KR101559937B1 - Food composition with safflower seed extract, boswellia extract, Scutellaria baicalensis extract for promoting bone growth and increasing bone density - Google Patents

Abstract

The present invention relates to a food composition for promoting bone growth and increasing bone density, containing a composite as an active ingredient, wherein the composite comprises a safflower seed extract, a boswellia extract, and a Scutellaria baicalensis extract. The food composition containing a composite of natural extracts is nontoxic, is harmless to humans, and has few side effects. In addition, the food composition has effects of increasing osteoblasts, promoting activation and differentiation of osteoblast, and increasing the expression of IGF-1 gene and BMP-2 gene, thereby contributing to children′s growth.

Description

[0001] The present invention relates to a food composition for promoting growth of bone length and for increasing bone density, comprising boswellia extract, boswellia extract and golden extract as an active ingredient. [0002] Food composition containing boswellia extract, boswellia extract, scutellaria baicalensis extract,

The present invention relates to a food composition for promoting growth of bone length and for increasing bone mineral density, and more particularly, to a composition for promoting bone length growth and a bone mineral density containing an ingredient of safflower seed extract, boswellia extract and gold extract as an active ingredient ≪ / RTI >

As the interest in the appearance has increased, it has become a time when the big tiger becomes competitive. This social atmosphere is expressed by the stress on the child and the interest on the growth of the child. However, although many growing children want to grow up to 180 cm for men and 165 cm for women, the average height of Korean men and women is 174 cm for men and 161 cm for women. In addition, westernized eating habits of modern society increased instant foods, refined processed foods, and nutritional imbalance such as loss of micronutrients during processed foods. In addition, excessive use of chemical seasoning, coloring agents, and preservatives may distort homeostasis in the body, resulting in the inhibition of key growth of infants or young children, or the promotion of early development of adult diseases.

Growth, on the other hand, refers to an increase in the number of cells, an increase in skeletal and muscular strength, and an increase in the anatomical and morphological size and function of not only the kidney but also the organs of the body. This growth is made by the action of hormones. The growth hormone produced in the pituitary gland mainly acts on the liver but also acts on the growth plate, gonad and adipose tissue of the bone and induces the production of insulin-like growth factor-1 (IGF-1) do. Growth hormone acts on target cells to induce IGF-1 production and cell differentiation, after which IGF-1 induces cell proliferation (Waters MJ et al., Clin Exp Pharmacol Physiol. 1999. 26: 760-764). Therefore, the physiological activity of growth hormone is known to be related to IGF-1 in many parts.

Generally, it is believed that the genetic influence is the largest in human height growth, but the genetic effect is 23% and the remaining 77% is determined by the acquired effect. Key growth by acquired effects is due to the combination of proper nutritional supply and body hormones. If any of these external or internal mechanisms of action occurs, normal growth can not be expected. Therefore, in order for normal growth to occur, it is important to ensure normal hormone secretion and adequate nutrition.

On the other hand, growth hormone therapy is used as a treatment for growth hormone deficiency such as pediatric growth hormone deficiency syndrome, Turner syndrome, short stature due to chronic renal failure, Frieder-Willie syndrome, Noonan syndrome, Method. In modern medicine, if the growth is within the third percentile in the same age and sex group, diagnosis is made as short stature and gonadotropin is administered. However, the hormone therapy used for low-stature can lead to various side effects such as leukemia, central nervous system tumor, hypothyroidism, epilepsy, hyperglycemia and diabetes (Yang, JS, And the economic inefficiency in the growth effect compared to the high cost. In addition, it can give psychological burden to a child who should be injected almost every day in 5 ~ 7 weeks, and as the treatment period becomes longer, the growth effect is not good and the capacity of the growth hormone used as a high dose should be increased continuously. And the ethical problem of using therapeutic drugs.

In order to replace the hormone therapy for the growth of the child's key which has raised such problems, various health supplement foods which can promote the growth of the key by supporting the nutritional balance have been developed, but merely a combination of various ingredients But the effect is insufficient, and only the growth of the skeleton is over-emphasized, so that side effects which can abnormally activate the body metabolism of the recipient are not taken into consideration and practically not utilized.

Korean Patent Laid-Open No. 10-2009-0011654 (Date of Publication: 2009.02.02) describes a technique for a growth promoting food composition containing a herbal extract of Hwanggi, Baekjoo, Baekbokryeong, Dhamma, Sine, Ginseng and licorice have. Korean Patent No. 10-1376052 (Registration Date: Mar. 13, 2013) describes a method for promoting bone growth, which contains bokbunja extract obtained by adding 70% (v / v) ethanol to bokbunja and heating and refluxing for 8 hours. Techniques for food compositions are described. Korean Patent No. 10-0890991 (registered on March 23, 2009) discloses a herbal composition comprising a complex herbal extract comprising 2 parts by weight of a herbicide extracted with a lower alcohol of C1 to C4 or an aqueous solution thereof and 1 part by weight of a herb extract as an active ingredient, A description of a growth promoting health supplement is provided.

It is an object of the present invention to develop and provide a food composition containing a complex composed of safflower seed extract, boswellia extract and golden extract as an active ingredient, and having an effect of promoting bone length growth and an effect of increasing bone density.

In order to attain the above object, the present invention provides a composition comprising, as an active ingredient, a complex composed of 30 to 65% by weight of safflower seed extract, 10 to 40% by weight of boswellia extract and 10 to 55% Provided is a food composition for promoting length growth and for increasing bone density. The complex thus formed exerts excellent osteoblast proliferation effect, osteoblast differentiation and activity enhancement effect, and IGF-1 and BMP-2 gene expression enhancement effect. In addition, the above complex composition exhibits more excellent activity than the single extract, safflower seed extract, boswellia extract and gold extract.

Osteoblasts originate from mesenchymal stem cells. Mineralization, such as calcium formation by osteoblast differentiation, not only maintains bone strength but also plays an important role in the homeostasis of calcium and hormone metabolism throughout the body. Calcium formation by osteoblast differentiation is regulated by vitamin D and parathyroid hormone. Bone formation by osteoblast differentiation is induced by bone morphogenetic protein (BMP), Wnt ), MAP kinase, calcineurin-calmodulin kinase, NF-κB (nuclear factor kappa B), and activator protein 1 (AP-1) Alkaline phosphatase (ALP) associated with osteoblast differentiation is synthesized in the early differentiation stage by the interaction of osteoblast, osteocalcin, type I collagen, etc. associated with mineralization . The promoter for activating such osteoblast cells is generally required to be administered to a patient for a long period of time. Therefore, it is desirable that the oral administration is possible with less toxicity, and development of functional food for promoting osteoclast activation without toxicity is required. Accordingly, the present inventors have studied the novel food raw materials exhibiting the osteoblast activity promoting effect from the natural substances having no toxicity and side effects. As a result, the optimum blending ratio of the safflower seed extract, the boswellia extract and the golden extract was obtained, and the safflower seed extract To 65% by weight of boswellia extract, 10 to 40% by weight of boswellia extract and 10 to 55% by weight of gold extract were excellent in bone length growth promoting effect and bone mineral density increasing effect.

On the other hand, safflower is a perennial plant belonging to Asteraceae, which is called safflower, and has been cultivated primarily for medicinal purposes in Korea, Japan, and China. The safflower seeds are rich in vitamins (vitamin E, B1, and B2), rich in minerals (K, P, Ca, and Mg), and also have 15 to 30 percent lipid. As for their fatty acid composition, palmitic acid Palmitoeic acid and stearic acid and unsaturated fatty acids such as oleic acid, linoleic acid and linolenic acid and unsaturated fatty acid content of 83 to 87 %. In particular, it contains 69 to 79% of linoleic acid, which is known to be effective in lowering cholesterol concentration, which causes hypertension and arteriosclerosis.

Boswellia is a granular resin extracted from twigs, one of the tallest shrubs in the Indian highlands. It is used for medicines in diabetes, skin diseases, fever, neurological diseases, rheumatism and diarrhea in India. It is now recognized as a non-steroidal anti-inflammatory, functional food. The major components of boswellia are β-boswellic acid, AKBA, terpenoid, and β-boswellic acid is known to inhibit cartilage cell necrosis, promote chondrocyte proliferation, And the like.

Golden ( Scutellaria baicalensis ) is the root of a perennial herbaceous plant, a perennial herbaceous plant belonging to the dicotyledonous plant Lamiaceae. The toxicity is low and 30 kinds of flavonoid compounds are separated as the main component. Such gold pharmacological actions include antipyretic, diuretic, antiviral, antimicrobial, sedative, hypotensive, hyperglycemic, intestinal, intestinal (exercise) (anaphylaxis) action, anti-inflammatory action, anti-allergic action, antioxidative action, and cell immune stimulating action, and the herbal medicine is used as a prescription medicine for various herbal medicine based on these actions.

On the other hand, the safflower seed extract or the boswellia extract or the gold extract is preferably extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixture thereof as an extraction solvent.

The safflower seed extract or the boswellia extract or the gold extract may be extracted using hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.

The extract may be in the form of a powder obtained by lyophilization or hot-air drying, preferably comprising a concentrate obtained by extraction, filtration and concentration under reduced pressure or vacuum concentration.

Further, it is preferable that the above-mentioned composite is contained in an amount of 0.01 to 50% by weight of the entire food composition. If the amount is less than 0.01% by weight, the effect of increasing the bone length growth and increasing the bone density is insignificant. If the amount is more than 50% by weight, the increase in the effect on the usage amount is insufficient.

Meanwhile, the bone-length growth promoting and bone mineral density-enhancing food composition of the present invention is preferably used for promoting growth of bone length and for increasing bone density, A dietary supplement, a dietary supplement, a dietary supplement, and other health supplement foods, but the present invention is not limited thereto.

The bone-length growth promoting and bone mineral density enhancing food composition of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of foods. The carrier, the negative active material and the diluent are, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinyltolydone, water, honey, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral may be unique. When formulated, it is prepared using diluents or excipients such as fillers, weights, binders, humectants, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

The bone-length growth promoting and bone mineral density-enhancing food composition of the present invention can be used for various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers (cheese, Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. It can also contain natural fruit juice and pulp for the production of fruit drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additive is not so important, but is generally selected in the range of 0 to 50 parts by weight relative to 100 parts by weight of the composite of the present invention.

According to the present invention, it is possible to provide a food composition containing a complex composed of safflower seed extract, boswellia extract, and golden extract as an active ingredient and having an effect of promoting bone length growth and an effect of increasing bone density.

In addition, the present invention can provide a food composition for promoting growth of bone length and for increasing bone density, which comprises a complex composed of a natural extract having no toxicity, no side effects, and being harmless to human body as an active ingredient.

In addition, the bone-length growth promoting and bone mineral density enhancing food composition of the present invention is excellent in osteoblast-promoting effect, osteoblast activation and differentiation promoting effect, and IGF-1 gene and BMP-2 gene expression increasing effect, It can help promote growth.

Fig. 1 is a graph showing an experiment result of confirming the effect of the complex on osteoblast proliferation.
FIG. 2 is a graph showing an experimental result of measuring the activity of basic phosphatase (ALP) in order to confirm the effect of the complex on the osteoblast activity.
FIG. 3 is a graph showing the experimental results of measuring the degree of calcification by alizarin staining in order to confirm the effect of the complex on the differentiation of osteoblasts.
FIG. 4 is a graph showing an experiment result of confirming the effect of the complex on IGF-1 gene expression.
FIG. 5 is a graph showing an experiment result of confirming the effect of the complex on BMP-1 gene expression.

Hereinafter, the structure of the present invention will be described in detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Preparation Example 1: Preparation of safflower seed extract]

After drying 1 kg of safflower seeds, 300 g of the powder obtained by grinding was put in a house collector, and 3 liters of water was added and extracted repeatedly three times at 90 ° C for 6 hours to obtain a safflower seed extract. Thereafter, the mixture was filtered under reduced pressure through a filter paper (whatman no. 1, England), concentrated under reduced pressure at 60 ° C in a vacuum rotary condenser, and then dried with hot air to prepare safflower seed extract powder.

[Preparation Example 2: Preparation of boswellia extract]

300 g of boswellia gum resin was placed in a bowl and then 1.8 liters of 95% alcohol was added and extracted three times at 70 DEG C for 6 hours to obtain a boswellia extract. Thereafter, the mixture was filtered under reduced pressure through a filter paper (whatman no. 1, England), concentrated under reduced pressure at 60 ° C in a vacuum rotary condenser, and then dried with hot air to prepare boswellia extract powder.

[Preparation Example 3: Preparation of golden extract]

After drying 1 kg of gold, 300 g of the powder obtained by grinding was put in a bowl and 3 ℓ of water was added, and the mixture was repeatedly extracted three times at 90 ° C for 6 hours to obtain a golden extract. Thereafter, the mixture was filtered under reduced pressure through a filter paper (whatman no. 1, England), concentrated under reduced pressure at 60 ° C in a vacuum rotary condenser, and then dried with hot air to prepare a gold extract powder.

[Examples 1 to 4: Preparation of composite material]

The complexes were prepared by mixing the safflower seed extract powder, boswellia extract powder and gold extract powder prepared in Preparation Examples 1 to 3 at different weight ratios as shown in Table 1 below.

Safflower seed extract
(weight%) Bosswellia extract
(weight%) Golden extract
(weight%) Example 1 58.3 25 16.7 Example 2 50 33.3 16.7 Example 3 50 16.7 33.3 Example 4 33.3 16.7 50

[Experimental Example 1: Confirmation of effect of composite on osteoclast survival rate]

In this experimental example The effect of the single extract prepared in Preparation Example 1 and the composite prepared in Examples 1 to 4 on osteoclast survival rate was examined.

(1) Osteoclast culture

The osteoblast MG-63 cell line was purchased from the American Tissue Culture Collection (ATCC) and used for the experiment. MG-63 cells were treated with 1% penicillin-streptomycin (Hyclone Laboratories, Logan, Utah, USA) in 10% FBS (Hyclone Laboratories, Logan, Utah, USA) and MEM (minimal essential medium) 1% L- glutamine (L-glutamine, Hyclone Laboratories, Logan, Utah, USA), 0.1% gentamycin (gentamycin, Gibco, Grand Island, NY, USA) was added into the flask to 75T 37 ℃, 5% CO ₂ . ≪ / RTI > After that, the cells were changed every 2 days and used for 4 to 5 days.

(2) Confirmation of effect of composite on osteoclast survival rate

MG-63 cells were placed in a 96-well plate at a concentration of 1 × 10 ⁴ cells / well in a MEM culture medium supplemented with 10% FBS and stabilized for 12 hours. 3, Examples 1 to 4) were dissolved in 0.2% DMSO and treated at a concentration of 5 μg / ml and cultured for 48 hours. After incubation, each well was treated with 20 μl of MTT (3- (4,5-methylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) reagent and incubated for up to 3 hours at 37 ° C. in an incubator After removing the medium and MTT reagent, 200 μl of DMSO was added and the absorbance was measured at 560 nm using an ELISA reader (VERSAMAXSL-20 Molecular Devices, Seoul, Korea). The control group was not treated with the sample, and the results are shown in Table 2 below.

Cell survival rate (%) Control group 100 ± 10.4 Example 1 118.4 ± 12.1 Example 2 162.9 ± 8.1 Example 3 130.5 ± 5.0 Example 4 108.4 ± 12.9 Production Example 1 117.5 ± 7.1 Production Example 2 110.5 ± 9.2 Production Example 3 102.7 13.3

As a result, it was confirmed that osteoblast survival rate was higher than that of single extract (safflower seed extract, boswellia extract, and gold extract). In addition, it was confirmed that Example 2 (50% by weight of safflower seed extract, 33.3% by weight of boswellia extract and 16.7% by weight of gold extract) among the complexes showed the highest osteoclast survival rate. Hereinafter, the experiment was conducted using Example 2 in which the effect was most excellent.

[Experimental Example 2: Confirmation of effect of the complex on osteoblast proliferation]

In this experiment, we investigated the effect of complex on osteoblast proliferation.

MG-63 cells were seeded in a 96-well plate at a concentration of 1 × 10 ⁴ cells / well in a MEM culture medium supplemented with 10% FBS, stabilized for 12 hours, Were dissolved in 0.2% DMSO and treated at 0, 1, 5, 10, 20, 30, and 40 ㎍ / ㎕, respectively, for 48 hours. After incubation, the wells were treated with 20 μl of MTT reagent and incubated for up to 3 hours at 37 ° C in an incubator. After removing the medium and MTT reagent, 200 μl of DMSO was added and the absorbance was measured at 560 nm using an ELISA reader And three measurements were made to determine the mean value and the standard error. The control group was not treated with the complex.

As a result, the MG-63 cell proliferation was increased at all the concentrations of the complex-treated control group. Especially, at all treatment concentrations except 40 ㎍ / ㎕ concentration, it was confirmed that cell proliferation was significantly increased compared with the control group. From the above results, it was confirmed that the complex did not show cytotoxicity, and the osteoblast proliferation effect was excellent (FIG. 1).

[Experimental Example 3: Confirmation of the effect of the compound on the activity of the osteoblast - Measurement of alkaline phosphatase (ALP) activity]

In this experiment, the activity of alkaline phosphatase (ALP) was measured in order to examine the effect of the complex on osteoblast activity.

Osteoblast cells have a basic phosphatase (ALP) in the cell membrane, which is found at high concentrations in the outer membrane of the cells and in the calcified tissues, and regulates the transport, cell division and differentiation during calcification, (Cancedda et al., Int Rev Cytol. 1995. 159: 265-358).

MG-63 cells were placed in a 96-well plate at a concentration of 1 × 10 ⁴ cells / well in a MEM culture medium supplemented with 10% FBS and stabilized for 12 hours. Then, the complex (Example 2) , 5, 10, 20, and 30 ㎍ / ㎕, respectively, and cultured for 48 hours. After incubation for 48 hours, the medium was removed and ALP activity was measured using an ALP measurement kit. The assay was performed using the abcam 'Alkaline Phosphatase Assay Kit (abcam, Massachusetts, USA)' according to the manufacturer 's method and assayed using an ELISA reader.

As a result, the basic phosphatase activity of the control group without complex treatment was found to be 56.31 U / ml, while the activity of the basic phosphatase was increased by 20% . Especially, at 10 ㎍ / ㎕ and 20 ㎍ / ㎕, it was confirmed that it increased more than 25% at 76.88 U / ㎖ and 75.88 U / ㎖. From the above results, it was confirmed that the composite was excellent in promoting osteoclast activation (FIG. 2).

[Experimental Example 4: Determination of the effect of the composite on the differentiation of osteoblasts-Measurement of mineralization by alizarin-red staining]

In this experimental example, the degree of mineralization by alizarin-red staining was measured to examine the effect of the composite on osteoblast differentiation.

The osteoblast differentiation occurs with bone formation following bone growth, and the degree of calcification is an important index for cell differentiation (Goldstein et al., New York: McGraw-Hill, 2001. 2863-2913).

MG-63 cells were added to 6-well plates at a concentration of 5 × 10 ⁵ cells / well in DMEM supplemented with 10% FBS and stabilized for 12 hours. Then, the complex (Example 2) Were treated at 0, 5, 10, 20, and 30 ㎍ / ㎕, respectively, and cultured for 6 days. After incubation, the medium was removed, washed with PBS, fixed with 4% paraformaldehyde solution at room temperature for 15 minutes, and paraformaldehyde solution was removed and washed one more time. After washing, 1 ml of a 2% Alizarin Red S solution was treated and allowed to stand at room temperature for 20 minutes. Thereafter, the cells were washed 3 to 4 times with PBS, and 10 mM sodium phosphate solution (pH = 7.0) containing 10% (w / v) cetylpyridinium chloride was added to each well 1 ml each was added and dissolved for 10 minutes. The degree of staining was measured at 570 nm using an ELISA reader. The control group was treated with the complex-untreated group, and the degree of calcification was calculated based on the absorbance of the control group.

As a result, the degree of calcification was significantly increased in the cells treated with the complexes compared to the control without the complex, and the lime degree was increased by 22.49% and 27.36% at 10 ㎍ / ㎕ and 20 ㎍ / ㎕ respectively . From the results described above, it was confirmed that the treatment of the complex improves the calcification according to osteoblast differentiation (FIG. 3).

Experimental Example 5: Confirmation of effect of complex on IGF-1 (Insulin-like Growth Factor-1) and BMP-2 (Bone morphogenetic protein-2)

In this experiment, real-time PCR (Real-Time PCR) was performed to confirm the effect of the complex on gene expression of IGF-1 (Insulin-like Growth Factor-1) and BMP-2 (Bone morphogenetic protein- Respectively.

Insulin-like Growth Factor-1 (IGF-1) is a growth factor associated with growth hormone and is secreted by the anterior pituitary gland stimulated by growth hormone. IGF-1 is known to affect chondrocyte proliferation, especially in the growth plate (Hallahan et al. Nat Med. 2003. 9: 1033-1038). In addition, it also participates in the expression mechanism of growth hormone, regulating the expression of growth hormone and controlling cell differentiation, proliferation, and maturation in most places (Schiller et al. Bone. 28: 362-369).

Bone morphogenetic protein-2 (BMP-2) plays an important role in bone formation as a growth factor belonging to the TGF-β (superfamily) and transfection growth factor (Stein et al. FASEB J. 1990. 4 : 3111-3123). In particular, it is known that retinoids regulate the substances responsible for the proliferation of chondrocytes (Wang EA et al., Proc Natl Acad Sci USA 1990. 87: 2220-2224), the osteogenic inhibitor noggins (Stein et al. FASEB J. 1990. 4: 3111-3123).

MG-63 cells were added to 6-well plates at a concentration of 5 × 10 ⁵ cells / well in DMEM supplemented with 10% FBS and stabilized for 12 hours. Then, the complex (Example 2) Were cultured for 6 days at 0, 5, 10, 20, and 30 ㎍ / ㎕, respectively. Cells were collected and analyzed according to the manufacturer's method of RNeasy extraction kit (QIAGEN, Germantown, Maryland, USA) RNA was extracted. Then, 4 μl of 5 × iScript select reaction mix was added to 5 μg of total RNA using the iScript Select cDNA Synthesis Kit (BIO-RAD, Singapore, Singapore) for the synthesis of cDNA, and Oligo (dT) Primer set 2 5 μl of RNA sample, and 8 μl of Nuclease-Free Water, and 1 μl of iScript Reverse Transcriptase was added to each well. After incubation at 42 ° C for 60 minutes and at 85 ° C for 5 minutes, the synthesized cDNA was used for the PCR reaction. The real-time PCR was performed in accordance with the method of 'iQ SYBR Green Supermix (BIO-RAD, Singapore, Singapore)' and the 'Step One Real-Time PCR System (Applied Biosystems, Foster, . The final concentration of the mixed solution for PCR was 2 μl (10-100 ng) of cDNA, 10 μl of 2 × iQ SYBR Green supermix, 1 μl of each forward and reverse primer (250 nM), and 7 μl of H ₂ O. PCR was carried out at 95 ° C for 10 minutes, followed by 15 cycles at 95 ° C for 15 seconds, at 58 ° C for 15 seconds, and at 72 ° C for 30 seconds for 40 cycles. Finally, PCR was carried out at 95 ° C for 15 seconds, , Followed by 15 seconds at 95 ° C for PCR analysis. The primers used in the reaction were as shown in Table 3 below.

β-actin
forward 5'-GAC CCA GAT CAT GTT TGA GAC CTT-3 ' reverse 5'-CCA GTA GTA CGG CCA GAG GTG TA-3 ' RUNX2

forward 5'-TGC TTC CAT CAC AGT ACC ATC ATT-3 ' reverse 5'-AAA AAC GAA CCC AAC AGT TCA GA-3 ' IGF-1
forward 5'- TTC CGG AGC TGT GAT CTA AGG-3 ' reverse 5'-CGG ACA GAG CGA GCT GAC TT-3 ' BMP2
forward 5'-TGC TAG TAA CTT TTG GCC ATG ATG-3 ' reverse 5'-GCG TTT CCG CTG TTT GTG T-3 '

IGF-1 expression levels were increased in the cells treated with 20 μg / μl and 30 μg / μl of the complex, respectively, as compared with the control (1.3, 1.5).

As a result of the measurement of the expression level of BMP-2, it was confirmed that the expression of BMP-2 was increased to 1.6 and 1.7 times, respectively, in the cells treated with 20 / / 복 and 30 / / 복 of the compound (Fig. 5).

From the above results, it was confirmed that the composite showed excellent osteoblast differentiation and proliferation effect.

[Example 5: preparation of a food composition for promoting growth of bone length and for increasing bone density]

In this Example, a food composition for promoting growth of bone length and for increasing bone density was prepared as follows.

(1) Manufacturing of wire

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and then the mixture was prepared into powder having a particle size of 60 mesh by a pulverizer. Black beans, black sesame seeds and perilla seeds were each steamed and dried by known methods, and then power distribution and pulverization were carried out to prepare powder having a particle size of 60 mesh. Thereafter, 30 wt% of brown rice, 15 wt% of yulmu, 20 wt% of barley, 9 wt% of glutinous rice, 7 wt% of perilla seed, 8 wt% of black soybean, 7 wt% of black sesame, 3 wt% of complex (Example 2) By weight and 0.5% by weight of sulfur were mixed to prepare an electric wire.

(2) Production of chewing gum

Chewing gum was prepared in a conventional manner by combining 20% by weight of a gum base, 76.9% by weight of sugar, 1% by weight of a flavor, 2% by weight of water and 0.1% by weight of a complex (Example 2).

(3) Candy manufacturing

Candies were prepared in a conventional manner by mixing 60% by weight of sugar, 39.8% by weight of starch syrup, 0.1% by weight of fragrance and 0.1% by weight of the complex (Example 2).

(4) Manufacturing of biscuits

By weight of starch, 0.77% by weight of glucose, 0.78% by weight of glucose, 11.78% by weight of palm shortening, 1.54% by weight of ammonia, 0.17% by weight of sodium bicarbonate, 0.16% by weight of sodium bisulfite 1.45 weight% of rice flour, 0.0001 weight% of vitamin B1, 0.0001 weight% of vitamin B2, 0.04 weight% of milk fractions, 20.6998 weight% of water, 1.16 weight% of whole milk powder, 0.29 weight% 0.29% by weight of spray salt and 7.27% by weight of spray oil and 1% by weight of the composite (Example 2) were mixed to prepare a biscuit by a conventional method.

(5) health drink manufacturing

0.0001 wt.% Of niacinamide, 0.0001 wt.% Of sodium riboflavin hydrochloride, 0.0001 wt.% Of pyridoxine hydrochloride, 0.001 wt.% Of inositol, 0.002 wt.% Of orthoacetic acid, 98.7362 wt. Example 2) were mixed together to prepare a health drink.

(6) Production of sausages

(Example 2) was mixed with 65.18 wt% of pork, 25 wt% of chicken meat, 3.5 wt% of starch, 1.7 wt% of soybean protein, 1.62 wt% of salt, 0.5 wt% of glucose and 1.5 wt% of glycerin, Sausages were prepared.

(7) Health supplement manufacturing

50% by weight of the complex (Example 2), 16% by weight of guar gum enzyme hydrolyzate, 0.01% by weight of vitamin B1 hydrochloride, 0.01% by weight of vitamin B6 hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, And 1.85% by weight of corn starch were blended to prepare a refillable health supplement.

Claims (3)

50% by weight of safflower seed extract, 33.3% by weight of boswellia extract and 16.7% by weight of gold extract; or
50% by weight of safflower seed extract, 16.7% by weight of boswellia extract and 33.3% by weight of golden extract; As an active ingredient, the bone-length growth promoting and bone-density increasing food composition.
The method according to claim 1,
The safflower seed extract or the boswellia extract or the golden extract may contain,
Wherein the extract is extracted with water, a lower alcohol having 1 to 4 carbon atoms or a mixture thereof as an extraction solvent.
The method according to claim 1,
The composite may comprise,
Wherein the food composition comprises 0.01 to 50% by weight of the whole food composition.

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