KR20120064218A - Anti-inflammatory effect of extracts of disporum sessile d.don - Google Patents
Anti-inflammatory effect of extracts of disporum sessile d.don Download PDFInfo
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- KR20120064218A KR20120064218A KR1020100125343A KR20100125343A KR20120064218A KR 20120064218 A KR20120064218 A KR 20120064218A KR 1020100125343 A KR1020100125343 A KR 1020100125343A KR 20100125343 A KR20100125343 A KR 20100125343A KR 20120064218 A KR20120064218 A KR 20120064218A
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- extract
- yunpanmul
- inflammatory
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Microbiology (AREA)
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- Polymers & Plastics (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a yunpanmul extract exhibiting anti-inflammatory activity. More specifically, the present invention relates to an anti-inflammatory composition containing the extract of Yunpanmul as an active ingredient by extracting the ground part of the Yunpanmul, the lily family, with methanol as a solvent. In addition, it was confirmed that the expression of iNOS (inducible Nitric Oxide Synthase), a gene that regulates the generation of nitric oxide, was also reduced. Therefore, Yunpan sprout extract has an effect that can be provided as an anti-inflammatory composition, processed foods, functional foods, or food additives that can suppress inflammation caused by inflammation-inducing substances.
Description
The present invention relates to a yunpanmul extract exhibiting anti-inflammatory activity. More specifically, the present invention relates to an anti-inflammatory composition containing the extract of Yunpannamul as an active ingredient by extracting the ground part of Yunpannaum, which is a lily plant, as a solvent, and an anti-inflammatory composition capable of suppressing inflammation caused by an inflammation-inducing substance. To provide a processed food, functional food, or food additives.
The present invention relates to a yunpanmul extract having an anti-inflammatory effect, in particular, to provide a food composition for health promotion using the yunpanmul extract.
An inflammatory response is a mechanism for repairing and repairing an injured area when an invasion that causes any organic change in the body or tissue, such as physical action, chemicals, or bacterial infection, is applied.
In other words, macrophages act when living organisms become infected with pathogens or when the immune system is hyperactive for some reason. Macrophages play an important role in defending the host against pathogens with the help of active agents of T-cells, which are pathogens and immune cells, and this activity is regulated by several mediators. In particular, the production of nitric oxide (NO) by nitric oxide synthase (NOS) of macrophages acts as an important mediator of killing or inhibiting growth of bacteria, fungi and parasites. .
Recent studies show that NO is a free radical produced from arginine by three major NOS isoforms: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). nNOS and eNOS are controlled by Ca 2 + / calmodulin (calmodulin), but, iNOS is regulated at the transcriptional level by inflammatory stimuli such as IL (interleukin), IFN (interferon), LPS. NO produced in a few seconds by nNOS or eNOS is responsible for normal physiological functions such as vasodilation, neurotransmission, and cellular destruction of pathogens, but overproduction of iNOS in macrophages reacts with superoxide. It forms peroxynitrite, which acts as a powerful oxidant, causing damage to cells and activating NFκB in macrophages activated by inflammatory stimuli, which are involved in chronic diseases such as inflammatory reactions, cancer, and atherosclerosis. Known. Korhonen. Et al., Current Drugs target ., 4 , pp 471-479, 2005; TJ GUZIK Et al., JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY ., 4, pp 469-487, 2003)
NO is known to be produced by iNOS in various pathophysiological processes, including inflammation and cancer. NOS is an important enzyme that regulates inflammation, vasodilation, neurotransmission, tumor cell and body homeostasis, and produces NO from L-arginine. In inflammatory conditions, macrophages produce large amounts of NO by iNOS. It is known to cause various chronic diseases. Expression of iNOS is induced by NFκB activity, which is an important mechanism by which LPS or cytokine is overproduced in macrophages.
Until now, there is no substance or product mainly composed of yunpanmul. In addition, since there are not many data showing anti-inflammatory activity among the efficacy of general health foods based on scientific basic data, it can be safely consumed using natural extracts, can suppress inflammatory reactions, and can be easily used in foods. There is a need for research on materials.
SUMMARY OF THE INVENTION The present invention is for the popular dissemination of yunpanmul, useful for preventing inflammatory diseases, and has an object of providing a processed beverage while exhibiting the pharmacological activity of the yunpanmul.
In addition, an object of the present invention is to provide a beverage composition, a food additive, etc. to which the Yunpannamul extract excellent in anti-inflammatory activity of the present invention is added.
Yunpanmul extract is washed with running water to remove foreign matter by washing with Yunpanmul (Disporum sessile D.Don), dried in sunny place, pulverized to powder, and deposited in 80% methanol, and the methanol deposit is ultrasonically After extracting three times for 30 minutes, the supernatant of the extract was recovered, concentrated under reduced pressure, the concentrate was suspended in water, the suspension was freeze-dried, and extracted with DMSO to prepare a yunpanmul extract. The NO production inhibitory activity and iNOS mRNA expression inhibitory effect of the Yunpannamul extract is confirmed to provide an extract that can be used as a food additive and a beverage composition of the Yunpannamul extract.
The present invention is a material having excellent effects such as acute inflammation suppression by the inflammatory response, the degree of expression of the inflammation-inducing substance and the amount of phosphorylated NFκB protein expression, and can produce a health functional food having an inflammatory disease alleviating activity by controlling the inflammatory response. have.
Also, because it is a plant, it can be used as a food additive and consumed. Accordingly, the present invention may exhibit an anti-inflammatory effect in addition to food, and may be used for inflammatory disease relief as a prophylactic agent.
Ingested by the method of the present invention, when ingesting a beverage composition for improving health comprising the Yunpannamul extract, will greatly help to improve the body's defense against cancer, bacteria, inflammation, diabetes and the like.
1 is a graph showing the results of inhibiting the production of NO in RAW 264.7 cells for Yunpanmul extract according to the present invention.
Figure 2 is a graph showing the RT-PCR results showing the inhibitory effect of iNOS mRNA expression in RAW 264.7 cells for Yunpanmul extract according to the present invention.
Figure 3 is a graph showing the RT-PCR results showing the inhibitory effect of COX-2 mRNA expression in RAW 264.7 cells for Yunpanmul extract according to the present invention.
4 is a graph showing the results of RT-PCR showing the inhibitory effect of TNF-α mRNA expression in RAW 264.7 cells against Yunpanmul extract according to the present invention.
5 is a graph showing the results of RT-PCR showing the inhibitory effect of IL-6 mRNA expression in RAW 264.7 cells for Yunpanmul extract according to the present invention.
Figure 6 is a Western blot results picture showing the inhibitory effect of iNOS expression at the protein level in RAW 264.7 cells for Yunpanmul extract according to the present invention.
7 is a photograph of Western blot showing the inhibitory effect of p-IκBα expression at the protein level in RAW 264.7 cells with respect to Yunpanmul extract according to the present invention.
8 is a photograph of Western blots showing the inhibitory effect of NFκB expression in RAW 264.7 cells moving into the nucleus in RAW 264.7 cells according to the present invention.
Figure 9 is a EMSA photograph showing the inhibitory effect of binding to the target DNA in the nucleus in RAW 264.7 cells with respect to Yunpanmul extract according to the present invention.
In this example, the following experiment was carried out to find out the anti-inflammatory activity in Yunpan sprout.
Example 1 Preparation of Yunpan Herb Extract
Stems and branches of the leafy leaf sprouts were dried and pulverized and extracted 300g of leafy leaf sprouts three times with 1.2 L of 80% methanol, and the obtained extract was concentrated using a rotary vacuum evaporator and then lyophilized to obtain 80% methanol extract.
Example 2 Measurement of Nitric Oxide (NO) Production
37 ° C., 5% carbon dioxide in DMEM medium containing 10% FBS, penicillin (100 unit / ml), streptomycin sulfate (100 μg / ml) in macrophage RAW 264.7 (1 × 10 4 cells / ml) on 96-well plate Incubated in the environment of, the extract (100, 50, 25, 12.5, 0 ㎍ / ㎖) was treated, and then treated with LPS (1 ㎍ / ㎖) was incubated for 24 hours. 100 μl of cultured cell medium was transferred to Griess reagent (5% (v / v), phosphoric acid and 0.1% (w / v) naphthylethylenediamine-hydrochloric acid in the same amount 1% (w / v) sulfanylamide) 100 After mixing to μl, the cells were incubated for 10 minutes at room temperature, and then the absorbance was measured at 540 nm using a microplate reader. Fresh medium was used as untreated group in all experiments. The amount of nitric oxide in the sample was calculated from the sodium nitrite standard curve prepared in the medium.
Example 3. Cytotoxicity Test of Yunpan Herb Extract
Cytotoxicity experiments of yunpanmul extract were determined by MTT assay. Cytotoxicity test cultures were added 5 μl of MTT solution (10 mg / ml in phosphate-buffered saline, pH 7.4) 3 hours before the end of the culture, and the cells were incubated until the end of the reaction. The culture was stopped by adding DMSO to increase the solubility of formazan. Optical density (OD) at 540 nm was measured using an ELISA microplate reader.
Example 4 Measurement of mRNA Production or Expression of iNOS, COX-2, TNF-α and IL-6
Yunpan Sprout Extract was prepared using DMSO to make 100 mg / ml, diluted to concentration, and treated in RAW 264.7 cells, treated with LPS, incubated for 24 hours, and then completely separating RNA from the cells and primers for each gene. RT-PCR was performed.
Example 5 Determination of Protein Production or Expression of iNOS
To determine the effect of yunpanmul extract on the protein expression of iNOS, the yunpanmul extract was treated in RAW 264.7 murine macrophages by concentration and treated with LPS (1 ㎍ / mL) and grown for 24 hours. The total proteins collected by Lysis were electrophoresed and transferred, and then measured using Western blot with iNOS antibody.
Example 6 Measurement of Phosphorylation and Degradation of IκB-α
To determine the effect of yunpanmul extract on the phosphorylation and degradation of IκB-α, the yunpanmul extract was treated by concentration in RAW 264.7 murine macrophages and treated with LPS (1 ㎍ / mL) for 2 hours. After lysing, the cells were lysed and total proteins collected were subjected to electrophoresis and transfer, followed by Western blot with p-IκB-α and IκB-α antibodies.
Example 7.Measurement of Nuclear Migration of P65 Protein
To determine the effect of yunpanmul extract on the p65 protein that migrates into the nucleus, RAW264.7 murine macrophages were treated with yunpanmul extract at different concentrations and simultaneously treated with LPS (1 ㎍ / mL) and grown for 2 hours. The cells were then lysed (lysed), and nuclear proteins (nuclear proteins) collected were electrophoresed and transferred, and then measured using Western blot with p65 and PCNA antibodies.
Example 8 Determination of Activity Binding to DNA in the Nucleus of NF-κB Proteins
To measure the binding activity of NF-κB protein binding to DNA in the nucleus, the yunpanmul extract was treated with concentration of yunpanmul extract in RAW 264.7 murine macrophages and treated with LPS (1 μg / ml) simultaneously. After raising for 2 hours cells were lysed to prepare a nuclear extract. Nuclear extracts (5 μg) were mixed with double-stranded NF-κB oligonucleotides and labeled end with [γ- 32 P] dATP.
The binding reaction was conducted for 30 minutes at room temperature with 50 mM Tris-HCl, pH 7.9, 0.42 M KCl, 6 mM DTT, 0.1 mM EDTA, 5
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101693632B1 (en) | 2016-03-08 | 2017-01-06 | 나천수 | Composition for promoting hair growth comprising extract of Disporum sessile as an effective component |
| KR101956337B1 (en) * | 2017-11-22 | 2019-04-11 | (주)세빛 | Dual color type led module and bar type led lighting apparatus having the same |
| CN110950972A (en) * | 2019-12-13 | 2020-04-03 | 贵州大学 | Polysaccharide extracted by ultrasonic-assisted enzyme method and application thereof |
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2010
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101693632B1 (en) | 2016-03-08 | 2017-01-06 | 나천수 | Composition for promoting hair growth comprising extract of Disporum sessile as an effective component |
| KR101956337B1 (en) * | 2017-11-22 | 2019-04-11 | (주)세빛 | Dual color type led module and bar type led lighting apparatus having the same |
| CN110950972A (en) * | 2019-12-13 | 2020-04-03 | 贵州大学 | Polysaccharide extracted by ultrasonic-assisted enzyme method and application thereof |
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