KR20150069174A - Composition for preventing or treating non-alcoholic fatty liver containing Nontoxic fermented extract of Rhus verniciflua as an active ingredient - Google Patents
Composition for preventing or treating non-alcoholic fatty liver containing Nontoxic fermented extract of Rhus verniciflua as an active ingredient Download PDFInfo
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- KR20150069174A KR20150069174A KR1020130155276A KR20130155276A KR20150069174A KR 20150069174 A KR20150069174 A KR 20150069174A KR 1020130155276 A KR1020130155276 A KR 1020130155276A KR 20130155276 A KR20130155276 A KR 20130155276A KR 20150069174 A KR20150069174 A KR 20150069174A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/22—Anacardiaceae (Sumac family), e.g. smoketree, sumac or poison oak
Abstract
Description
The present invention relates to a composition for preventing or treating nonalcoholic fatty liver comprising detoxified extract of Rhus verniciflua extract as an active ingredient, and more particularly, to a composition for preventing or treating nonalcoholic fatty liver, Inoculation of Saccharomyces carlsbergensis strain and cultivation to remove the toxic components of the lacquer shell, as well as the nonalcoholic fatty liver prevention Or therapeutic compositions.
Rhus Verniciflua Stokes ( Rhus Verniciflua Stokes) is a deciduous broad-leaved arboreous tree belonging to the Anacardiaceae family, which is known as the origin of the Tibetan and Himalayan regions of Central Asia. It is 20 m high and 60 cm in diameter. Is a kind of secretion of plant physiological phase, which is collected in a short time. It is an off-white oily liquid with a sweet and bitter taste. When it comes in contact with air, it turns brown. Urushiol, which is the main component of chalk, has been widely used as a chill, since it is hardened by enzyme reaction when it comes in contact with oxygen in the air, and forms a fine coating, which is a polymer of three dimensional structure unlike other paints.
There are about 70 species of lacquer plants in Korea and about 600 species in the world. Six species such as lacquer tree, lacquer tree, vine lacquer tree, rush tree, lacquer tree, and lacquer tree are grown in Korea.
Lacquer is a mysterious medicinal substance which has both toxicity and weakness. In the Orient, the lacquer has been used for edible and medicinal purposes from the old days, and the prescriptions which use it for folk remedies such as ejaculation, insect repellent, Is also introduced as a medication to treat gastrointestinal diseases, constipation, and ejaculation.
Recently, studies have shown that Urushiol, which is the main ingredient of rasse lacquer and has allergies, has strong anticancer, antioxidant and antimicrobial activity, and the flavonoid component extracted from the shell of lacquer tree and woody part of the lacquer, Formation inhibitory action, inhibiting the proliferation and metastasis of cancer cells, inducing the differentiation of cancer cells into normal cells, and also showed antioxidant, hangover and gastritis inhibitory effects.
When the poisonous substances such as urushiol directly cause ingestion of poisonous lacquer, which has various kinds of pharmacological effects, such as rash, rash, and itching, side effects appear.
In addition, according to the method of putting together the lacquer with the jujube, chestnut, oranges in the chicken or duck, it is found that one of the three people who ingest the lacquer chicken or the lacquer duck has various side effects such as dermatitis, A method is urgently required.
Korean Patent Registration No. 10-0367286 (Oct. 10, 2003) discloses a method for extracting toxic russus extracts comprising a step of hot air drying, a step of adding a solvent and heat treatment, and a concentration step of concentrating the raspberry extract And Korean Patent Publication No. 10-0339837 (Jun. 2002) discloses a method of drying at 170 캜 for 4 hours or more in order to remove the lacquer poison from the lacquer tree.
The heat treatment method is disadvantageous in that it is inconvenient in the manufacturing process because it requires heat treatment process at high temperature and high pressure and other useful pharmacological components can be destroyed under high temperature and high pressure. Fermentation methods are proposed to overcome these disadvantages.
For example, in the Korean Patent Registration No. 10-0512328 (2005.09.05.), The lacquer tree is grown and inoculated with Aspergillus oryzae, Aspergillus kawachii, A method for producing a lacquer with toxicity removed is disclosed.
This method uses stalks and branches of Rhus verniciflua separately. Stalk and branches of Rhus verniciflua are weakly toxic, which is easy to detoxify, but has a disadvantage that it contains a small amount of medicinal ingredients.
On the other hand, the lacquer shell has the most medicinal ingredient, but it has strong toxicity. Therefore, it is necessary to study the method of detoxifying the liquorice shell containing the most medicinal ingredient.
The inventors of the present invention have studied the method of detoxifying the lacquer shell and found that the toxic component of Rhus verniciflua can be removed by inoculating and culturing the Saccharomyces carlsbergensis strain in the extract of the lacquer tree and the woody part of the lacquer tree And a method of detoxifying the extract of Rhus verniciflua and a detoxified extract of Rhus verniciflua produced by using the extract. (Korean Patent Publication No. 10-1089176, Dec. 2, 2011 .)
Korean Patent Laid-Open Publication No. 10-2013-0040663 (Apr. 24, 2013) discloses a composition for improving and preventing hyperlipidemia and fatty liver, which contains Rhus verniciflua extract. The Rhus verniciflua extract uses Rhus verniciflua var. The wood part has a weak toxicity but has a disadvantage that it contains a small amount of medicinal component.
To date, there have been no studies on the efficacy of non-alcoholic fatty liver disease treatment of extracts of toxins that have been removed from poisons.
The object of the present invention is to remove the toxic components of the lacquer tree by inoculating and culturing the Saccharomyces carlsbergensis strain in the extract of Rhus verniciflua obtained by hydrolysis of the lacquer fermented with the enzyme, The present invention provides a composition for preventing or treating nonalcoholic fatty liver which contains as an active ingredient a detoxified extract of Rhus verniciflua extract capable of inhibiting the accumulation.
In order to achieve the above object, the present invention provides the following means.
The present invention relates to a method for producing an extract of Rhus verniciflua, which is obtained by hot-extracting Rhus verniciflua fermented with an enzyme, and inoculating Saccharomyces carlsbergensis strain with the lactase, amylase, protease Lipase, Sucrase, Maltase, Cellulase, Bromelain, or a mixture thereof. The culture is incubated at 25-35 ° C for 48-72 hours The present invention provides a composition for prevention or treatment of nonalcoholic fatty liver comprising as an active ingredient a detoxified ricinic fermented product extracted from rhizome.
The composition can reduce liver fat.
The composition may increase the expression of phosphorylated AMPK (p-AMPK) or PPAR alpha (Peroxisome proliferator activated receptor alpha).
The composition may reduce the expression of FAS (Fatty acid synthease) or SREBP-1 (sterol-regulated-element binding protein-1).
The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of the composition; And a pharmaceutically acceptable carrier. The present invention also provides a pharmaceutical composition for prevention or treatment of fatty liver.
The present invention also relates to a food-effective amount of the composition; And a food-acceptable carrier.
The composition for prevention or treatment of nonalcoholic fatty liver comprising the detoxified extract of Rhus verniciflua according to the present invention as an active ingredient is characterized in that the extract is a mixture of Saccharomyces carlsbergensis strain To remove toxic components of the lacquer shell and to inhibit fat accumulation in nonalcoholic fatty liver patients.
FIG. 1 is a graph showing the effect of the detoxified Rhus verniciflua extract fermented according to the present invention on cell growth upon treatment at various concentrations.
FIG. 2 is a graph showing the effect of inducing fat as an oleic acid in HepG2 cells and inhibiting fat induction upon treatment of the detoxified rhus verniciflua extract according to the present invention at various concentrations.
FIG. 3 is a graph showing the inhibition mechanism of fat induction by Western blot when the fat-induced lipid was induced into oleic acid in HepG2 cells and then treated with the detoxified rhus verniciflua extract according to the present invention at various concentrations.
Hereinafter, the present invention will be described in detail.
Non-alcoholic fatty liver disease (NAFLD) is a disease in which hepatic triglycerides accumulate in the liver regardless of alcohol, including steatosis and non-alcoholic steatohepatitis (NASH) ). NASH is characterized by inflammation or fibrosis with fatty liver. On the other hand, simple fatty liver is thought to be a benign disease with good clinical prognosis, but NASH is a progressive liver disease and is recognized as a precursor disease causing cirrhosis or liver cancer. The risk factors for nonalcoholic fatty liver disease are largely obesity, diabetes, and hyperlipemia.
The causes of nonalcoholic fatty liver disease can be explained by two mechanisms. The first step is to become a healthy liver fatty liver, resulting from the increase of fatty acid inflow from adipose tissue to liver due to insulin resistance at the periphery, and the second step is the peroxidation of fat, infiltration of inflammatory cells, Is the process by which oxygen free radicals and FAS ligands are activated. Thus, the increase in fatty liver increases the expression of CYP2E1 (cytochrome peroxidase 2E1), induces lipid peroxidation in the liver cell membrane by producing reactive oxygen species, and increases in LPS and oxidative stress increase TNF- Induce apoptosis and promote liver damage.
There are two broad categories of therapeutic agents used in patients with nonalcoholic fatty liver disease. The first is a drug that treats and improves fatty liver through the correction of risk factors such as orlistat, insulin resistance treatment (metformin, pioglitazone, rosiglitazone) or drugs for treatment of hyperlipemia (clofibrate, gemfibrozil, bezafibrate, atorvastatin, simvastatin) The second is a drug that regenerates hepatocyte damage independently of risk factors for nonalcoholic fatty liver disease, including ursodeoxycholic acid and taurine, antioxidant (vitamin E), and nutritional supplements (lecithin, betaine, N -acetylcysteine).
Recently, there has been a growing interest in extracts of natural materials that are effective in the treatment of patients with nonalcoholic fatty liver without side effects.
The applicant of the present invention has studied how to detoxify the lacquer shell and found that the toxic components of Rhus verniciflua can be removed by inoculating and cultivating Saccharomyces carlsbergensis strain in the extract of lacquer tree and woody part The method of detoxifying the extract of Rhus verniciflua and the detoxified extract of Rhus verniciflua produced by using it was filed and registered. (Korean Registered Patent No. 10-1089176, December 22, 2011)
The Applicant has confirmed that the registered detoxified Rhus verniciflua extract fermented product can be used as an active ingredient of a composition for the prevention and treatment of a non-alcoholic fatty liver model, thereby completing the present invention.
First, a composition for preventing or treating nonalcoholic fatty liver comprising the detoxified rhus verniciflua extract as an active ingredient according to the present invention will be described.
The lacquer used in the composition of the present invention may be a mixture of lacquer shell and woody part. The mixing ratio of the shell to the woody part may be 1: 0.1 to 0.1: 1 by weight, but is not limited thereto. It is preferable that the shell and the woody part of the Rhus verniciflua are pulverized and used.
The detoxified ricin-extracted fermented product of the present invention can be produced by inoculating and cultivating Saccharomyces carlsbergensis strain into Rhus verniciflua extract obtained by hot extraction of Rhus verniciflua fermented with an enzyme.
The fermentation of Rhus verniciflua by enzymatic hydrothermal extraction has the advantage that various components of Rhus verniciflua can be extracted more by enzymatic degradation.
In the present invention, the enzyme may be a lactase, an amylase, a protease, a lipase, a sucrase, a maltase, a cellulase, a bromelain Bromelain, or mixtures thereof.
In one embodiment of the invention, the enzyme is a multi-atom ® (trade name, 8 species complex enzymes: papain (PAPAIN) 3,450 NF, amylase (AMYLASE) 2,500 DU, protease (PROTEASE) 10,000 HUT, malteu diahseutaahje (MALT DIASTASE) 270 DP,
In another embodiment of the present invention, the enzyme may be SpezymePrim (product name, containing 75% of alpha amylase, manufacturer: Genencor International, Belgium).
In the present invention, the amount of the enzyme to be used is preferably 0.01 to 0.2% by weight based on the substrate, and is not limited thereto.
The Rhus verniciflua extract can be obtained by subjecting Rhus verniciflua fermented with the enzyme to hot water extraction. The hot water extraction can be carried out at a temperature of 80 to 130 ° C for 6 to 8 hours and can be carried out under a pressure of 2 atm or more, but is not limited thereto. The water may be ordinary water, preferably distilled water, deep sea water, or a mixture thereof, but is not limited thereto. At this time, the mixing ratio of the distilled water to the deep sea water may be preferably 5: 1 to 1: 5, more preferably 1: 1, but is not limited thereto.
In the present invention, the strain is preferably a strain of Saccharomyces carlsbergensis . It is preferable that the strain is added in an amount of 0.1 to 0.3 part by weight based on 100 parts by weight of the Rhus verniciflua extract. If the added amount is less than 0.1 part by weight, the fermentation rate is slowed. If the added amount is more than 0.3 part by weight, Therefore, there is an uneconomical problem.
The culture is preferably carried out at 25 to 35 ° C. for 48 to 72 hours. If the temperature is lower than 25 ° C., the fermentation efficiency is lowered. If the temperature is higher than 35 ° C., . When the incubation time is less than 48 hours, there is a problem that toxic components of Rhus verniciflua can not be sufficiently removed, and when the time exceeds 72 hours, there is a problem that the germs can reproduce.
In the present invention, the culture is performed in a closed fermentation tank, and purified air is preferably injected when the air is injected into the fermentation tank.
The detoxified Rhus verniciflua fermented product used in the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze drying or spray drying.
The composition of the present invention is very effective for preventing or treating nonalcoholic fatty liver.
As used herein, the term "fatty liver" refers to a state in which fat is accumulated in hepatocytes in an excessive amount due to liver fat metabolism disorder. It causes various diseases such as angina pectoris, myocardial infarction, stroke, atherosclerosis and pancreatitis.
The composition containing the detoxified rhus verniciflua extract of the present invention has very excellent activity for the improvement of non-alcoholic fatty liver among various diseases of metabolic diseases. The composition of the present invention can prevent or treat non-alcoholic fatty liver by various activities.
The composition of the present invention exhibits activity of preventing or treating fatty liver by significantly reducing liver fat.
The term "liver" as used herein includes cells or tissues.
The composition of the present invention increases the expression of phosphorylated AMPK (p-AMPK) or PPAR alpha (Peroxisome proliferator activated receptor alpha). AMP-activated protein kinase (AMPK) is an enzyme that is activated when intracellular energy is deficient and increases energy production. Phosphorylated AMPK (p-AMPK) inhibits the phosphorylation of ACC (acetyl-CoA carboxylase) and regulates the uptake of fatty acids into the mitochondria, inhibiting lipid synthesis.
In addition, the composition of the present invention reduces the expression of FAS (Fatty acid synthease) or SREBP-1 (sterol-regulated-element binding protein-1). SREBP-1 plays a major role in regulating genes involved in the synthesis of triglycerides in the liver.
The compositions of the present invention may be prepared with pharmaceutical compositions.
According to a preferred embodiment of the present invention, the composition of the present invention comprises a pharmaceutically effective amount of the detoxified rhus verniciflua extract of the present invention; And a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve efficacy or activity of the detoxified rhus vernicifluum extract described above.
When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier to be included in the pharmaceutical composition of the present invention is one commonly used in the present invention and includes lactose, textol, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components.
The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably administered orally.
The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Typical dosages of the pharmaceutical compositions of the invention are in the range of 0.4-180 mg / kg on an adult basis.
The pharmaceutical composition of the present invention may be prepared in a unit dose form by formulating it with a pharmaceutically acceptable carrier or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs Into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
The composition of the present invention can be provided as a food composition.
According to a preferred embodiment of the present invention, the composition of the present invention is a food-effective amount of the detoxified rhus verniciflua extract of the present invention described above; And a pharmaceutically acceptable carrier.
When the composition of the present invention is prepared as a food composition, it contains not only detoxified ricin-extracted fermented product as an active ingredient, but also ingredients normally added in the manufacture of food, for example, protein, carbohydrate, fat, Seasoning and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavors (saccharin, aspartame, etc.) may be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, etc., Can be added.
Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These embodiments are only for illustrating the present invention, and the scope of the present invention is not limited by these embodiments.
After collecting the lacquer tree, the shell of lacquer tree and the woody part were crushed. 20% by weight of the crushed stems of Rhus verniciflua and 80% by weight of woody parts were mixed. After the mixed lacquer pulverized with respect to 1,000 parts by weight to 1 part by weight was added to a multi-atom ® fermentation at 60 ℃ for 60 hours, the water content was air-dried to a 13 ~ 14%. 20 parts by weight of water (deep sea water: distilled water = 1: 1) was added to 1 part by weight of the lacquer tree fermented with the enzyme, and the mixture was extracted at 110 DEG C for 8 hours to obtain Rhus verniciflua extract.
100 parts by weight of the Rhus verniciflua extract was added to a 500 ml Erlenmeyer flask and 0.2 part by weight of Saccharomyces carlsbergensis strain was added thereto. The mixture was inoculated at 30 ° C for 72 hours at 150 rpm for detoxification. Water was prepared.
[Comparative Example 1]
Atorvastatin (AT), an antihyperlipidemic agent for lowering triglyceride, cholesterol, was prepared.
[Experimental Example 1]
HepG2, a hepatocellular cell line, was dispensed into 96-well plates at a density of 1 × 10 4, and cultured for 24 hours. Then, the detoxified rhus verniciflua extract (CO) prepared in Example 1 was treated for each concentration and reacted for 24 hours. The growth of the cell line using the (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyl-2H-tetrazolium bromide) reagent is shown in FIG.
The effect of the detoxified rhus verniciflua extract fermented in Example 1 on the abnormal hepatocyte HepG2 cell growth was confirmed to have no significant effect on cell growth even after treatment at a maximum concentration of 800 μg / ml.
[Experimental Example 2]
HepG2 was dispensed into 6-well plates at a density of 1 × 10 5, and cultured for 24 hours. Oleic acid was treated with 200 μM to induce fat, and the detoxified ricin-fermented extract prepared in Example 1 was treated at different concentrations And reacted for 24 hours. Thereafter, the cells were stained with oil-red-o (ORO) for 10 minutes, and then dissolved in isopropanol. The absorbance was measured using a microplate reader (Molecular Devices Co.)
Induced lipid accumulation in abnormal hepatocyte HepG2 and inhibition of fat induction of the detoxified extract of Rhus verniciflua extract prepared in Example 1. As a result, it was confirmed that even when 200 μg / ml was treated, fat induction was significantly inhibited, and 400 μg / Ml treatment.
In addition, when 3 μM of atorvastatin (AT), an anti-hyperlipidemic agent for lowering cholesterol, which is a triglyceride of Comparative Example 1, and 400 μg / ml of the detoxified rhus verniciflua fermented product prepared in Example 1 were treated Was confirmed.
[Experimental Example 3]
HepG2 was dispensed into 6-well plates at a density of 1 × 10 5, and cultured for 24 hours. Oleic acid was treated with 200 μM to induce fat, and the detoxified ricin-fermented extract prepared in Example 1 was treated at different concentrations And reacted for 24 hours. After that, western blotting was performed using SDS page gel using RIPA lysis buffer, and the result was confirmed with each antibody, as shown in FIG.
Induced lipid accumulation in hepatic hepatocyte HepG2 and lipid-induced inhibition of the detoxified extract of Rhus verniciflua extract prepared in Example 1 resulted in increased phosphorylated AMPK (p-AMPK) and PPARα (Peroxisome proliferator activated receptor alpha) It was confirmed that the increase of FAS (Fatty acid synthease) and SREBP-1 (sterol-regulated-element binding protein-1) was effectively reduced.
AMP-activated protein kinase (AMPK) is an enzyme that is activated when intracellular energy is deficient and increases energy production. Phosphorylated AMPK (p-AMPK) inhibits the phosphorylation of ACC (acetyl-CoA carboxylase) and regulates the uptake of fatty acids into the mitochondria, inhibiting lipid synthesis.
In particular, SREBP-1 plays a major role in regulating genes involved in the synthesis of triglycerides in the liver.
Therefore, the detoxified rhus vernicifera fermented product according to the present invention can be useful for treating non-alcoholic fatty liver by increasing phosphorylated AMPK (p-AMPK), decreasing SREBP-1 and decreasing liver fat .
Claims (6)
The enzyme may be selected from the group consisting of lactase, amylase, protease, lipase, sucrase, maltase, cellulase, Bromelain, Or a mixture thereof,
The composition for preventing or treating nonalcoholic fatty liver disease according to any one of claims 1 to 5, wherein the culture is performed at 25 to 35 DEG C for 48 to 72 hours.
A composition for prevention or treatment of nonalcoholic fatty liver disease comprising as an active ingredient a detoxified extract of Rhus verniciflua extract which reduces liver fat.
Wherein the composition comprises a detoxified extract of Rhus verniciflua extract which increases the expression of phosphorylated AMPK (p-AMPK) or PPAR alpha (Peroxisome proliferator activated receptor alpha) as an active ingredient.
The composition is a composition for preventing or treating nonalcoholic fatty liver comprising as an active ingredient a detoxified extract of Rhus verniciflua extract which reduces the expression of FAS (Fatty acid synthease) or SREBP-1 (sterol-regulated-element binding protein-1) .
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110062584A (en) * | 2016-10-07 | 2019-07-26 | 尹后男 | The preparation method of non-toxic paint solution and using its ferment rice bran powder and powder cereal nutriment preparation method |
| KR20220030449A (en) * | 2020-09-01 | 2022-03-11 | 농업회사법인 주식회사 옻가네 | Method for extracting collagen using rhus verniciflua fermentation broth and composition for extracting collagen containing nontoxic fermented extract of rhus verniciflua as an active ingredient |
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2013
- 2013-12-13 KR KR1020130155276A patent/KR20150069174A/en not_active Application Discontinuation
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110062584A (en) * | 2016-10-07 | 2019-07-26 | 尹后男 | The preparation method of non-toxic paint solution and using its ferment rice bran powder and powder cereal nutriment preparation method |
| KR20220030449A (en) * | 2020-09-01 | 2022-03-11 | 농업회사법인 주식회사 옻가네 | Method for extracting collagen using rhus verniciflua fermentation broth and composition for extracting collagen containing nontoxic fermented extract of rhus verniciflua as an active ingredient |
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