KR20130059532A - Neutraceutical composition comprising extract of scuscuta japonica chois showing an inhibitory effect of osteoporosis - Google Patents
Neutraceutical composition comprising extract of scuscuta japonica chois showing an inhibitory effect of osteoporosis Download PDFInfo
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- KR20130059532A KR20130059532A KR1020110125560A KR20110125560A KR20130059532A KR 20130059532 A KR20130059532 A KR 20130059532A KR 1020110125560 A KR1020110125560 A KR 1020110125560A KR 20110125560 A KR20110125560 A KR 20110125560A KR 20130059532 A KR20130059532 A KR 20130059532A
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- extract
- osteoporosis
- inhibitory effect
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- disease
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- 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
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/43—Cuscutaceae (Dodder family), e.g. Cuscuta epithymum or greater dodder
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- A23V2250/00—Food ingredients
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Abstract
Description
The present invention relates to a health food composition having an osteoporosis inhibitory effect containing the earth and sand extract as an active ingredient.
Kumamoto H, Ooya K. ExpreASion of parathyroid hormone-related protein (PTHrP), osteoclast differentiation factor (ODF) / receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoclastogenesis inhibitory factor (OCIF) / osteoprotegerin (OPG) in ameloblastomas. J Oral Pathol Med. 2004; 33 (1): 46-52.
2. Reddy SV. Regulatory mechanisms operative in osteoclasts. Crit Rev Eukaryot Gene Expr. 2004; 14 (4): 255-70.
3.Pang M, Martinez AF, Fernandez I, Balkan W, Troen BR. AP-1 stimulates the cathepsin K promoter in RAW 264.7 cells. Gene. 2007; 403 (1-2): 151-8.
Boyce BF, Yamashita T, Yao Z, Zhang Q, Li F, Xing L. Roles for NF-kappa B and c-Fos in osteoclasts. J Bone Miner Metab. 2005; 23 Suppl: 11-5.
Khosla S. Minireview: the OPG / RANKL / RANK system. Endocrinology. 2001; 142 (12): 5050-5.
6.Han SY, Lee NK, Kim KH, Jang IW, Yim M, Kim JH, Lee WJ, Lee SY. Transcriptional induction of cyclooxygenase-2 in osteoclast precursors is involved in RANKL-induced osteoclastogenesis. Blood. 2005; 106 (4): 1240-5.
7.Yamashita M, Otsuka F, Mukai T, Yamanaka R, Otani H, Matsumoto Y, Nakamura E, Takano M, Sada KE, Makino H. Simvastatin inhibits osteoclast differentiation induced by bone morphogenetic protein-2 and RANKL through regulating MAPK, AKT and Src signaling. Regul Pept. 2010; 162 (1-3): 99-108
Murakami A, Song M, Ohigashi H. Phenethyl isothiocyanate suppresses receptor activator of NF-kappaB ligand (RANKL) -induced osteoclastogenesis by blocking activation of ERK1 / 2 and p38 MAPK in RAW264.7 macrophages. Biofactors. 2007; 30 (1): 1-11.
9.H Choi HJ, Park YR, Nepal M, Choi BY, Cho NP, Choi SH, Heo SR, Kim HS, Yang MS, Soh Y. Inhibition of osteoclastogenic differentiation by Ikarisoside A in RAW 264.7 cells via JNK and NF-kappaB signaling pathways. Eur J Pharmacol. 2010; 636 (1-3): 28-35
10. Professor, National College of Oriental Medicine. Herbology, Younglim Publishing Co., Seoul, 1994, pp568-9
Ye M, Li Y, Yan Y, Liu H, Ji X, Determination of flavonoids in Semen Cuscutae by RP-HPLC., J. Pharm. Biomed. Anal. 2002; 28 (3-4): 621-8.
12. Yen FL, Wu TH, Li CC, Lin TJ., Hepatoprotective and antioxidant effects of Cuscuta chinensis against acetaminophen-induced hepatotoxicity in rats., J Ethnopharmacol. 2007; 111 (1): 123-8.
13. Jae Bok Lee, Byung Ryeol Lee, A Study on the Production and Effects of Bacillus spp., 大 韓 鍼灸 學會 誌, 2001; 18: .94-104
14. Koo BS, Choi EG, Park JB, Cho CH, Chung KH, Kim CH., Neuroprotective effect of Chuk-Me-Sun-Dan on NMDA- and AMPA-evoked nitric oxide synthase activity in mouse brain., Immunopharmacol Immunotoxicol. 2005; 27 (3): 499-514.
15. Oh H, Kang DG, Lee S, Lee HS., Angiotensin converting enzyme inhibitors from Cuscuta japonica Choisy., J Ethnopharmacol. 2002; 83 (1-2): 105-8.
Bone metabolism is composed of osteoblasts responsible for bone formation and osteoclasts responsible for bone resorption, and the function of these cells is balanced to maintain homeostasis. However, when osteoblast activity decreases or osteoclast activity increases, bone density decreases and osteoporosis may be induced. Factors causing osteoporosis include menopause, hyperthyroidism, diabetes, stress, lack of smoking and exercise, physical aging, and the use of glucocorticoid drugs in women (1-3). Most of them show osteoporosis by inhibiting osteoblast function or increasing osteoclast activity. Therefore, inhibiting osteoclast function can treat or prevent osteoporosis.
Osteoclasts are differentiated by various differentiation-causing factors in macrophage progenitor cells (4-5). In particular, RANKL (receptor activator of nuclear factor kappa B ligand) secreted from osteoblasts binds to osteoclast precursor cells and RANK (receptor activator of nuclear factor kappa B) present on the surface of osteoclasts. Causes differentiation and activation (6). Differentiated osteoclasts were activated by NF-B, c-Fos, c-jun, AP-1, NFATc1, MAPK, ERK, JNK, p38 activation process, MITF, Src, Akt activation, etc. ) Induces bone resorption by promoting the expression of osteoclast specific proteins such as cathepsin K and calcitonin receptor (7-9).
絲 子 (Cuscuta japonica Chois.) Is a mature ginseng of young ginseng, which has a mild temperament, has the effect of 明目, 肝, 目, 止瀉, and has the effect of 補 肝腎, 益 精髓.方, 遺精, 消渴, 腰痛, 頭眩, 尿 有餘 瀝 used to treat (10). Flavonoids quercetin, its derivatives, and kaempferol are known to be the main constituents of Tosa (11). In recent studies, hepatoprotective effects (12), anti-cancer effects (13), and immunity related to T-cell integrating metabolism have been reported. Enhancement effect (14), antihypertensive effect (15) by the inhibition of angiotensin-converting enzyme (ACE) has been reported, there is no study related to osteoporosis.
The present inventors have completed the present invention by confirming the osteoporosis inhibitory effect by inhibiting the gene expression of the osteoclast differentiation inhibitory effect, osteoclast differentiation and osteoclast activity related factors, such as cathepsin K, TNFa, IL-6, etc. Was done.
In order to achieve the above object, the present invention provides a health functional food for improving and preventing bone metabolism disease containing as an active ingredient tosa extract showing the osteoclast inhibitory effect.
The bone metabolic disease as defined herein includes osteoprosis, paget disease, periodontal disease, metastatic cancer or rheumatoid arthiritis, preferably osteoporosis. Characterized in that.
The earth and sand extract of the present invention is extractable from the earth and sand.
For example, the extract of the present invention, after drying the earth and sand, about 1 to 100 times the weight of the sample, preferably about 1 to 50 times (w / v) volume of water, C 1 to C 4 lower Alcohol or a mixed solvent thereof, preferably water or water and ethanol as an extraction solvent, about 2 to 7 days at a reaction temperature of about 10 to 120, preferably 30 to 90, preferably 12 hours to A second step of repeated extraction 1 to 10 times, preferably 1 to 5 times by a conventional extraction method such as heat extraction, ultrasonic extraction, reflux extraction, ultrahigh pressure extraction for 26 hours, preferably reflux extraction; A third step of filtering and extracting the extract obtained in the above step under reduced pressure; The earth and sand extract of the present invention can be obtained through the step comprising the manufacturing method of the fourth step of freeze-drying the concentrated extract.
In another aspect, the present invention provides a health functional food for the improvement and prevention of bone metabolic diseases containing the production method and tosa extract prepared by the production method.
Tosa extract according to the present invention exhibits the inhibitory effect of osteoclast differentiation, osteoclast differentiation and proliferation-related factors, such as cathepsin K, TNFa, IL-6 gene expression inhibitory effect, it was confirmed that it is useful as a health functional food for osteoporosis inhibition .
The extract of the present invention comprises 0.01 to 99% by weight of the herbal extract based on the total weight of the extract.
However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.
The composition containing the extract of the present invention can be used in various ways, such as drugs, foods and beverages for the prevention and improvement of growth growth promoting action or osteoporosis. Examples of foods to which the present Tosa extract may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. .
&Quot; Health functional food "as defined herein means food prepared and processed using raw materials or ingredients having functionality useful to the human body in accordance with Law No. 6727 on Health Functional Foods." Functional " Structure and function of the nutrient to control or physiological effects, such as to obtain a beneficial effect for health is intended to eat.
The dietary supplement of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition.
In addition, the present invention can be manufactured and processed into a health functional food in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of preventing and improving the target disease.
Health functional foods containing the extract of the present invention can be used in a variety of drugs, foods and beverages for the prevention and improvement of the target disease. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, vitamin complexes, health supplements and the like, and they can be used as powders, granules, tablets, capsules or beverages have.
Foods to which the extract of the present invention can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and health functional foods.
In addition, the present invention can be added to food or beverages for the purpose of preventing and improving the target disease. At this time, the amount of the extract in the food or drink may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml .
The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
In addition to the above, the extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the samples of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the sample of the present invention.
Tosa extract according to the present invention exhibits the effect of inhibiting osteoclast differentiation (TRAP (+) multinucleated cell production), gene expression inhibitory factors such as Cathepsin K, TNFa, IL-6, which are related to osteoclast differentiation and proliferation, osteoporosis It can be usefully used as a dietary supplement for suppression.
1 is a diagram showing the effect on the formation of TRAP (+) multinucleated cells from RANKL treated RAW264.7 cells according to the concentration of Tosa extract (CJ),
2 is a diagram showing the effect on the expression of Cathepsin K according to the concentration of Tosa extract (CJ),
Figure 3 is a diagram showing the effect on TNFa expression according to the concentration of Tosa extract (CJ),
Figure 4 is a diagram showing the effect on IL-6 expression according to the concentration of Tosa extract (CJ).
Hereinafter, the present invention will be described in detail with reference to the following examples and experimental examples.
However, the following examples and experimental examples are illustrative of the present invention, and the content of the present invention is not limited by the following examples and experimental examples.
Example 1 Preparation of Tosa Extract
Tosa was added to 30
Experimental Example 1. Cell Culture and Drug Treatment
The RAW 264.7 cells used in the experiment were cultured in a CO2 cell incubator using DMEM (Dulbecco's modified eagle medium) / 10% fetal bovine serum (FBS) / PC-SM medium, and the number of cells was 96 well plate at 5x10 3 cells / well. And cultured using. After culturing for 24 hours, the culture medium was discarded, and the cells were cultured by exchanging with a-MEM to which 10% FBS, 50 ng / ml RANKL, and 1 ng / ml TGFb were added. Different concentrations of AS were added to the culture. It was incubated for 6 days while changing to the same medium every two days. The experimental group was (1) RANKL-treated control group (N) (2) RANKL-induced control group (C), (3) RANKL-induced group + group administered the 50 ug / ml Tosa extract of Example 1 (CJ50), (4) RANKL induction group + group administered with 100ug / ml earthenware extract of Example 1 (CJ100), (5) RANKL induction group + group administered with 200ug / ml Tosa extract of Example 1 (CJ200).
Experimental Example 2. Measurement of osteoclast generation capacity
After inducing RAW 264.7 cells to osteoclasts with RANKL (receptor activator for nuclear factor B ligand), TRAP-positive multinucleated cells (TRAP (+) MNCs) were identified by staining TRAP, a marker for expression of mature osteoclasts. . Differentiated cells were washed twice with PBS, fixed with 3.7% formaldehyde-citrate-acetone solution for 10 minutes and washed twice with distilled water. Treat the cells prepared by mixing 2% TRAP fast garnet GBC base solution with NaNO3 solution in the same ratio and a solution containing 5% naphtha AS-BI phosphoric acid, 4% acetic acid and 2% tartaric acid. It was left for more than a minute. Observed by light microscopy, TRAP-positive multinucleated cells (TRAP (+) MNCs) having three or more nuclei were counted and used as an index for generating osteoclasts (15).
Experimental Example 3. Effect on Gene Expression
In order to determine the effect on the gene expression of the osteoclast differentiation and proliferation-related factors, such as Cathepsin K, TNFa, IL-6 of the Tosa extract of Example 1 was performed as follows.
3-1. gun RNA detach
Total RNA was isolated from TRAP (+) multinuclear cells differentiated into osteoclasts by treatment with 1 ml Trizol solution (Invitrogen, USA). 100 phenol and 100 chloroform: isoamyl alcohol (24: 1) are added to the isolated RNA, and the mixture is mixed well and centrifuged twice to separate the supernatant. RNA is precipitated using 0.5 ml isopropyl alcohol, washed with 70% ethanol and dried naturally. RNA was thawed in RNAase free water (RNA) free water (Promega, USA), followed by the addition of ALNAISE-free DNase (RNase-free DNase, Promega, USA) and stored at -70 ° C.
3-2. cDNA Produce
Oligo dT 1 was added to the total RNA solution (containing 13 ug RNA) of each of the control and experimental groups isolated in Experimental Example 3-1, mixed carefully, and then incubated at 70 ° C. for 5 minutes. Leave primers at room temperature for about 10 minutes to release the primers, then add cyscript buffer, 0.1 M DTT, dUTP nucleotides, dUTP seeded-labeled nucleotides, Cyscript reverse transcriptase, and then add Mix carefully. Thereafter, the cells were incubated at 42 ° C. for 90 minutes and then left on ice. 2.5 M sodium hydroxide was added thereto, followed by incubation at 37 ° C. for 15 minutes, and neutralized by adding 2 M HEPES buffer to prepare cDNA.
3-3. Real time RT - PCR
First, 5 ug of RNA, 50 ng / random hexamer (random hexamer) 3, 10 mM dNTP 1, were added to the test tube, and DEPC-H 2 O was added to make an RNA / primer mixture of 10. The experimental sample was incubated at 65 ° C. for 5 minutes and then left on ice for at least 1 minute. The reaction mixture was prepared by mixing 10-fold RT buffer 2, 25 mM magnesium chloride 4, 0.1 M DTT 2, RNAase (Promega, USA) 1. The reaction mixture was added to the RNA / primer mixture, left to stand at room temperature for 2 minutes, then SuperScript II RT (Promega, USA) 1 (50 units) was added and incubated at 25 ° C. for 10 minutes. Incubated at 42 o C for 50 min, then inactivated by heating at 70 o C for 15 min and cooled on ice. RNase (Promega, USA) 1 was added and incubated again at 37 o C for 20 min and then stored at -20 o C until use. Add 2x Cyber Green Mix (SYBR Green Mix, Takara, Japan) 12.5, cDNA 0.2, 5 pmol / primer pair mix 1, 11.3 H 2 O to each optical tube (Gibco, USA), 50 o Amplification with C 2 min 1 cycle, 95 °
Treatment of receptor activator for nuclear factor B ligand (RANKL) in RAW264.7 cells increased the expression of TRAP (+) multinucleated cells (MNCs), which promoted osteoclast differentiation. At concentrations of ug / ml, it was shown to inhibit the receptor activator for nuclear factor B ligand (RANKL) -induced osteoclast differentiation (FIG. 1). Cathepsin K, TNFa, and IL-6 were increased in RANKL treatment, and the extracts of sediment were significantly inhibited at 50 ug / ml, 100 ug / ml, and 200 ug / ml. 2, 3 and 4). Therefore, Tosa extract was found to have an excellent effect in inhibiting the expression of Cathepsin K, TNFa, IL-6 gene.
Statistical treatments were compared individually using Student's t-test and determined that there was a significant difference when the p-value was less than 0.05.
Hereinafter, formulation examples of the composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.
Preparation Example 1. Preparation of powder
Tosa extract 200 mg
Lactose 100 mg
The above components are mixed and filled in airtight bags to prepare powders.
Formulation Example 2. Preparation of tablets
Tosa extract 200 mg
Corn starch 100 mg
Lactose 100 mg
Magnesium stearate 2 mg
After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.
Formulation Example 3. Preparation of capsules
Tosa extract 200 mg
Crystalline cellulose 3 mg
Lactose 14.8 mg
Magnesium stearate 0.2 mg
The above components are mixed in accordance with a conventional method for producing a capsule, and filled in a gelatin capsule to prepare a capsule.
Formulation Example 4. Preparation of liquid preparation
Tosa extract 200 mg
10 g of isomerized sugar
5 g of mannitol
Purified water
Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.
Claims (2)
The bone metabolic disease is osteoporosis (osteoprosis), Paget disease (paget disease), periodontal disease (periodontal disease), metastatic cancer (rheumatoid arthiritis) health functional food.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020110125560A KR20130059532A (en) | 2011-11-29 | 2011-11-29 | Neutraceutical composition comprising extract of scuscuta japonica chois showing an inhibitory effect of osteoporosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020110125560A KR20130059532A (en) | 2011-11-29 | 2011-11-29 | Neutraceutical composition comprising extract of scuscuta japonica chois showing an inhibitory effect of osteoporosis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20130059532A true KR20130059532A (en) | 2013-06-07 |
Family
ID=48858330
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020110125560A KR20130059532A (en) | 2011-11-29 | 2011-11-29 | Neutraceutical composition comprising extract of scuscuta japonica chois showing an inhibitory effect of osteoporosis |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20130059532A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019004626A1 (en) * | 2017-06-30 | 2019-01-03 | 동국대학교 경주캠퍼스 산학협력단 | Composition for preventing or treating menopausal syndrome, containing medicinal herb extract as active ingredient |
-
2011
- 2011-11-29 KR KR1020110125560A patent/KR20130059532A/en not_active Application Discontinuation
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019004626A1 (en) * | 2017-06-30 | 2019-01-03 | 동국대학교 경주캠퍼스 산학협력단 | Composition for preventing or treating menopausal syndrome, containing medicinal herb extract as active ingredient |
| US11690888B2 (en) | 2017-06-30 | 2023-07-04 | Dongguk University Wise Campus Industry-Academy Cooperation Foundation | Composition for preventing or treating menopausal syndrome, containing medicinal herb extract as active ingredient |
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