of fermented ginseng.

Therefore, this result can be used as a health functional food composition for preventing or improving non-alcoholic steatohepatitis because liver damage caused by non-alcoholic steatohepatitis is significantly reduced by ingestion of fermented ginseng.

2) Non-alcoholic steatohepatitis-related inflammation and liver fibrosis improvement effect

3 is a diagram showing the effect of improving inflammation in the liver and reducing the expression of fibrosis genes in a fermented ginseng administration group.

To investigate the effect of fermented ginseng on liver inflammation and liver fibrosis caused by non-alcoholic steatohepatitis, qRT-PCR was used to measure mRNA expression of related liver genes.

The mRNA expression of TNF-α was significantly decreased in the liver tissues of WD-fed mice administered with fermented ginseng.

Liver mRNA levels of IL-6 were significantly reduced in WD-fed mice administered all concentrations of GBCK25. These were not observed at the liver mRNA level of IL-1β.

However, these inflammatory cytokines were significantly reduced in liver tissues of WD-fed mice administered fermented ginseng.

Fermented ginseng administration reduced inflammation due to non-alcoholic steatohepatitis, and decreased mRNA expression of α-SMA, metalloproteinase-1 (TIMP-1), and Col-1, which are related to liver fibrosis.

Therefore, it can be confirmed that the WD diet mice are improved in relation to inflammation and fibrosis caused by non-alcoholic steatohepatitis.

3) Efficacy in reducing hepatocyte lipid accumulation

4 and 5 are diagrams showing the effect of reducing lipid accumulation in the liver of the fermented ginseng administration group.

In order to evaluate whether the administration of fermented ginseng affects hepatic steatosis, it was measured by staining with Oil Red O in the liver tissue.

As a result, compared to the control group, intrahepatic steatosis was confirmed in the WD-fed mice, and the fat was decreased in a concentration-dependent manner in the fermented ginseng administration group.

Accordingly, mRNA expression of fatty acid synthase (FAS), acetyl CoA carboxylase (ACCa), and diacyl glycerol O-acyl transferase 1 (DGAT1) in liver liposynthetic genes was measured.

ACCa was significantly decreased in the fermented ginseng-administered group, and intrahepatic TG was stored or released along with VLDL (very-low-density lipoprotein) particles, such as microsomal TG transfer protein (MTP) and apolipoprotein B (ApoB). Genes related to VLDL synthesis were measured.

In addition, fatty acid oxidation-related genes carnitine palmitoyltransferase 1 (CPT1), CYP2E1, CYP3A11, and CYP4A10 were measured.

As a result, the expression level of CYP2E1 was significantly decreased in a dose-dependent manner in the fermented ginseng administration group.

In particular, it can be confirmed that the CYP2E1 protein was decreased in the liver of mice administered with fermented ginseng.

Taken together, these results are thought to be able to reduce lipid accumulation caused by non-alcoholic steatohepatitis by ingesting fermented ginseng.

4) Efficacy in reducing hepatocyte oxidative stress and JNK phosphorylation

Figure 6 is a diagram showing the reduction of liver lipid peroxidation and JNK phosphorylation in the fermented ginseng administration group.

Since non-alcoholic steatohepatitis may cause lipid peroxidation in hepatocytes or damage to adipocytes, MDA was measured to evaluate the level of lipid peroxidation.

As a result, it was confirmed that liver MDA was inhibited in the fermented ginseng administration group, which is thought to be inhibited by fermented ginseng from lipid peroxidation and damage to fat cells in hepatocytes caused by steatohepatitis.

To confirm the effect of fermented ginseng on hepatocellular damage caused by lipid peroxidation, the effect on JNK phosphorylation in hepatocellular lipoproteinosis induced by free fatty acid (FFA) was analyzed.

JNK was phosphorylated in the WD diet group, a non-alcoholic steatohepatitis inducing group, but decreased in a concentration-dependent manner by administration of fermented ginseng. Reduced CYP2E1 mediated lipid peroxidation is associated with the cellular JNK pathway leading to cellular damage.

Therefore, it is considered that fermented ginseng has protective activity against fat toxicity caused by non-alcoholic steatohepatitis in mice.

5) Efficacy in inhibiting oxidation and liver inflammation in fatty liver cells

7 is a diagram showing the cytotoxicity and inhibition of inflammation related to non-alcoholic fatty liver in a fermented ginseng administration group.

8 is a diagram showing the inhibition of inflammation by fermented ginseng.

To clearly demonstrate the effect of fermented ginseng on non-alcoholic fatty liver-related hepatocyte damage and inflammation, experiments were performed at the level of AL12 or RAW 264.7 cells according to PA or LPS stimulation.

After measuring the concentration of fermented ginseng without intracellular toxicity in AML12 cells, the effects of fermented ginseng on cell damage and lipid accumulation in PA-treated AML12 cells were evaluated.

As a result, in the fermented ginseng treatment group, the intracellular TG and lipid-generating FAS and the mRNA expression levels of ACCa were significantly reduced.

This effect was also observed at the mRNA expression level of CYP2E1, indicating that hepatic steatosis and CYP2E1-mediated oxidative stress in fermented ginseng were reduced, resulting in a reduction in the severity of non-alcoholic steatohepatitis.

As a result of Western blot analysis, protein levels of CYP2E1 and pJNK were decreased in PA-treated AML12 cells by fermented ginseng treatment.

After measuring the concentration of fermented ginseng that did not show cytotoxicity in RAW264.7 cells and primary KC, the mRNA expression level of pre-inflammatory genes was significantly reduced by treatment with fermented ginseng in RAW264.7 cells treated with LPS and primary KC. .

As a result of the above experiment, fermented ginseng improves the severity of non-alcoholic steatitis by affecting lipid accumulation, oxidative stress-mediated cell damage and inflammation.

Claims (1)

A first step of selecting and washing ginseng;
A second step of cutting the washed ginseng into 0.1 to 1 cm;
Purified water was mixed with the cut ginseng, and the enzyme strain mixture was inoculated with a pectinase enzyme and a 2:3 mixture of Saccharomyces servazzii strain based on the weight ratio and inoculated at 30-60° C. for 5 days. A third step of fermenting to prepare a ginseng fermented liquid; And
Prevention or improvement of non-alcoholic steatohepatitis containing fermented ginseng as an active ingredient by concentrating the ginseng fermentation broth, followed by low-temperature vacuum drying at a temperature of 5 to 15°C, and pulverizing to prepare fermented ginseng In the health functional food composition for,
The composition is a health functional food composition for preventing or improving non-alcoholic steatohepatitis, characterized in that by orally administered at a concentration of 200 mg/kg based on C57BL/6 mice to reduce the expression of non-alcoholic steatohepatitis-related inflammatory genes.

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